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1. ani CellCult ny FilterPla ERRE i all CallCult FilterPla Edit Method n Method Bu For Help press Fl Fe 1108 000 i 18 4 This could be a part of a normal plasmid DNA isolation protocol combined with a PCR setup It is very useful to work with parallel processes using the vacuum box During the vacuum procedures the Microlab STAR robot goes on with pipetting and thus saves time Accordingly this section will also explain how to use the parallelism within the Microlab STAR User SW The first step in our program is initialize Then come the following steps lj Hamilton Method Editor Y ual med l x E File Edit wiew Method Tools Window Help lel x O orl fm x meee ee eo em tet roe SetCarrier Temperature on ML_STAR Loop Break Rack type TOC1 Temperature 10 C Mode 0 E ee No wait for temperature Heat up time 60 a E Return value count 3 m FA Sequence Hi Mii Pipette with Tips trom ML_STAR Smart Steps H File Handling c Timer Comm a Dialogs m Communicati W ey Instrument short name ML_STAR Standard pipette procedure Mode Simple 1 1 Pipette volume ul S00 Aspirate sequence ML_STAF CellCuture Dispense Sequence ML_STAR FiterPlateConditionSitte controlling Sequencel ie BE wait For Eve sel evel FE kort Begin parallel process process Brees LE ES l y yj IO ace cen eae err er e
2. sssssecccocsssssececcosssssccccocosssseeceoosssssseecooosssssecescsssssseeesosssssssee 13 Dot APCE A aona O O E 13 Did ASP AC OU e a E stan sneeeecaonteguadaae 13 Deed ISDE MSG eiaeaesialcortane a smote nts a heantaae tise ted ausaainnclnndes a cheney ate atusnieds 16 Zt ARTO fos OT oense op ieee a a adore aenearnst ea stata ee eA 17 23 AVOIGING Contamination diorite a a a aa a esar 17 2 4 Liquid Classes Pipetting Modes and More sssssssecccoossssssecccossssesececoosssssecesoosssssececosssssssee 17 Zeal digud Handins Example Seega Ea a EE 18 2 95 Process Controkosioicjissssnsasi a aa aaa aea aaia aaa 20 2951 Momtoredt Air Displacement resar aaan Ea a a Aa 20 2 5 2 Capacitance Based Clot Detection ercran paa a r Ea EE EEEE EERE 22 3 Description of the Microlab STAR sssssssssssssesoossseecoosseeecocssseecoossseecoossseecoossseecoossseceosssssee 23 XL The Standard Microlab STAR eiei dira aeaea n a aa EER SOS OAA OaE 23 SL OOS a E E E 24 S22 AS 12 Or TO Pipe Cris Channel Seea a a N A Oa 24 322 2 Mo hkand Low V olumie hanne Is Sranan nana E A T E ecmeaktads 25 Se PRULOIOAG ODOM east AEE S AEA 26 3 3 PRECOSSOTICS sauais aa iiaa aa aa a aa a aaa aa a Eea Ea aoaaa aea aat Zi Died SC ANIC IS sates ces ae san A E aa wae ann Gantt A a E E een iataa tenes teat te ude hah na Gaeuthan nantes tae ntues 27 e DIPODE Sansen cn cyseaan sas E E E ctcace se acanan ta teaee sauce Ea 28 Soo Neede Wasi SLAlION sasan nomon ao bedeasesden
3. water Standard olume Water Dispense et T Cancel Help Select Standard Pipette Procedure Simple Mode Tip Sequence ML_STAR Tips Volume 50 ul residual Volume 0 Tip handling After each dispense Aspiration sequence ML_STAR Samples as controlling sequence ML_STAR Plate as dispense sequence Choose Not Reload for the Sample Sequence Choose Reload or Multiple use for the Plate sequence This is of no influence here since the aspiration sequence is controlling and equally long or even shorter by reduction on runtime than the dispense sequence Dispense mode is Jet Liquid is Water LLD settings are capacitance on aspiration sensitivity low submerge depth 2 mm and fixed height 2 mm on dispense as in the example PlateToPlatePipette Page 138 of 225 610766 02 Microlab STAR User Manual Click Finish Check the input on the summary that appears and click OK Now drag the SMART Step Unload to the next line Sequences i 4 ML_STAR Plate 2 ML_STAR Samples ML_STAR Tips 4 Click on add all sequences to unload the complete deck Click OK to add the step Now your method is ready to use What happens if you run this method Within the loading step the sample sequence was chosen to be reducible This means that at runtime the user sees the following dialog enabling reduction of the number of samples from any position Load Sequence Miel E3 Load the th
4. Processing step Defines what where and with what has to happen on the Microlab STAR It is defined in accordance with the methods the loading and the tasks These are the foundation stones of Microlab STAR methods Rack Grouping of containers in a mechanical manipulable unit Rack identification Barcode for Rack identification Random access Means that every channel can access everywhere on the work area Run Execution of the processing steps defined in the relevant method with the aim of processing one or more liquids and containers e g MTP The run is a series of timed commands in order to carry out processing on the Microlab STAR according to the processing plan The run can be interrupted to load more elements Then processing goes on according to a newly calculated processing plan with the run being started again Loading is not a part of the run 610766 02 Page 213 of 225 Microlab STAR User Manual Run abort Cancellation of a run by the user or by the Microlab STAR Run Visualisation Visualization of the present work status of the Microlab STAR Sample Refers to a liquid in an unambiguously identified container which is to be processed Stacker Storage unit for racks Tip Disposable tip for dispensing Tip rack Frame that holds the tips together Tip waste Container into which the tips are dropped Touch off Type of dispensing whereby the tip or needle approaches the bottom of th
5. Deck Z 0 0 0 x Pipetting Autoload i i Channel eni Pipetting Carrier for tubes Waste Head microtiter plates Container etc The Microlab STAR The Microlab STAR s deck is divided into 54 equal tracks T for loading carriers with predetermined positions This obviates the need for precise measurement of positions The deck has partitions for a maximum of 54 specialised 1 T carriers for sample tubes or a maximum of 9 6 T carriers for microtiter plates and CO RE tips An additional partition space is provided for the tip waste container 610766 02 Page 23 of 225 Microlab STAR User Manual The instrument s internal coordinate system is shown in the figure located at its origin The Microlab STAR is equipped with a pipetting arm typically containing 8 pipetting channels which work independently The pipetting arm can move in an X direction whereas each pipetting channel can move relatively independently both in a Y and a Z direction The Microlab STAR supports pipetting with disposable tips or with needles The pipetting channels have a set travelling height of 245 mm above the origin or 145 mm between the tip of the disposable tip and the deck of the instrument That means that when a channel is to move from one location on the deck to another it automatically does so at that particular height This is a safety precaution so that channels will not collide with any items that may be on the deck The instrument
6. Dispense jet Liguid class I s tandardYolume_ Water Dispenselet il Liguid level detection LLO Capacitive LLD sensitivity Pressure LLD sensitivity 0 Off 0 Ott al Submerge depth Fis height from bottom Max height difference 2 fo fo Channel settings Advanced Error settings ome e For sequence choose ml_star Source to aspirate from the source plate Select Automatic as sequence counting For volume enter 50 ul Select Aspiration as the aspiration mode Choose Standard volume tip without filter for the tip class and Dispense jet as dispense mode Then choose an item from the Liquid class dropdown list For questions about these parameters refer Chapter 2 of this manual The Art of Pipetting Select medium as the capacitive LLD sensitivity Type in 1 0 or 1 as additional submerge depth Click Advanced A window appears Aspirate Advanced Liguid following during aspirate and mi 1 On 7 PreninsingMix settings Cycles 0 Ot Mis position Volurrie E A pl Cancel Help Page 130 of 225 610766 02 Microlab STAR User Manual Set liquid following during aspirate and mix to on allowing the channel to follow the falling liquid level during aspiration You may now enter the settings for mixing The mix position is the submerge depth used for mixing moving the tip downwards
7. E Frogrammet HAMILTONS Contigs ML_S TARA Liguids_ ctg Available liquid classes Selected liquid classes Low Yolume yw aterDispenseS uracel 000er Low ol ater ispenses ubace 2 Needle 300A est ol aterD spenselet Needed ol ater pense et Needle sUUy olaterDispenses urbace NeedeS0 ow aterDispenselet NeedeSttyolyaterDispenseS urace Standardyolume DMSO Dispenselet Standarnd olume DMSO DispenseSurface Standard olume EtWH Dispenselet Standard olume Glycerin Dispense et Standand olume_ Glycerin DispenseSurface Standard olume_Serum Aliquotlet Standardy olume Water Aliquot et l Add all standardyolume_ Water Dispense et Standard olume Water DisnenseS urace Remove all OK Cancel Import Liquid s Window Open the desired liquid configuration file using the Open file button Select the required liquids Close the dialog with OK to import all the liquids selected In addition a similar dialog is available for exporting liquid classes to configuration files This dialog is accessed by clicking on Export 610766 02 Page 105 of 225 Microlab STAR User Manual 12 Microlab AVS Automated Vacuum System 12 1 Integration of the AVS The Microlab AVS is a separate software component that comes with the vacuum box It has to be installed separately For convenience you may wish to add the vacuum box permanently to the deck layout The following example describes the programming of the Automated Vacuu
8. The method we are now going to describe does exactly the same as the method OnePlateToPlatePipette described in section 13 2 The only difference is that the method is now written using single steps to illustrate the difference No sample reduction is possible here First create the deck layout in the same way as described for the method OnePlateToPlatePipette This time save it under the name OnePlateToPlate lay Click on Edit Method Link the deck layout OnePlateToPlate lay to your new method as described above Page 124 of 225 610766 02 Microlab STAR User Manual Only now are the instrument specific commands loaded into the method editor They can be accessed by clicking on the ML_STAR toolbar in the toolbox window You can easily write the method by dragging icons from the toolbox on the left and dropping them in the method window on the right The resulting method will look like this Initialize on ML_STAR Channel 1 5 11111111 Optimized channel use All sequence position Sequence ML_STAR Vvastel6 Sequence counting 0 Manually Always initialize on Return value count 3 Load deck Load Carrier on ML_STAR Rack type pitcar lSmd_0001 Barcode file barcode_1 txt Return value count 4 Load Carrier on ML_STAR Rack type pitcar lSmd_ 0002 Barcode file barcode_1 txt Return value count 4 Load arrier on ML_STAR _ Rack type tip_car_ 460 0001 Barcode fi
9. Firmware is the set of instructions that are downloaded to the Microlab STAR by the manufacturer to enable the instrument to carry out its functions Normally the User Software is used to control the instrument However the Microlab STAR can be controlled directly with the CoCo firmware protocol CoCo means Communication and Control This is done by controlling actions of the Microlab STAR with single firmware commands Firmware commands are listed in the Service Manual for the use of Hamilton engineers servicing the instrument 3 5 Installation and Set Up The Microlab STAR will be unpacked and installed and initial set up performed by a trained HAMILTON technician The customer need only ensure that a suitable control PC is available for installation of the ML STAR User Software Make sure that the Microlab STAR is connected to a 115 to 230 V AC 50 or 60 Hz socket The Microlab STAR automatically recognizes any voltage within that range without user intervention Select the appropriate voltage at the pump station of the needle washer The needle washer does not recognize the voltage automatically Page 36 of 225 610766 02 Microlab STAR User Manual 3 6 Power Voltage 3 6 1 Basic Microlab STAR We recommend using a Uninterruptible Power Supply UPS for the Microlab STAR Ensure that the instrument is correctly earthed when connected to the power supply The mains plug is on the left hand side of the instrument towards th
10. MiStarlsWasteRack 0 O or 1 Into this rack tips or needles can be ejected TRUE OSFALSE MiStarTipRack 0 0 8 0 Standard Vol Tip disposable 1 Standard Vol Tip disposable with filter 2 Low Vol Tip disposable 3 Low Vol Tip disposable with filter 4 High Vol Tip disposable 5 High Vol Tip disposable with filter 6 Low Vol Steel Needle 7 Standard Vol Steel Needle 8 High Vol Steel Needle Page 200 of 225 610766 02 Microlab STAR User Manual NOTE At edit time the user does not have to define the tip type in the pick up tip or needle The information is taken from this property At edit time the user does not have to define the tip type in the eject tip or needle At edit time the user can only select sequences out of the list of sequences with underlying racks having these properties At edit time in the aspiration or dispense step the user can only select sequences out of the list of Sequences having underlying racks without these properties Needles can only be ejected into a rack with the property MIlStarlsWasteRack and the corresponding MLStarTipRack setting Tips can only be ejected into racks with the setting MiStarlsWasteRack e g standard waste Properties for wash station MiStarNeedleWashRack 6 8 6 Low Vol Steel Needle 7 Standard Vol Steel Needle 8 High Vol Steel Needle MiStarWashStationNumber _ tan Number of the wash rack 4 Wash Station 1 Rack 1 3 Wash Station 2 Ra
11. The needle wash station needs to be installed by a trained Hamilton engineer NOTE Do not use disposable tips with the wash station To use two different needle types two wash stations of different type are needed For a 16 channel instrument two independent wash stations are necessary to enable uninterrupted pipetting with one needle type A maximum of 2 wash stations each with 3 wash modules and its own pump station can be installed on one Microlab STAR The carry over of the wash station depends on the wash program Typical values are 10 to 10 The wash parameters can be set within the User Software prior to each wash step 610766 02 Page 29 of 225 Microlab STAR User Manual For each of the two wash solutions or wash and rinse solution a set of parameters can be specified The rinse time is the length of time liquid flows through the wash chamber The soak time is the length of time wash liquid is held within the wash chamber The flow rate is given in millilitres per second for both wash liquids For the drying of needles an additional parameter is used The draining time gives the length of time allotted for draining the liquid from the needles A set of default parameters is given for the wash process within the relevant dialog boxes of the User Software To empty the waste liquid container or to refill the wash solutions lift the container by the handle The valve at the bottom au
12. If Stop flow rate is equal to zero the plunger movement becomes gradually slower during the dispense until it stops e Stop back volume volume which is aspirated again immediately after the dispense as part of the dispense step This volume is aspirated automatically as quickly as possible The next tab defines the correction curve Edit Liquid Class Standard olume_Water_DispenseJet 1 0 Liquid Water Tip type Standard volume tip without filter Dispense made Dispense jet Correction curve Corrected in ul Poe Masimum liquid volume for New Entry specified tip type 0 0 0 20 0 ALU 100 0 200 0 300 0 350 0 Abbrechen Hite 610766 02 Page 103 of 225 Microlab STAR User Manual A correction curve has a target volume and a corrected volume The target volume is the volume to be dispensed The corrected volume is the volume plunger travelling distance x diameter that actually needs to be aspirated for this purpose In aspirate or dispense steps the target volume which will actually be dispensed into the vessel must be entered The corrected volumes are usually determined gravimetrically Accordingly a corrected volume of 105 ul does not mean that 105 ul of liquid will be dispensed When the tip is emptied 100 ul are dispensed The correction is mainly due to the properties of the air column above the liquid The definition of liquid classes allows you to pipette any given liquid
13. ML_STAR Tins 0 O U Oa ee 2 ustarsames ol of of ajm saree ol PF of of PD _ 610766 02 Page 137 of 225 Microlab STAR User Manual Click on add all sequences to load all sequences and corresponding carriers Select the Samples sequence to be reducible by clicking on show details Click OK Now add the pipetting step by dragging it to the next line MH Pipette with Tips Edit General Tip sequence Instrument short name ML STAR S _ Ren oad gt ML_STAR Tips Standard pipette procedure Aliquot procedure IY Reducible by user Pipette mode Tip handling replacement i Simple 1 1 C Replica 1 n i Pooling n 1 Multiplicity n After each dispense One tip set E Siter tranetered volume C None After each sample Volumes Pipette volume pl Tip counter a0 Pre aliquot volume pl Rest volume pl fo T Tip counter Write tip counter Redizpense height from bottom mm Disp to waste fa Disp to aspirate sequence Channel settings Error settings f Aspirate sequence is controlling Aspirate sequence MLS TAR Samples T Reload Reducible by user f Not reload F Aspirate all LLC Advanced Liquid liquid class Dispense sequence is controlling Dispense sequence MLS TAR Plate f Reload W Reducible by user Multiple use Dispense mode LLO Advanced tf Jet Surface
14. ML_STAR TransportSequencePlate10To13 Sequence counting 1 Automatic Return value count 3 ML_STAR TransportsequencePlatel Tos Reset Sequence after loop Then we loop over the transport sequence TransportSequencePlate1 To spanning all plates on the left hand plate carrier At the beginning of the method the plates are located on this plate carrier In step 3 we pick up the plate from the current sequence position of sequence TransportSequencePlate1 To Get Plate Edit Movement one Complex movement parameters UW to carne Retract distance Transport mode z 0 Plate only Lift up height Plate sequence ML_s TAR TransportS equencePlate Labware orentatiors Sequence counting 1 Automatic Lid sequence Advanced Error settings coed Heb The Movement Type gives 2 options either to pick up the plate from a carrier which is a simple motion or to pick up a plate from a reader etc which is a complex motion For a complex motion additional inputs can be made such as the retract distance and the lift up height see online help Transport mode specifies whether the plate is to be picked up with or without a lid or just the lid itself 610766 02 Page 167 of 225 Microlab STAR User Manual Sequence counting again determines whether the iSWAP returns to the same or to the next plate of the given sequence next time Note that a plate lid by
15. Microlab STAR User Manual 15 4 Walk Away Predefined Error Handling The user may define a walk away error handling which uses predefined default settings for different error situations These settings can be customized for single steps only For SMART steps the default error recovery is fixed For every instrument specific single step of your method an individual error recovery can be defined You can configure the appearance of the error recovery dialogs which buttons are available the default procedure which error is flagged in the trace file a timeout after which the default recovery is carried out the dialog automatically closes down For this purpose every instrument specific single step has a Error Settings button For the aspiration step it looks like this Aspirate Default Error Handling Pressure LLD Error Ho Tip Error Execution Error Hardware Error Not Executed Error Syrntas Error Improper Aspiration Error Clot Error Liquid Level Error M Use det aul Error description Liguid level not found or not enough liguid Exits defaut error handling user defined error handling is used if specified in method Timeout fo 7 z Infinite Sound fF M Abbrechen Hilfe Among these various tabs we have selected Liquid Level Error The default settings are activated 610766 02 Page 179 of 225 Microlab STAR User Manual To customize the settings disable t
16. Microlab STAR e g carriers racks tips steel needles and waste containers Commercially available liquid containers such as microtiter plates and tubes may of course be used If working with contaminated samples the user need not touch them The Microlab STAR will drop its used tips into a waste container that should be emptied as soon as it is full The Microlab STAR products conform to European norms as regards interference immunity However if the Microlab STAR is subjected to electromagnetic RF fields or if static electricity is discharged directly onto the Microlab STAR its Liquid Level Detection ability see below may be negatively affected It is therefore recommended that the Microlab STAR be kept away from other equipment that emits electromagnetic RF fields in the laboratory and that static electricity be minimized in its immediate environment 1 5 2 Electrical Safety Precautions Before removing a mechanical or electrical component the Microlab STAR must first be switched off and disconnected from the main electricity supply and PC 1 5 3 Biohazard Precautions If the Microlab STAR becomes contaminated with biohazardous material it should be cleaned in accordance with the maintenance procedures given in the section Maintenance 3 7 Observe and carry out the maintenance procedures given Failure to do so may impair the reliability and correct functioning of the Microlab STAR If working with biohazardous samples
17. Mis position 2 a mm Volume fo ul cov He In the upper part accept the default for using the same liquid class as in the aspiration step Click OK to accept the values Click OK in the dispense window NOTE Using a different liquid class for aspiration and dispense is allowable but not recommended Finally drag TipEject A dialog box appears Tip Eject New Sequence settings Use default waste 1 On Eject destination Sequence counting ML_5 TAR waste T UO Manually Channel settings Error settings OF Cancel Help Accept the defaults and click OK Page 132 of 225 610766 02 Microlab STAR User Manual NOTE Always use manual sequence counting for ejecting tips into the waste container The waste position is itself a sequence having just 8 or 16 positions Automatic incrementation would result in an error the next time tips are ejected into the waste no positions left over Finally the carriers have to be unloaded This is done by the unload command with the carrier name as a parameter Step UnloadCarmer Edit Carrier Carrier Click OK Repeat the unloading for all three carriers Within this method some comment lines have been inserted The dialog is simple Comment Edit Comment xl Cancel Help To enter a new line hold CTRL and press Enter Now your method is complete Exit the method editor by selecting File Exit from the
18. One tip set After fanstered volume None After each sample Volumes Volume per well pl Tip counter 2U Pre aliquat volume pl Fost aliguot volume pl 20 z0 Tip counter T Write tip counter Redispense height from bottom mm Disp to waste 90 Disp to aspirate sequence Channel settings Error settings Dispense sequence is controlling Dispense sequence MLS TAR Plate Me Reload Reducible by user f Not reload Dispense mode LLD Advanced f Jet i Surface i Asoiate sequence ie controlling Aspirate sequence MLS TAR Samples T f Reload W Reducible by user Multiple use F Aspirate all LLD Advanced Liquid liquid class water for aliquot Standard olume Water Aliquot et Cancel Help The selections made here are Aliquoting procedure The pre and post aliquots are 20 ul as well as the volume of the main aliquots The post aliquot is dispensed back to the aspiration sequence The tip sequence is ML_STAR Tips The tip handling is change after each sample meaning that all aliquots even if multiple repetitions are necessary are performed with the same tips The aspiration sequence is ML_STAR Samples and reloadable this has no influence in this example because the dispense sequence is controlling and one aspiration is sufficient to aliquot the whole plate The controlling dispense sequence
19. Page 164 of 225 610766 02 Microlab STAR User Manual Finally the AT Barcode Filter Program of the Microlab STAR user software is started by a library function from the HSLML_STAR lib library This function generates the Microlab AT like barcode file for the specified plate within the c barcodes directory P4 CreateATBarcodefile of HSLML_STARLib Edit Library name C Programme HAMILT ONSLibrarySHSLM L_STAALIB hal Warable Function name ri HSLM L_STAA Creates TB arcodetile F Function parameters Parameter Description ee ms SSSR z Labware id Plate 13 9 Using the iSWAP robotic plate handler The following example demonstrates the use of the ISWAP with ML STAR A set of plates is pipetted transfered to a barcode scanner position without opening the robotic hand and finally transferred to another carrier Note that the iSWAP works on the basis of sequences Plates are moved from one sequence to another The sequences remain fixed on the deck but the plates change sequences For this to happen target and source plate position must be of the same labware type For this example let s create the following deck layout using PLT CAR L5FLEX for the microplates Use the Add Labware dialog right click into the deck where no labware is located to add a plate position directly to the deck in front of an external barcode reader Enter the coordinates x 100mm y 350mm z 220mm Click OK 610
20. Suitable protective clothing safety glasses and protective gloves should be worn During Microlab STAR operation do not place hands in the way of moving parts or on the working deck Keep your head and hands away from the work surface of the Microlab STAR when it is in operation the pipetting arm and channels move fast and it is possible to sustain an injury In general never lean over the Microlab STAR when working with it When working with samples do not switch tubes around after they have been identified by the barcode reader This could result in incorrect test data When working with samples which will be used in particularly sensitive tests take into account evaporation and condensation that may occur while the method is running Perform test runs i with deionized water and ii with the final liquids prior to routine use Test for all the liquid classes you are going to use Liquid level detection needs to be explicitly tested when working with foaming liquids If sampling aggressive liquids use filter tips During operation the Microlab STAR should be shielded from direct sunlight and intense artificial light NOTE Discard used tips Do not reuse them Use filter tips for tasks which are sensitive to cross contamination aerosols Note that foam may affect the accuracy of liquid level detection As a general safety principle the inner diameter of sample tubes reagent vessels etc must be greater than the chan
21. observe and carry out the maintenance procedures paying particular regard to cleaning and decontamination Wear gloves when handling the pipetting arm and channels the carriers racks and containers and the tips and steel needles Avoid touching the discarded tips in the waste container Any surfaces on which liquid is spilled must be decontaminated 1 5 4 Computer Precautions Guard against software viruses Use only manufacturer s original installation CD ROM sets for the operating system and the original HAMILTON software Only the HAMILTON User Software and the firmware protocol CoCo KuSt cf Microlab STAR Service Manual may be used to control the Microlab STAR 610766 02 Page 9 of 225 Microlab STAR User Manual 1 6 Warranty Statement HAMILTON Bonaduz AG CH 7402 Bonaduz Switzerland is the manufacturer of the Microlab STAR The Microlab STAR is sold in accordance with the general conditions of sale of HAMILTON Bonaduz AG HAMILTON warrants this product to be free of defects in material and workmanship for a period of 12 months from the date of delivery ex works Bonaduz HAMILTON or the authorised HAMILTON representative will repair or replace at its option and free of charge any product that under proper and normal use proves to be defective during the warranty period HAMILTON shall in no event be liable or responsible for any incidental or consequential damage either direct or contingent HAMILTON accessories
22. process the pressure within the pipetting channel is measured in real time Analyzing the shape of the p t curve the system can distinguish the following situations A correct aspiration takes place Air is aspirated into the tip because for example the cup has not been filled properly A clot blocks the tip The aspiration monitoring can be switched on and off for each individual aspiration step of a method using the appropriate commands see section 2 4 For pressure based clot detection a threshold can be given in arbritrary A D analog digital values typically 100 A D Page 20 of 225 610766 02 Microlab STAR User Manual values The range of A D values of the pressure sensor is from around 800 at ambient pressure to lt 10 A D values for 18 mbar below ambient pressure For comparison the hydrostatic pressure of 100 ul of water in our standard tip is around 2 mbar p t ASP OK noo SS a p Thresheld Standard Tip 00 uf wwithou Fite Aspiration speed 100 uls ur ater 00 Volume 50 ul Tip blocked on C 100 200 300 400 500 B03 700 S00 300 1000 ASP Clot time ms Q Aspiration monitoring based on pressure NOTE Pressure based monitoring works only with unused disposable tips not with needles The volume range for the monitoring depends on the specific assay The lower limit is in many cases an aspiration volume of 50ul Pressure based monitoring has been optimized for aq
23. rack type PLT CAR_L5AC ml_star LoadCarrier Position 12 rack type PLT CAR _L5MD ml_star LoadCarrier Position 18 rack type TIP_CAR_480 Now we aspirate from one plate and dispense into another using new tips Under language double click or drag while to a line below the last load carrier command Page 206 of 225 610766 02 Microlab STAR User Manual In the method frame appears while Position the cursor between the parentheses and double click sequence under variable in the command frame In the dialog box select ml_star Source on the left and GetCurrentPosition on the right and click OK This results in while ml_star Source GetCurrentPosition Now add by hand the condition gt 0 while ml_star Source GetCurrentPosition gt 0 This means as long as the position of the source well is greater than zero or the operation is not finished do the following When the position reads zero the sequence has been worked through completely Now we enter the pipetting commands between the curly brackets Under the submenu ml_star double click or drag the single steps Now double click or drag tip pick up The same dialog box as in the graphical method editor appears NOTE Single steps are the same in the HSL and the graphical level of programming Choose the same settings as in the example with the graphical method editor The same is true for
24. 1 gt Sequence Position No 1 channel No 2 gt Sequence Position No 2 etc unless instructed to do otherwise A sequence such as the one shown below has three pointers or important positions which may change during a run The current position which is the first unused well in the current situation the end position the last position to be used for pipetting and the total number of elements the overall length of the sequence Consider this example MyPlate 1 2 3 4 5 6 7 8 9 10 11 12 T T j Current Pos End Pos Total Elements Within the method all three positions may be altered or requested by the appropriate functions A standard sequence is created automatically when the rack is placed on the deck Custom sequences can be defined graphically by selecting the appropriate wells in the Sequence Editor or on the fly within the programming of the method In methods defining the pipetting steps to be performed only the sequences and not the actual labware items are referred to Pipetting always occurs according to a sequence 7 1 Sequence Editing To edit or change sequences start the sequence editor by selecting Tools gt Sequence Editor in the Deck Layout Editor The Sequence Editor is illustrated on the next page When the sequence editor is started from the Deck Layout Editor a new untitled sequence is created automatically Add labware positions to this sequence by selecting clicking on positions wells in
25. 4 3 Structure ofthe User SOmw are sessssacsicdetceccciniactsscecseseviecasvisecbiaevoun astiseusiwrsearasseisdubesseersaneive 44 AA PNCCESS TRIG TS sai ast yack sadeedi casi ecgsccders a a O a E aa Ee 45 AD File SUPUCCURE senao RE A AE EE EA NE E EEE RRi 46 9 Tnstrument CONPIZUT ATION cessere ea eE EEES aA 47 6 Def mimo he Deck LAV OW sirina eana NE ANE E EER 50 Gol New Deck Layouten a a a O aa aa a Eia 5I Oll Save Deck Bayou rsissnnenn erore a a a a N R 52 6 1 2 Open Exis ns Deck Tay Outs inana eraa aa a E a aa AAi 52 6 2 Adding Labware to the Deck Layout eooosssssecccocsssssecccocsssseececoosssssecccoossssseceesossssesecesssssse 52 6 2 1 Adding Labware to Track Positions on the Deck nnsnssssessssseoooeeeeeessssssssssssssetererrssssssssssssseeeeeees 52 6 2 2 Adding Labware Directly tothe DECK eerren aint a 55 623 Seach aD wares an a E E pained 56 6 3 Making Changes to the Deck Layout cccccsssssssssccccccccssscssccccccccccssssssccccssceesssscsccooees 58 TEE a AT ILAE ATE EATA E ATE AA A A 59 Tok SSCGMCNCE Editino ssoosoeenoi n E EEE 59 Li Addie POSIMONS O d SEQUCHCO m a O A 60 KE Pli foaonea a saha those ton cesta as arlene tiie hand alana iat nae 61 DUN DON I ACK sar cee ca pecenctane gece taitesateeen tenes E teehee saeiaas ee adaiaetametn et cnces 61 TAA SOnting Dy GHG COMMING insenecserssosarseckonnsiedcaueubamancssehcodestodsasaniatoedmdaceagetsckeedautadsanehoataadentamanhscoese 61 Page 4 o
26. 610766 02 Page 175 of 225 Microlab STAR User Manual 15 3 Runtime Error Handling Prior to runtime error handling being used several types of problems causing errors have to be solved first Among these are Syntax errors when programming in HSL forgotten Logical errors tip eject before pick up asp 10ul disp 200ul Semantic errors wrong pipetting pattern Method deck interaction errors dispense 100 ul into the first well of a 1536 well MTP Liquid handling application errors droplets foam unpipetted wells User related errors sample tubes not filled completely wrong deck loading barcodes unreadable These problems cannot be handled by any runtime error handling Problems that can be handled in runtime are Not enough liquid liquid level not found if it occurs only rarely No tip picked up Clot detected Barcode unreadable if it occurs only rarely Execution error channel no 1 has an error e g not enough liquid then channel nos 2 8 have an execution error because they have been stopped before completion of the step NOTE In principle each channel may have one or more different types of errors at a time If all channels have the same error at the same time a colletive recovery can be made We now focus on some important examples A detailed description is available in the online help Click on Error Settings within the single step dialogs of the Microlab STAR specific commands Page 176 of 225 6
27. Computer Requirements The Microlab STAR is controlled by a dedicated user software program which controls all functions for daily work routine method programming running methods and other services The Microlab STAR and the computer controlling it may be linked in two different ways By a serial interface RS232 or By a Unified Serial Bus interface USB The Microlab STAR automatically recognizes the type of communication in use The Microlab STAR requires a recent model of PC including a mouse CD ROM drive high resolution monitor and graphics SVGA or better 1 2 serial ports for a USB about 1 GB HDD and 128 MB RAM and Windows 2000 service pack 1 or better as the operating system To avoid any loss of data we recommend a UPS uninterruptable power supply for the PC NOTE Microlab STAR s functioning has been verified using Windows 2000 exclusively Running the Microlab STAR under any other operating system than Windows 2000 may lead to severe problems and malfunction 3 4 1 User Software The Microlab STAR User Software is the controlling software for the Microlab STAR It is a Windows based menu driven interface allowing the user to define deck layouts and methods and then to run the Microlab STAR The Microlab STAR User Software allows the user to program and run different methods for aspirating and dispensing liquids NOTE Each programmed method has to be validated by the user 3 4 2 Firmware
28. Control OnePlateT oPlatePipette lay IOJ x File View Window Help cap fee OnePlateT oPlatePipette_lay tan ie ys 12 52 42 gt a1 aTEM Parse method start Method file C ProgrammeS HAMILTON Methods D emo 12 53 13 gt SYSTEM Parse method complete Single Step Pause Abort 0 processed approximately For Help press F1 Run Time Control window The relevant deck layout appears in the frame in the upper half of the window The lower half contains the Start button and another blank frame which will display the run method log method trace file as it generates 610766 02 Page 171 of 225 Microlab STAR User Manual Press the Start button to run the method You will see that the steps in the method are traced to the log frame A loading dialog appears requesting a reduction of the number of positions on the source plate as well as a start position for the tips to be picked up Both pieces of information are optional Load Sequence Miel x Load the the following carriers olt_ car l6rod_O001 plt_carlSmd_O002 tip_car_480_ 0001 Sequence ma Lest Remove all Removed Remaining sore a o Je o y O a S o E a e ao me o o Od O O o O o o Reset Remove All je je Help You may enter the number of wells on the source plate for this run delete wells graphically from the sequence or accept the default copy the whole plate Manual Load Only Load the deck with the carr
29. Page 217 of 225 Microlab STAR User Manual SMART steps 5 6 37 79 80 81 82 93 94 95 106 123 131 133 148 155 158 169 173 200 Software 1 4 8 11 13 21 29 30 36 38 41 42 46 49 50 51 52 54 59 64 77 82 107 111 112 119 127 131 155 174 184 203 sorting 5 68 69 70 71 72 74 Specifications technical 4 47 Teaching box 55 Teaching mode 4 55 64 65 Tip counter 6 53 174 Tip eject 3 20 95 106 141 197 Tip pick up 81 84 94 106 141 143 144 Tip rack 15 29 59 60 61 81 93 132 143 149 151 155 159 160 174 207 218 Tips disposable 3 6 9 10 11 14 15 16 17 18 19 20 21 22 23 25 26 27 29 30 33 35 47 48 53 55 59 60 61 62 65 80 81 82 83 84 88 91 93 94 95 97 106 114 115 116 131 132 135 140 141 143 144 145 147 149 151 153 155 157 159 160 Page 218 of 225 172 174 178 179 180 181 183 192 197 206 207 209 218 Tolerance 187 Track 4 55 59 60 62 64 120 149 216 217 Training 4 46 Tubes 6 9 10 12 15 16 18 28 29 30 31 32 33 55 59 60 62 80 82 83 85 86 87 91 148 149 151 154 155 169 174 197 198 207 209 216 Variables 5 15 18 76 78 82 100 101 103 104 107 108 124 126 140 163 165 166 167 178 179 180 183 Verification 4 46 47 Walk away 90 200 202 Wash station 15 20 32 34 35 36 42 45 81 93 94 97 98 169 Waste s
30. and the clearance height here 80 mm at which the pipetting arm can pass over the container without touching it as measured from the container base The maximum pipetting height counted from the container bottom is 4 mm because we want to allow the tip to go down to a position of 4 mm above the tube bottom this gives the dead volume The touch off height means the position of the tip when dispensing with touch off into an empty container It is set to 0 3 mm although this option is not available for all instruments including the ML STAR at the present time The same is true for the wick side of container checkbox which in principle enables or disables touch off at the sides of the containers but is also not available for all instruments The next important option is the Liquid Level Valid checkbox which is activated here and the value of 75 mm above the tube bottom to start the liquid level detection within the tube The Thickness of Container base is used to place a rack filled with such containers in the correct Z position Page 186 of 225 610766 02 Microlab STAR User Manual When the values are selected click Next to open the next dialog box for segment definition Segment Definition from container rim to base Rounded base segment VeAlbone bare segment Select cylinder as a shape for the upper segment and fill in the values for the inner diameter 10 mm and the segment heig
31. be helpful sometimes This can be done by modifying the configuration To integrate the vacuum box in the configuration of the ML STAR you have to do the following open the Hamilton Deck Layout Editor window and select Open from the File menu Then open the Config folder and change the Files of type to Deck Template Look ir Sy Contig e Fa aa ML_STAR tpl ML_STAR2 tpl YacBox tpl File name Open Files of type Deck Template tpl Cancel A From the three different available deck layouts select the ML_STAR2 tpl and open it Select the vacuum box under Add labware and put it on the right place on the deck Select File and click Save Your default Microlab STAR deck layout is now changed Whenever you open a new STAR deck layout the vacuum box will already be placed on the deck Close the Hamilton Deck Layout Editor and open the Hamilton Method Editor Select New under the File menu and save your method with a new name Click on Instruments under the Method menu Page 108 of 225 610766 02 Microlab STAR User Manual Click on the grey Deck Layout File button associated with your Microlab STAR instrument A list of deck layouts will appear Select the Microlab STAR deck layout you wish to associate with this method The Layout window reappears with the selected layout file showing Afterwards select the VacuumBox instrument
32. channel use on Waste destination ML_STAR Waste Sequence counting 0 Manually Return value court 1 End Loop nunc_96 flat_l Reset Sequence after loop A simple method Inserting a new command or double clicking on an existing command opens the appropriate dialog to edit the command specific parameters for example the Assignment command used to define and assign variables BA Assignment Edit Varable Value volume 150 Cancel Help Assignment parameter dialog 8 4 Editing a Method A method can be edited by the usual Windows copy and paste techniques Mark the step or the block of adjacent steps within the method editor and right click the method window to access the cut copy paste menu To insert a step from the buffer click on the line above which the step should be inserted and then paste 610766 02 Page 89 of 225 Microlab STAR User Manual 8 5 Variables Within a method the user may typically wish to define and process variables The Graphical Method Editor recognizes three types of variables integers numbers floats floating point numbers and strings text variables Variables can be newly defined within the dialogs if they occur e onthe left hand side of an equation e g x 0 or x t 1 or e within edit fields that hold only one variable If not explicitly requested as is the case in the Open file dialog the variable types are identified automa
33. fa tt a a 1 4 2 Dimethyl sulfoxide Pa a fatal o f a ala Ethyl acetate Lae e dakal a a a we Herne ot a ft fo ile a a a Hydrochloric acid 20 Pa a4 fa tat a ilaia i Isopropyl alcohol Pa a halal a hal a dala Methanol afa Po tat ff ailai Methylene chloride Pa a alate fs 1 i a Nitricacid 510 1 2 14 ia 1 f a i ae Nitric acid 70 Pa 4 felts a al 1 4 a Phosphate buffer Pa a4 fata o 4 1 4 a Phosphoric acid 85 fe as ata o af a a Potassium hydroxide conc Pa a fa tat a a 1 2 8 Sodium acetate 1 t 3 14 oo tf a i a Sodium borate Pa a4 fata olila Sulfuric acid 1 75 Pe akalala lalala lel Urine afa Pa fa ta a ailai Triethylamine Pa 4 lalalo lal 1 3 a Toluene Late 4 bed 4 lalake kal Page 224 of 225 610766 02 Microlab STAR User Manual Effects Key to codes in above table no effect little or no noticeable change moderate corrosion or other change in physical properties or dimensions not recommended for continuous contact severe corrosion or physical change prolonged contact not recommended 2 slight corrosion or discoloration No data Materials 1 4310 Steel EPDM Ethylene propylene elastomer FPM Fluoroelastomer PE Polyethylene Storage Containers for Wash Liquids 610766 02 PEEK PP PTFE PVDF SI Polyetheretherketone Polypropylene Polytetrafluorethylene Polyvinylidenefluoride Silicon Page 225
34. from the current Z position Click OK to accept the values Click OK in the aspirate window Now drag Dispense to the position below the aspirate command A dialog box appears Step Dispense Edit Sequence Sequence counting 1 Autornatic Volume Dispense mode 50 ul Individual volumes i Jet mode empty tip Liguid level detection LLO Capacitive LLD sensitivity Submerge depth Fis height from bottom 0 Off 2 2 Channel settings Advanced Error settings Cancel Help Choose ml_star Target as sequence to dispense to the target plate Choose sequence counting automatic Enter 50 ul for the volume again Select the dispense mode Jet Mode Empty Tip Select off as the capacitive LLD setting because the target plate is empty and enter 2 mm as liquid level corresponding to a height of 2mm for the dispense measured from the container bottom 610766 02 Page 131 of 225 Microlab STAR User Manual Click Advanced A dialog window appears The lower part of the dialog is exactly the same as for the aspiration step liquid following and mixing settings Dispense Advanced Same liquid class as first aspirate of cycle Filter Tip type Dispense mode Standard volume tip without filter Dispense jet Liguid class Liquid following during dispense and mis 1 On Preninsing Mix settings Cycles 0 Off
35. head from the instrument dependent modules The general modules with a common look and feel for all HAMILTON instruments include e Deck layout editor for mapping the position of labware on the deck e Method editor for programming methods e Sequence editor for defining pipetting sequences e Labware editor for defining custom labware to be used in deck layouts e HSL Method editor for programming methods using text entry mode e Run execution for running methods on the instrument The general tools include e Version info The Microlab STAR specific editors include e Liquid Editor for defining liquid classes e Configuration Editor for configuring the User Software to a particular instrument The Graphical Method Editor lets the user write methods in a graphical syntax free environment Technically this editor is a code generator producing a low level code which is then interpreted by the run control This low level code is called HSL Hamilton Standard Language because it provides a common programming approach for all Hamilton instruments HSL can be accessed directly using the HSL Method Editor HSL is a syntax dependent language that offers even more flexibility than the Graphical Method Editor The instrument specific steps e g aspiration dispense see below are the same for the Page 44 of 225 610766 02 Microlab STAR User Manual graphical and the HSL method editor The general syntax used in the HSL
36. in order to get all the specific vacuum box commands Instruments Smart Steps Mw IML STAR Smart Steps OK Cancel Help 610766 02 Page 109 of 225 Microlab STAR User Manual After clicking OK three new sets of commands are now accessible via command bars in the lower left of the Hamilton Method Editor window namely the Microlab STAR specific commands ML_STAR and ML_STAR Smart Steps and the vacuum box specific command VacuumBox O x File Edit lew Method Tools Window Help 0g A O dt I az B Y acuumBox med y E Ee General ML STAR Smart Steps Ready a Page 110 of 225 610766 02 Microlab STAR User Manual 12 2 Commands for the AVS There are three commands associated with running the vacuum box These are in alphabetical order and are described below Vacuumbaox Empty waste ply Vacuum boxe number fi Duration E0 seconds OF Cancel Help This command turns on the waste pump inside the vacuum box controller so that the waste tray in the conditioning chamber can be evacuated Select the vacuum box to evacuate Vacuum box number 1 is the default The pump duration will need to be determined by trial and error Generally 25 seconds are adequate for emptying a full waste tray 200 ml If the waste bottle is full when this command is running an error dialog box will appear The user then can empty the waste bottle and continue the
37. is the C C notation and associated language constructs 4 4 Access Rights The User Software does not define any user access resirictions However specific access rights may be restricted to trained users within the operating system or by applying write protection to specific files 610766 02 Page 45 of 225 Microlab STAR User Manual 4 5 File Structure The installation generates the following default directory structure if not requested otherwise Folders sels 3 Floppy 4 aS C eed J capture H E Multimedia Files lg NOV H E NT i e Frogram Files 4 Common Files HAMILTON Ate Measure tre te Measure stp ate Measure med ee Measure lay Ate Measure hsl Volume tre Volume stp Volume med Volunne lay Volume hs Tips4heedles tre Tips4hleedles stp Tips Needles med Tips4h eedles lay Tips Needles hel teslIPS stp teslIPS med testlPS lay testwPS hs H a p A E Labware C ML STAR_1 0 uf SWAF Note in particular the file contents of the following directories Bin Executables DLLs Config Configuration Files Labware Labware Definitions Logfiles Com Traces Port Trace Tip Counter File TipCount xls Methods Methods med hsl Deck Layouts lay Method Traces trc Library HSL libraries of commonly used additional functions c barcodes In addition this directory is used by the sample tracker to store Microlab AT like barc
38. itself has a sequence with 2 positions Selecting the transport mode Plate with Lid will increment the plate as well as the lid sequence Under Advanced additional settings may be chosen Get Plate Advanced E SWAP settings Grip force 5 Grip force 5 Tolerance 4 5 mm Grip height 3 mm Inverse grip 0 OFF Cancel Help Grip force determines the force that is used to grip the plate The tolerance gives a bandwidth in mm for the closing mechanism of the robotic hand in which the plate must be gripped torque sensor of iSWAP responds The grip height is the distance that iSWAP moves down from the plate s upper rim to grip the plate Inverse grip instructs the robotic hand to pick up a plate with the opposite orientation of the iSWAP this is not always possible The plate is now moved to the position of the barcode scanner the position of the sequence PlateAtExternalBCScanner without opening the hand Move Plate Edit E Move to sequence ML STAR PlateAte sternal CS canner Error settings omo o The final step places the plate on the right hand plate carrier Place Plate Edit Movement type Complex movement parameters Retract distance Transport mode 0 Plate only Y Lift up height Plate sequence M L_5TAR Transports equenceFlate Labware orientation Sequence counting 1 Automatic Lid sequence Page 168
39. like barcode file the execution of a Gemini worklist the formatting of an ML STAR barcode file and the deletion of an ML STAR barcode file Refer to the online help for more information on these functions The libraries available include Library This library function handles HSLArrLio array variables HSLDevLib devices HSLDIgLib dialogs HSLErrLiob errors HSLFilLib files HSLMthLib mathematical functions HSLSegLib sequences HSLStrLib string variables HSLTimLib timer HSLTrcLib trace files Page 92 of 225 610766 02 Microlab STAR User Manual 8 7 Binding Instrument Step Return Values If an instrument step returns information it can be bound to a variable by opening the Bind return values dialog select step of interest and select menu Edit Bind return values For example the get temperature step of the temperature controlled carrier returns the temperature and other information Bind return values Bind return values to vanables coed e Bind Return Values dialog These pieces of information can be stored in a variable for further processing or output The dialog generally lists the return values each with an editable field to define the variable the return value is to be bound to 8 8 Working with Worklists 8 8 1 File Formats Frequently you will want to import and export worklists The open file command of the Method Editor is designed to enable such file handling The data source type form
40. menu Chapter 15 Running the Microlab STAR explains how to run a method 610766 02 Page 133 of 225 Microlab STAR User Manual 13 4 Create a Method to Copy from Tubes to Plates Using SMART Steps This method copies tubes i e aspirates liquid from tubes in a carrier and dispenses them to wells in a microtiter plate The maximum number of tubes to be processed is 96 corresponding to a maximum of four 24 tube carriers 1T First create an appropriate deck layout In this case it will be Tubes ToPlatePipette lay To create this deck layout start the Deck Layout Editor by double clicking on the appropriate icon Select New from the File menu Select Microlab STAR as the current instrument see method editor description Click OK Now you see the schematic view of the Microlab STAR deck Create a deck layout as shown below 35 40 45 Deck Layout TubesToPlate Double click on one of the tracks In the pop up window you may enter a name standing for the tube carrier and click Browse Select Car24_cup15x100 rck as rack type for a carrier holding 24 tubes of 15 mm diameter and a height of 100 mm from the ML_Star directory Click OK The carrier is added to the deck layout Repeat this procedure three times more for the other tube carriers Place them in adjacent positions on the deck Now you have added the tube carriers Double click on another track to add a standard plate carrier A d
41. not set no load is required the carriers are fixed on deck or already loaded If MlStarCarlsLoadable is set a load dialog is shown and if MlStarCarlsAutoLoad is set loading is executed by the instrument 17 4 3 Information for Special Units Properties for calibrate carrier single command Carrier 1536 MiStarCarCalibratex 0 1mm __ Distance between left margin of po farrier to middle of measuring hole Distance between carrier front to middle of measuring hole 610766 02 Page 199 of 225 Microlab STAR User Manual NOTE If a carrier can be calibrated all values should be set The origin ts the zero point of the carrier front left down If the carrier cannot be calibrated these keys and values should not be exist remove key At edit time in the calibrate step the user can only select carriers templates with underlying carriers having these properties Properties for Temperature Controlled Carrier TCC MiStarCarls Temperated 0 O or 1 The carrier can be temperature controlled 1 TRUE 0 FALSE stations 1 or 2 MiStarCarlncubatorNumber 1or2 Number of temperature incubation NOTE There are only two TCCs allowed on a deck Each TCC must have its unique identifying number At edit time in the incubator step the user can only select sequences out of the list of sequences with underlying racks having these properties Properties for tip and needle handling
42. of HSLTipCountingLib Edit Library name C Programme HAMILT ONSLibrarysHSLT ipCountingL ib hel VWarable Function name TipCount Edit P Function parameters Parameter Description SeQuence ch L STAR AITI pS q4 pCountervariakle devicelontext iment L STAR 4 J 4 The TipCountingEdit function opens up a dialog on runtime showing the tip sequence and input fields for start and end positions Setting the start position to 1 will reset the tip counter then tips have to be reloaded manually This function has parameters the tip sequence the name of the tip counter which is stored in the variable TipCounterVariable here the device context select the only choice ML_STAR from the dropdown field and a timeout This timeout is used to close the 610766 02 Page 161 of 225 Microlab STAR User Manual dialog automatically without user intervention after 7 seconds in this case by defaulting to the current position of the tip counter Just for demonstration and not part of this method another step could be to simply read the tip counter et Read of HSLTipCountingLib Edit Library name C Programme HAMILTONSLibrany HSLT ipCountingLib hel Variable Function name gt TipCount Read P Function parameters Parameter Description Sequence lb ML STAR AllTips tiplounter Tiplounter y ariable The parameters are the tip se
43. on the deck at the beginning of the loading process The TCC will be installed by a trained Hamilton service engineer A maximum of 2 TCCs can be placed on one Microlab STAR 3 3 5 The Automated Vacuum System AVS Page 32 of 225 610766 02 Microlab STAR User Manual The Automated Vacuum Box System consists of a vacuum box a vacuum controller and the controlling software The Vacuum Box is fitted to the deck of the Microlab STAR and is controlled by the workstation s computer For this integration a special deck has to be mounted on the Microlab STAR The vacuum box accommodates a wide variety of 96 well filter and collection plates and is automated so that the filter plate can be transported from a waste conditioning chamber to an elution chamber The vacuum box is compact and a maximum of two can be used in parallel on the Microlab STAR deck with the same controller Filter plates can be placed onto or removed from the system by hand or with the help of a robotic arm see the section on iSWAP below The vacuum controller allows for user defined vacuum settings The software integrates seamlessly with MICROLAB Vector Loading is handled with a step in the method We now turn to a description of the components of the automated vacuum box system Vacuum Box The picture on the left provides a bird s eye view of the vacuum box showing the conditioning chamber on the right and the elution chamber on the left which is co
44. on the left and dropping them in the method window on the right The resulting method will look like this S OnePlateT oPlatePipette med ML_STAR Source ML_STAR Target ML_STAR Tips qh Pipette with Tips from ML_STAR Smart Steps We keg Instrument short name ML_STAR Standard pipette procedure Mode Simple 1 15 Pipette volume HI 50 Aspirate sequence WML STAR Source controling sequencey Dispense sequence WL_STAR Target J Unload from ML_STA amp R Smart Steps Instrument short name ML STAR unload F sequencers MIL STAR Tips ML_STAR Target ML_STAR Source Note Explicit loading specifying loading commands within the method is recommended for safety reasons but is not mandatory If no loading commands are specified no check of the carrier positions is performed and the user has to ensure that all carriers are positioned manually on the correct tracks This holds true for manual load and autoload Microlab STARS 610766 02 Page 119 of 225 Microlab STAR User Manual First drag the SMART Step load into the method window Instrument short name ML_ STAR ial Sequence sh Show details a EE hirer wor o BL STAR Ti A or oo d Add all sequences Add Remove Carrier calibration Error settings Channel number W Calibrate if supported E OF Cancel Help Click on Add all sequences to make sure all carriers are going to be loade
45. operate the Microlab STAR correctly and safely To achieve that aim Chapter 3 of the manual will describe the different components of the Microlab STAR and how they work Then in succeeding chapters we will describe what can be done with each the basic operations of aspirating and dispensing liquids The manual describes both the hardware and software of the Microlab STAR to the extent that a user needs to know them in order to operate the instrument After introducing you to the various parts of the Microlab STAR we show you step by step how to perform typical operations using those components Sample methods for typical applications guide you through the programming When you have worked through this manual you should be quite well able to operate the Microlab STAR Warnings and notes are included in this manual to emphasize important and critical instructions They are printed in italics in the left margin of the page begin with the word Attention accompanied by the symbol or the word Note as appropriate This manual refers to User Software release 3 0 for the Microlab STAR 1 2 Additional Microlab STAR Manuals A detailed software reference for the Microlab STAR is to be found in the online help of the User Software This online help will answer any question you may have about details of the Microlab STAR User Software The manner in which the Microlab STAR and its components are to be serviced is described in t
46. plates or tips MD medium density 96 or 384 well microplates HD high density microplates 1536 AC 96 well archive plates Ann identifies the part number revision e g A00 Example PLT CAR L5MD AOO is a carrier for 5 medium density 96 or 384 well microplates in landscape orientation A carrier must always be identified e g in deck layouts and methods by the unique descriptor with which it is tagged 3 3 2 Disposables CO RE tips come in 3 sizes low volume standard and high volume approximately 10 300 and 1000 ul respectively One blister pack contains 5 racks of 96 tips giving a total of 480 tios CO RE tips are available in boxes of 4000 6000 depending on the tip size CO RE tips are produced under sterile conditions i e the tips are RNAse and DNAse free Reusable needles with 50 300 and 1000 ul volumes may be used instead of tips Only HAMILTON needles and disposable tips should be used for coupling to the pipetting channel of the Microlab STAR The needle wash station is a device for the washing and drying of steel needles parallel to the pipetting process The wash station has the width of a normal microplate carrier 6T and is mounted on the deck of the Microlab STAR All tubing and electrical connections are routed through the docking station at the rear of the instrument Along with each wash station comes a pump station with three reservoir containers for wash solutions and a waste container This pump sta
47. repeated to length of Seq Asp Use Copy 2 plates into 1 after another With reloading of Seq Disp Transfer available tubes to as many plates as needed Case 2 Seq Asp is reduced to length of Seq Disp Use Transfers from many tubes to a plate and stops when plate is filled With reloading plate sequences Automatically fill as many plates as needed with liquid from tubes Case 3 Seq Disp is reduced to length of Seq Asp Use Transfer a variable number lt 96 of tubes into a 96 well plate left partly filled Case 4 Seq Asp is repeated to length of Seq Disp Use Transfer reagent from a container or e g 8 tubes to a complete plate Copy tubes repeatedly into plate With reload of aspiration sequence Automatically reload tubes until plate is filled The 4 cases are illustrated in the following diagrams Page 74 of 225 610766 02 Microlab STAR User Manual Case 1 L Sasp gt L Sgisp Control ASP Source 1 Source 2 The target sequence is repeated to match the length of the source sequence Another example for this case All Tubes to Plate s Reloadable plate sequence to load all tubes to as many plates as needed Case 2 L Sasp gt L Sdisp Control DISP axalelalelalalelelalarlearerlelealalarlavtalalarlalale aralelalelervarlelelalarlerlarlealalalarlavlalarlarlalale axalerlalelarxerlelealalarlerlerlalalarlerlavlearlerlarlalale aralelalelarx
48. sample are done with the same tip with one tip set or without tip handling none In this case STAR will have to pick up tips prior to the pipette step and eject them after pipetting using single steps Only for replicas and pooling the option after transfered volume becomes active This allows you to use fresh tips even for multiple aspiration and dispense cycles being performed with the same sample Finally a tip counter can be specified which enables the user to start with a set of tips partly used in previous runs at the correct position To read a tip counter it can be specified within the SMART Step load see that section A tip counter is specified by a name e g MyFirstStandardTipCounter Note that the name has to be placed between quotation marks Within the SMART Step pipette the current position of the tip sequence will continuously be stored under the name of the tip counter if the check box write tip counter is checked Note that a tip counter has to be written if it is to be read by the SMART Step load within the next run The next step is to define the volume s for the pipetting process For the SMART Step pipette one volume for all channels has to be defined Volumes Pipette volume pl 100 Pre aliquot volume pl Rest volume pl o U Redispense height from bottom mm Disp to waste 0 Disp to aspirate sequence lf a volume for simple transfers
49. that this requirement is only for generators which operate at over 10 000 Hz In Vitro Diagnostics The Microlab STAR is not intended specifically to be used as an In Vitro Diagnostic Device The following text defines an In Vitro Diagnostic Device from Directive 98 79 EC of the European Parliament and of the Council of 1998 10 27 on in vitro diagnostic medical devices in vitro diagnostic medical device means any medical device which is a reagent product calibrator control material kit instrument apparatus equipment or system whether used alone or in combination intended by the manufacturer to be used in vitro for the examination of specimens including blood and tissue donations derived from the human body solely or principally for the purpose of providing information concerning a physiological or pathological state or concerning a congenital abnormality or to determine the safety and compatibility with potential recipients or to monitor therapeutic measures Specimen receptacles are considered to be in vitro diagnostic medical devices Specimen receptacles are those devices whether vacuum type or not specifically intended by their manufacturers for the primary containment and preservation of specimens derived from the human body for the purpose of in vitro diagnostic examination Products for general laboratory use are not in vitro diagnostic medical devices unless such products in view o
50. the deck layout view The standard left click shift left click control left click and rubber banding rectangle are available for selecting positions in the deck layout view As positions are added to the sequence they are displayed in the grid in the left pane of the window The grid contains one row for each labware position in the sequence The order of the rows in the grid can be changed by standard cut copy paste drag drop and delete operations thereby changing the order of the positions in the sequence The grid has a fixed width X Y and Z columns may not be initially visible Moving the vertical splitter or the horizontal scroll bar will allow these columns to be seen Sequence information cannot be typed directly into the grid 610766 02 Page 59 of 225 Microlab STAR User Manual Additionally the functions of the sort filter panes can be applied to the selected rows of the grid if selected or all grid contents Once the labware positions in the sequence are in the desired order save the sequence Saving fixes the order of each row in the grid as the order of the labware positions in the sequence On saving you are requested to give a name to the sequence Any existing sequence in the deck layout can be activated for viewing and or editing by selecting the sequence name in the drop down menu Additional labware positions can be inserted in the active sequence by selecting them in the deck layout view Existing positions ca
51. the first line of the parallel process Vacuum box number fi Move to Move the carriage of vacuum box number 1 to the conditioning site OK Cancel Help Click OK Now set the vacuum parameters Yacuum Parameters Vacuum bos number fi Chamber Vacuum Inches of Mercury Ramp duration fio EE Apply a vacuum of 5 inches of mercury on the conditioning site for a total of 40 Vacuum duration fzo seconds seconds 10 seconds ramp duration plus 30 seconds vacuum duration Error Recovery OK Cancel Help 610766 02 Page 115 of 225 Microlab STAR User Manual Now empty the waste of the vacuum box Empty Waste Parameters Vacuum bos number fi Duration 20 seconds Empty waste on the conditioning site for UE Cancel Help 20 seconds Finally end the first parallel process iz End Parallel Edit E Name of parallel process process X Timeout W Infinite End of parallel process1 with all the vacuum box commands Timeout 3 g OK Cancel Help The bifurcation within the method editor ends after this step To process a plate for example with the pipetting channels of the microlab STAR add pipetting steps to the normal process left side of the bifurcation Page 116 of 225 610766 02 Microlab STAR User Manual 13 Methods for Microlab STAR 13 1 Overview Most of the instrument s day to day operations will b
52. the following aspiration dispense and tip eject steps We should now see the following lines in the programming window ml_star TipPickUp Channel 1 8 o0000000 Sequence ml_star Tip Tip type StandardVolumeT ip This command picks up standard tips without filter on all used channels from the sequence of the tip rack i e from the tip rack ml_star Aspirate Channel 1 8 o0000000 Sequence ml_star Source Liquid name StandardVolume_Water_DispenseJet Volume 50 ul LLD settings Capacitive on Pressure off Liquid following on This command aspirates liquid on all used channels from the sequence of the source plate i e from the source plate The liquid name specified enables the software to use the appropriate parameters Volume and the settings for liquid level detection are further parameters Here we aspirate following at liquid level ml_star Dispense Channel 1 8 oo000000 Sequence ml_ star Target Liquid name StandardVolume_Water_DispenseJet Volume 50 ul LLD settings off Liquid following on This command dispenses the given volume into the sequence ml_ star Target i e into the target plate The liquid name has to be the same as for the aspiration Here we dispense into an empty plate therefore LLD is switched off and we dispense in a jet following from 2 mm above the plate bottom 610766 02 Page 207 of 225 Microlab STAR User Manual ml_ star TipEject Channel 1 8 oo00000
53. to be fully functional Ensure the correct barcode orientation for tubes and plates see picture on previous page 3 3 Accessories Accessories are defined as additional components that can be ordered later by the customer and installed either by HAMILTON personnel or by the customer They include carriers a wash station a temperature controlled carrier and an automatic vacuum system which are described in this section of the manual Note that the availability of accessories is subject to change Please ask for a current list Microlab STAR Wash Station Pump Station with 3 wash af solutions and waste container A L Heat Exchanger for TCC en at Controller for Vacuum Box The Microlab STAR with Carriers Wash Station Temperature controlled Carrier and Vacuum System 3 3 1 Carriers The labware is placed on special carriers which are loaded onto the Microlab STAR HAMILTON provides a wide range of standard carriers for microtiter plates tubes tips etc All standard carriers can be added to the deck by the user The naming of carriers follows a systematic nomenclature X Y Z Ann X stands for the type of labware placed on the carriers e g TIP tips PLT plates SMP samples Y CAR for carrier 610766 02 Page 27 of 225 Microlab STAR User Manual Z describes the labware details e g L landscape orientation P portrait orientation Number number of items placed on the carrier
54. to run or from tube carrier to tube carrier 610766 02 Page 73 of 225 Microlab STAR User Manual An important issue is to select the controlling sequence This is either the aspiration or the dispense sequence As long as both sequences are of the same length this selection does not influence the pipetting if no pipetting error occurs Sequence handling in Pipetting Mode Simple But think of the situation where the aspiration sequence comprises e g 8 tubes and the dispense sequence is the one of a 96 well plate What to do now with the 88 remaining positions within the plate If the aspiration sequence is controlling the dispense sequence is reduced to the length of the aspiration sequence and the pipetting stops after dispensing into the first 8 positions of the plate sequence If the dispense sequence is controlling the aspiration sequence will be repeated until it reaches the length of the dispense sequence This results in multiple 12 for an 8 channel instrument transfers from the same 8 tubes to fill the complete plate Then the controlling disoense sequence is finished and pipetting stops NOTE The SMART Step pipette always equalizes the length of aspiration and dispense sequences The following table gives an overview of the different situations and examples L is the length of the sequence L Sasp L Saisp L Sasp gt L Saisp Aspiration Controlled Dispense Controlled Case 1 Seq Disp is
55. until a wash module is ready picks up needles from a wash station or from a separate needle rack which is in fact an empty tip rack filled with needles e g the lowneedle_ rck Dragging the command to a method tree opens the following dialog Needle sequence Channel settings Error settings Cancel Help The sequence requested here is the sequence running over all three modules of a wash station thus having 24 positions 8 from each module This sequence has to be generated using the sequence editor The SMART Steps automatically take care of handling the individual wash modules by incrementing the appropriate sequences in the background NOTE For 4 and 12 channel instruments an appropriate sequence has to be created with the sequence editor Thus for a 4 channel instrument the sequence should hold positions A1 A2 A3 A4 B1 B2 B3 B4 C1 C2 C3 C4 where A B C are the different wash modules Page 82 of 225 610766 02 Microlab STAR User Manual The needle eject command places the needles into the next available wash module and starts the wash process on request fal Heedle Eject New Instrument short name Needle sequence M Start wash Channel settings Error settings Cancel Help For both commands channel patterns can be set manually The error settings are again similar to those of the SMART Step pipette An additional SMART Step is available to set the w
56. with high accuracy Liquid classes are also available from HAMILTON s Application Engineering Group for custom purposes if you need them NOTE The standard liquid classes supplied with the instrument have underscores _ in their file names These liquid classes cannot be changed by the user They can be copied saved under a different name and then edited 11 3 Defining a Custom Liquid Class To define a custom liquid class select a pre defined liquid in the Liquid Editor start window and click Create The Copy Liquid window pops up Copy Liquid Standard olume_ Water _DispenseJet 1 0 New liquid class name My ewLiquid cence _ Copy Liquid window Type in the name of the new custom liquid class and click OK Back in the Liquid Editor start window select your new liquid class and click Edit Now all the parameter input fields in the Liquid Details window are active so you can make whatever changes you require Page 104 of 225 610766 02 Microlab STAR User Manual 11 4 Importing and Exporting Liquids A liquid is stored in a configuration file cfg Opening the Liquid Editor loads liquids defined in the standard liquid configuration file Hamilton Config ML_STARLiquids ctg If other liquids are needed from another liquid configuration file they can be imported Click Import liquid s in the Liquid Editor start window to display the Import Liquid s window Import Liquid s E4 Import fror
57. you have added a 96 well plate from Nunc to the carrier Repeat this to place the target plate on the other plate carrier Type Target in the dialog box Now that this plate type has been specified within the deck layout you may select it directly from the Type dropdown field Double click on one of the sites on the tip carrier A dialog box appears Leave the name field blank and click Browse Select standardtip_ rck from the ML_ Star directory Click OK Note In the example on the CD a second tip rack is added Select Save from the File menu to store the deck layout under the name OnePlateToPlatePipette Page 118 of 225 610766 02 Microlab STAR User Manual Click on Edit Method to open up the Graphical Method Editor From the method editor s Method menu select Instruments to link the deck layout to the method as described in the chapter The graphical Method Editor A window pops up Instruments X Short instr Hame Deck Layout File Low Level sil E Microlab STAR iMiL_STAR OnePlateToPlatePipett fHidden Choose OnePlateToPlatePipette lay as the deck layout browse by clicking the button and click OK Only now are the instrument specific commands loaded into the method editor They can be accessed by clicking on the ML_STAR SMART Steps toolbar in the toolbox window You can easily write the method by dragging icons from the toolbox
58. 0 85 with no condensation 610766 02 Page 41 of 225 Microlab STAR User Manual 4 Design of the User Software 4 1 Overview The Microlab STAR User Software is the controlling software for the Microlab STAR It is a Windows based menu driven interface allowing the user to define deck layouts and methods and then to run the Microlab STAR The Microlab STAR User Software allows the user to program and run different methods for aspirating and dispensing liquids Note that each programmed method has to be validated by the user The deck layout is defined within the Deck Layout Editor whereas the aspiration and dispense steps are defined using the method editor An independent run control is used to run the methods programmed For ease of use there are two different options for programming In both cases a master or central editor is selected which can either be the method or the deck layout editor Whatever editor is started first is the master editor All other slave editors and the run control are then called from the master editor using easy access shortcut buttons If you make changes within one of the slave editors the master editor is automatically updated once you quit the slave editor This is illustrated below with the deck editor as master uh Ed Microlab STAR Edit Lb T Heb Heb Al NUM Run Control is Slave Sequence Edit
59. 0 Waste destination ml_star Waste This command ejects the tips to the sequence of the waste container ml_star Waste i e into the waste container We now need to get the program to keep count of the wells in the sequences adding one to the total so far each time a well is copied Create the following lines below the ml_star TipEject command but above the closing bracket Under variables double click sequence and select ml_star Source and Operator in the right frame Click OK Repeat this for the other sequences until you have written ml star Source ml_ star Target ml_ star Tip This tells the system to keep adding to the count of source plate wells to the count of target plate wells and to the count of tips When 96 wells and tips have been counted the counter will go back to zero thus ending the program in accordance with the while statement we invoked at the beginning The command automatically adds to the sequence counters the number of pipettings done so far The last lines in the method frame you should see are return The first bracket Y finishes the loop of the while statement return and the last bracket P ends the method TransferPlate Page 208 of 225 610766 02 Microlab STAR User Manual Finally your method should look like this aE Hamilton HSL Method Editor HSL_OnePlateToPlate File Edit View Inset Tools Win
60. 10766 02 Microlab STAR User Manual In the case of an error the process may be continued using the error handling procedure If for example a barcode of a carrier cannot be read a dialog window opens up Identify Error Error recover Code Description Continue Repeat Barcode Assigned recover son tee In this case there are 3 options to handle the error e Continue Continue without reading barcode again e Repeat Read barcode again e Barcode Enter barcode by hand A green dot stands for a tube where the barcode is read correctly A red dot stands for a tube where the barcode has not been read correctly An orange dot stands for a position with no tube not detected by the sensor After assigning a recovery option the Execute button is activated and the selected option is displayed in the lower part of the window Clicking on Execute causes the instrument to proceed lf an error occurs while using pipetting channels the displayed error dialog shows for every single channel its error state and its recovery options Different channels can have different errors 610766 02 Page 177 of 225 Microlab STAR User Manual For example in case of an LLD error such as no liquid in the container while aspirating a window similar to the following pops up Aspirate Error Channel selection Error recovery Code Channel 2 Description Channel 3 Mot enough liquid Channel 4
61. 195 197 200 201 203 205 Microtiter plates MTP 10 12 18 28 29 31 33 34 59 67 80 98 148 155 158 160 168 197 206 207 212 214 218 Options 1 3 29 Pause 29 110 194 195 Pipetting 3 6 14 20 21 29 30 47 67 81 82 84 85 87 92 94 106 112 116 135 137 141 142 144 152 153 155 157 158 168 185 236 Plate handling 6 184 Position 19 36 37 41 51 55 60 61 63 64 65 67 68 69 70 71 78 83 84 90 91 97 98 99 120 125 142 143 144 145 147 154 158 166 167 169 172 179 180 181 184 186 187 192 198 209 Power electrical 4 42 43 45 Probe 5 51 64 65 67 71 Rack 6 15 55 63 65 67 72 93 94 97 143 149 155 206 207 208 209 211 212 214 215 216 236 237 Run 5 6 10 15 29 42 49 50 51 52 04 57 67 76 84 85 91 109 110 111 126 137 140 148 154 158 189 190 191 192 194 195 196 203 204 Run abort 10 56 79 89 90 110 126 195 201 Safety 3 9 11 Sequence Editor 4 67 149 150 160 Sequences 4 6 50 51 55 57 67 68 69 70 71 72 73 74 78 81 82 83 84 85 86 87 89 90 91 93 95 97 98 99 106 131 134 135 140 141 142 143 144 145 147 149 150 151 153 154 155 157 158 160 164 166 167 168 169 170 171 172 173 174 179 180 181 184 185 186 187 192 Single steps 5 6 80 81 82 84 90 95 106 110 138 139 140 155 169 172 177 194 200
62. 237 238 Contamination and decontamination 3 11 15 20 Control PC 42 Deck 4 9 15 28 29 30 33 34 36 37 38 41 42 46 49 50 51 53 54 55 57 58 59 60 62 63 64 65 66 67 68 76 79 80 81 91 92 95 97 107 109 110 111 112 119 120 121 122 131 132 133 134 138 141 148 149 151 152 154 155 158 159 160 167 169 174 184 189 190 191 192 193 195 197 206 216 227 234 Deck layout 30 33 42 49 50 51 54 57 08 59 60 64 67 68 76 79 80 91 95 107 109 110 111 119 120 121 122 131 132 133 138 141 148 149 151 152 155 159 160 169 174 184 189 190 191 195 Dispensing 3 8 14 15 18 19 20 21 22 23 25 42 48 49 52 76 79 80 82 83 84 85 87 88 89 90 98 106 112 114 115 116 135 136 141 144 145 146 153 154 157 158 173 181 194 197 199 209 Disposables 3 33 Disposal of instrument 4 46 Errors 6 22 27 55 79 80 85 89 90 92 94 95 103 106 108 115 124 126 136 147 197 198 199 200 201 202 File structure 4 53 Firmware 4 11 42 98 Fuses 43 45 610766 02 Microlab STAR User Manual Graphical Method Editor 5 52 76 101 107 108 131 133 Gripper robotic hand 99 HSL Method Editor 5 7 51 52 53 76 96 102 107 108 111 131 197 Initialization 82 97 99 125 127 140 158 175 177 Installation 4 42 ISWAP internal Swivel Arm Platehandler 4 6 38 41 9
63. 3 Aspiration 3 8 14 15 16 17 18 19 20 21 22 23 25 26 27 42 49 52 59 76 77 79 80 82 83 84 85 87 88 89 90 97 98 106 112 114 115 116 135 136 141 144 145 146 153 154 157 158 168 172 173 181 194 199 200 Autoload 3 6 31 36 37 46 91 110 133 134 141 158 190 192 193 196 228 233 235 Automated Vacuum System AVS 4 5 32 38 39 40 41 119 120 121 122 124 125 126 127 128 129 130 Barcodes 6 9 31 32 36 53 55 103 141 184 187 190 197 198 203 204 Batch methods 5 106 Binding step return values 5 104 Biohazardous material 11 Carriers 6 15 16 18 29 31 32 33 34 36 59 60 61 62 81 85 92 97 98 99 132 133 141 147 148 149 154 155 158 169 174 183 184 185 186 187 189 190 192 196 198 216 217 218 Carriers temperature controlled 4 6 32 36 37 38 60 99 104 174 178 179 Channels pipetting 10 15 16 19 20 25 29 33 47 48 55 59 64 67 89 92 93 94 95 97 98 106 140 144 197 198 199 200 Cherry picking 6 158 Clot detection 3 27 Collision 59 Page 216 of 225 Configuration instrument 4 29 31 52 53 54 55 56 71 110 117 118 121 141 175 195 196 200 203 Configuration Editor 29 52 54 56 110 141 175 195 203 Container 6 10 11 14 15 16 19 20 21 23 29 34 36 40 55 59 64 67 83 84 88 97 98 145 147 199 207 209 211 212 215 234
64. 4 1 41 1 14 31 Potassium hydroxide a 3 os o D a Hes ee a Sodiumacetate 1 1 3 1 3 olill 41 3 4 al Sodiumborate 1 1 1i 1 3 o 1 4 1 41 4 a Sulfuric acid 1 75 2 4 1 f a 4 2 4fi1 4 41 4 s luring at Pt fa tt tt ft Pa ff lili ai Triethylamine Pa 4 fsalols o fila a1 fol a l4 toluene Cada aae a ge Se a ae ae ee Effects Key to codes in above table 1 no effect little or no noticeable change 2 Slight corrosion or discoloration 3 Moderate corrosion or other change in physikal properties or dimensions not recommended for continuous contact severe corrosion or physical change prolonged contact not recommended 0 No data 610766 02 Page 223 of 225 Microlab STAR User Manual Materials 1 4310 Steel POM Polyoxymethylene EPDM Ethylene propylene elastomer PP Polypropylene FPM Fluoroelastomer PTFE Polytetrafluorethylene NBR Acrylnitril butadiene rubber PVC Polyvinylchloride PE Polyethylene PVDF Polyvinylidenefluoride Storage Containers for Wash SI Silikon Liquids PEEK Polyetheretherketone 2 Chemical compatibility of higher resistance wash station Acetic acid 20 Pa 2 foals a a4 1 4 2 Aceticacia gaca 1 4 4 ia 1 fri a fa Acetone Pa 1 altel a i 1 alja Acetonitle 1 la 8 14 oo 8 a i lo Ammonium hydroxide 5 Pa a4 feta a kala 2 4 Chloroform Pa a fatal 1 fs 1 a a Deionizedwater tt tt at tf at at Dimethyl formamide Pa 2
65. 4 F Channel 5 Channel 6 Channel 7 Exclude Repeat Bottarn Ch E a Leanne Ar Move Up Assigned recover Red coloured channel 1 and channel 2 failed to aspirate To each of these two channels a recovery option has to be assigned Only if all channels have the same error is the chosen recovery option assigned to all channels simultaneously selecting channel 1 shows the error description and the available recovery options for this channel buttons Exclude Repeat etc Invoke one of these options by clicking the appropriate button Your selection is displayed in the lower part of the window In this case there are 5 options to handle the error e Exclude The channel is excluded no more aspirate dispense etc with this channel until next TipPickUp e Repeat Repeat aspirate command e Bottom Aspirate from bottom of container without LLD e Air Aspirate air e Move Up Channel is moved up to dispense liquid into the container After this action aspiration can be repeated Note that the Continue button is disabled This prevents any later dispense with insufficient volume Repeat the same procedure for channel 2 Note that the error and the associated recovery options may differ from those for channel 1 When the last channel is processed the Execute button becomes active and the system can proceed In any case the method can be aborted without further recovery options Page 178 of 225 610766 02
66. 6 13 12 07 10 58 55 Microlab STAR tubes 3 tips 13 12 07 10 59 19 Microlab STAR Atipcamerstips_ s gt comet Heb Select a run in the upper half to get a list of labware processed during the run Double click on the plate for which a barcode file is to be generated Another window opens up displaying a table with the pipetting information of the plate in a convenient form Bat Barcode File Filter plate 0 Iofs E File View Options Window Help 2 E x De SIIB wad 12 hn A l AT Barcode F ile Filter Page 1 of 2 Pipetting Protocol Example hsl Fun Interrupted plate cc 13 12 01 1454 54 Position Pipetting Information Nt i ec pt Ui pt i 2 O O SS Ee Sed AOS tt Uc Aan Blot zed ee lg The table may be printed from the menu To start the AT Filter automatically at the end of a method see the example in chapter 13 7 610766 02 Page 183 of 225 Microlab STAR User Manual 17 Reference Guide Defining Labware 17 1 The Labware Editor If your labware is not pre defined you can define custom racks and containers using the laoware editor Such custom labware can be used like any other pre defined labware object from the library The idea is that the labware object is a description of the real physical labware To start the Labware editor from the deck editor 1 select Add Labware from the Edit menu or from the popup menu and click the Lab Editor button or 2 select the Lab
67. 66 02 Page 155 of 225 Microlab STAR User Manual select the sequence ML_STAR WashStations holding all individual wash modules Click OK Aspirate from the Samples sequence and dispense into the Plate sequence with the settings as described in the example Tubes ToPLate Eject the needles with the SMART Step iil Needle Eject Edit Instrument short name Needle sequence ML_5 TAR Wash Stations al MW Start wash Channel settings Error settings Cancel Help Again select the sequence ML_STAR WashStations and check the checkbox Start wash The SMART Steps needle pick up and needle eject will handle the washing within the alternating three wash modules automatically Finally the carriers may be unloaded Page 156 of 225 610766 02 Microlab STAR User Manual 13 8 A General Example for User In and Outputs Sample Tracking Sequence Manipulation Tip Counter and TCC Use The following example gives an idea of the additional functionality of the Microlab STAR user software Create the following deck layout holding a tip carrier TIP_CAR_480 with tip racks at least two racks 3 tube carriers car32_cup12x 5 rck for 32 tubes anda temperature controlled carrier car_tcc_1 tml named TempCarrier with a standard nunc plate nun_96_fl_I rck named plate Save the layout under the name Example do not use a name longer than 20 characters
68. 66 02 Page 43 of 225 Microlab STAR User Manual 4 2 1 Save As or Saving Methods and Deck Layouts under Different Names Due to the n n relation between deck layouts and methods the Save as option needs to be used in the following way To save a method under a different name still referring to the old deck layout e Select Save As in the method editor and save the old method A under a new name B The new method B is still linked to the old deck layout A To save a method and the corresponding deck layout under a new name e Start the Method Editor as the first opened master editor e Select Save As to save method A under the new name method B e Start the Deck Editor to save deck layout A under the new name deck layout B Close the Deck Layout Editor e Link method B to deck layout B Methods gt Instruments Save method B 4 3 Structure of the User Software The User Software consists of a set of modules and tools programs which work together These modules and tools are grouped into a general instrument independent and a specific instrument dependent group Given that structure a system with different HAMILTON instruments even non HAMILTON instruments suitably adapted can be controlled and operated in the same manner in the same environment and at the same time The general modules get all the specific information they need about instruments such as pipetting steps or information about the probe
69. 75 mm 150 SMP CAR 32 Eppendorf tubes 1 5 ml 50 SMP CAR EPIL Eppendorf tubes 0 5 ml 50 SMP CAR EPIS 96 well PCR plate 200ul well 50 PLT CAR L5PCR 96 well flat bottom microplate 75 PLT CAR L5MD 384 well flat bottom microplate 50 PLT CAR L5MD 96 deepwell microplate archive _ 150 _PLT CAR LSAC Once the liquid surface is detected an additional immersion depth typically 2mm is reached to prevent the aspiration of air and aspiration starts The tip follows the falling liquid level if so specified according to the volume aspirated Then the tip leaves the liquid slowly and heads for the vessel to dispense into Finally to prevent droplet formation a variable amount of transport air is aspirated After an aspiration step the situation within the tip looks like this Channel Blowout air optional may reach into channel Liquid Transport air optional The situation within the tip and the channel after aspiration 610766 02 Page 15 of 225 Microlab STAR User Manual 2 2 3 Dispense The transport air that was aspirated at the end of the aspiration step is first dispensed with the tip still in the air Dispensing of the liquid may now occur in two different modes to the liquid surface or in a jet Dispense in Jet Dispense to Surface I To ensure that the specified accuracy is achieved volumes below 20 ul should always be dispensed to a liquid surface For dispensing to a liquid surface use cLLD to detect the positi
70. 766 02 Page 165 of 225 Microlab STAR User Manual 71SWAPEXAMPLE lay Hamilton Deck Layout Editor ae 5 x File Edit View Tools Help OSa S Flagler aja ae il bball age l T FlateAtExi reagent Flate12 lated2 ae ieee aie a iil a age 7 For Help press Fl ix 950 000 y 454 000 EA Create independent sequences for pipetting steps and transport steps using the sequence editor Both the transport and pipetting sequences span all 3 plates The first step of the method is a simple aliquot step to transfer reagent to the plates located at plate carrier 1 using the SMART Step Pipette Page 166 of 225 610766 02 Microlab STAR User Manual einem H Pipette with Tips from ML STAR Smart Steps Instrument short name MLLSTAR Aliquot procedure Volume per well ul 20 Aspirate sequence WL_STAR ReagentSequence Dispense sequence ML_STAR PipettingSequencePlatel Tas controlling sequence over following sequences ML_STAR TransportSequencePlatel Tas tee ounterT used as loop counter variable Get Plate on ML_ TAR Movement type 0 To carrier Transport mode 0 Plate only Sequence ML_STAR TransportsequencePlatel Tos Sequence counting 1 Automatic Return value count 3 Move Plate on ML_STAR sequence ML_STAR PlateAtExternalbC Scanner Return value count 3 Place Plate on ML_STAR Movement type 0 To carrier Transport mode 0 Plate only Sequence
71. 9 184 187 Labware 4 6 16 18 28 33 50 51 53 57 59 60 61 62 63 64 65 66 67 68 69 70 71 91 120 121 149 155 184 189 190 192 204 206 207 208 216 217 235 Labware Editor 6 206 216 Libraries 5 53 62 101 102 103 108 161 164 166 167 174 179 184 206 Life cycle of instrument 46 Liquid Classes 3 5 9 20 21 22 23 25 27 52 81 112 114 116 117 118 135 144 146 Liquid Editor 5 21 52 112 113 116 117 Liquid level detection LLD 9 10 15 16 17 18 22 23 27 88 97 98 136 144 145 154 158 199 209 218 Liquids 3 5 8 9 10 11 12 14 15 16 17 18 19 20 21 22 23 25 27 35 36 37 40 47 48 52 55 67 81 83 88 89 97 98 110 112 113 114 115 116 117 118 135 144 145 146 148 153 158 169 197 199 201 203 209 Loading and unloading 6 28 31 37 38 59 80 91 92 110 133 135 147 154 157 158 167 168 183 189 190 192 193 196 197 Maintenance 4 10 11 46 Method linking 5 111 Methods 5 6 7 9 26 38 41 42 48 49 90 51 52 53 54 56 57 59 66 67 76 77 78 79 80 81 82 89 91 93 96 99 100 101 102 103 104 106 107 108 109 110 111 112 119 121 122 123 126 128 130 131 132 133 134 137 138 139 140 141 147 148 152 154 155 157 158 159 160 161 610766 02 162 163 165 169 170 174 175 177 178 180 183 185 186 189 190 191 192 193 194
72. E o O EE AE EAN AE AEE A AEE E E E 117 13 2 Create a Method to Copy from Plate to Plate Using SMART Steps seesees 117 13 3 Create a Method to Copy from Plate to Plate with Single Steps sessecccosssssseesoosssoe 124 13 4 Create a Method to Copy from Tubes to Plates Using SMART Steps 000 134 13 5 A Method to Pipette Aliquots Using SMART StepS sseeccosssssseeccoosssesseccocsssssseeesosssoe 140 13 6 Cherry Picking or How to Change a Sequence Within a Method Usine SMART SEPS a a a a aea aE a 142 610766 02 Page 5 of 225 Microlab STAR User Manual 13 7 A Method Using Steel Needles Single and SMART Steps Including Sample Reduction seses si cdecsiavscdecssssccsaceveceocssicctuecosacecssscceussacatessscseatesessheteassase evs 153 13 8 A General Example for User In and Outputs Sample Tracking Sequence Manipulation Tip Counter and TCC Use ccccsssssssssssccccccccecsssssccccoees 157 13 9 Using the SWAP robotic plate handler oooosssssseeseceseeccoocsssssssssssecsesesesoossssssssssssessee 165 TA Loadino the Microlab S TAR rasieren EE E aes 169 MAA Manual a iison E E a E EE 169 EY SUMMING the Microlab STAR srno E TEE EA 171 15 1 Running a Sample Method with the Instrument ccccccssssssssccccccccsssssssscccesseeeees 171 152 R n Simulations osne e A E E 175 153 R time Error Fi arid in aisis a aaa Eaa a AaS 176 15 4 Walk Away Predefined Error H
73. If you loop over more than one sequence the shortest sequence is will be taken as the relevant one Page 128 of 225 610766 02 Microlab STAR User Manual Under the ml_ star toolbar drag TipPickUp to the position between the two loop statements Step TipPickup Edit Sequence settings SEQUENCE Sequence counting 1 Automatic Channel settings Error settings A dialog box appears Choose sequence ml_ star Tips and sequence counting Automatic Sequence counting automatic means that after pick up takes place from the first eight sequence tip positions the tip sequence is automatically incremented Sequence counting manually means that the sequence used in this step will not be incremented automatically during the next tip pick up process the positions within the tip rack already used will be used again Therefore we select Automatic to increment the sequence automatically by the number of channels used for the tip pick up Next time tip pick up will start with the next unused 8 positions in the same tip rack Click OK 610766 02 Page 129 of 225 Microlab STAR User Manual Now drag Aspirate to the line below the tip pick up step A dialog box appears Aspirate New Sequence Sequence counting ML_STAR Source fi Automatic Aspirate mode pl Individual volumes IO Aspiration Tip type Dispense mode Standard volume tip without filter
74. L The general syntax used in HSL is the C C notation Instrument steps are shown with a gray background For safety reasons it is not possible to change or type in these commands directly Double clicking an instrument step displays a step dialog where you can change specific parameters To see all step parameters in the Method View use View All Stepdata in the View menu The Toolbox Window on the left offers various elements that can be used to build the method grouped by the following bars Bar name Contents ML_STAR Each device defined in the method shows its own instrument bar as example of a device containing all the steps the instrument can execute language Contains the common HSL language elements such as if else loop function call etc library function A set of standard library functions such as mathematical functions trace functions string manipulation functions etc variables Different variable types All these elements can be used by dragging them from the appropriate bar and dropping them into the Method View or by using the nsert menu Alternatively language elements library functions and variables but not instrument steps can also be typed directly into the Method View When an instrument step is inserted the appropriate step dialog is displayed so you can define the specific parameters needed After writing a method invoke Syntax Check from the Too s menu to check your method The Syntax Check Wind
75. Microlab STAR User Manual Microlab STAR User Manual for User Software 3 0 including Options and Accessories HAMILTON Bonaduz AG 610766 02 Page 1 of 225 Microlab STAR User Manual Page 2 of 225 610766 02 Microlab STAR User Manual TABLE OF CONTENTS L General Informan osin cous desc guasencensedelccsuasaneasses eee azeisensens aE ana 7 1 1 About this Mantal aaa aaaea aaa aa aaae a E aaa a aa aaaea aa aaaea 7 1 2 Additional Microlab STAR Manuals sissicceccscbessesesdnsecssecasesosnscdadscobsaeaessseacsdessosvisaccducsscnvewsbeses 7 1 3 Intended Use of the Microlab STAR viirin orttirsa eose ea iaa aa aa a 7 kae SOMO EAGION E E E N E EA EEE 8 ES SACI ii a E ae R rae a a a n aa a 8 bal GreneralePrecati Ons unan N E E E 8 Lo HelectnCal Satcty PreCaull OMS vette cect onsseratgia a SonasnaSeasesadconsgnsSadwanch ea a E 9 L3 Biohazard Precautions asjesisgauseustcneipi este scecesaussguouvscees seeietenan a ni apeouds A RN awieaaes 9 Wad Computer Precautions ets asceateietstoeaehan Gk satan E a banteneteietet ae vas cah sha seatan nes eeateceutenl 9 LG Warranty Statement asrasa oaa a a Eaa AE EA TAEAE A 10 1 7 Customer SCLV ICG sosire aea aA E a aaa a a E aE EEE Naaa 11 Z BNE ATE OF PIDCUING coudewedewccisasetenineuuseicesacasdenesvocdascae texas ENE ede em 12 2 1 The Air Displacement Pipetting Principle sssessecccocssssssccecosssssseecoosssssseccsoosssssseesosssssssee 12 2 2 From Aspiration to Dispensing
76. Pick trc using the library function TrcTrace8 to inform the user Page 150 of 225 610766 02 Microlab STAR User Manual This function adds a text of eight variables to the trace file Se TrcTrace of HSLTrcLib Edit Library name C Programme HAMILTONSLibrarySHSLT reLib he Warable Function name TreTraces P Function parameters e Ta posi C A E C E e C Cancel Help After the file has been analyzed and the Hit sequence has been generated the loading of the sequences carriers to the deck may start The load SMART Step looks like this Sequence Read tp Tip counter Start pos Wo of pos 1 Redueible a gC ne E Moo R e O a m startot ol O CU OT NOTE The sequence Hits that has been freshly created and which has a current position of 0 because it has reached its end position is automatically set to 1 by the loading step If there is no entry in the field Start Pos then the current position of the sequence is set to the first position 610766 02 Page 151 of 225 Microlab STAR User Manual The pipetting is then performed by the SMART Step pipette The important sequence settings are shown in the next screen i Aspirate sequence is controlling C Dispense sequence is controlling Aspirate sequence ML_STAR Hits C Reload 4 Reducible by user f Not reload Dispense sequence MLS TAR T arget ka C Reload e
77. R User Manual Waste Container N s The waste is collected in the closed system waste container with liquid level monitoring to prevent overflows The waste container with a volume of 10 litres should preferably be placed below the level of the instrument The waste level sensor has to be connected with the controller F a The AVS and its components are to be installed by a Hamilton service engineer 3 3 6 iSWAP The ISWAP internal Swivel Arm Plate Handler transports microplates covers of microplates archive plates or filter plates used for the vacuum box to and from positions on the deck of the Microlab STAR IVD The plates can be placed in landscape or portrait orientation and rotated 180 degrees In addition the ISWAP is able to load and unload plates to and from a plate stacker on the left side of the instrument outside the working area with some restrictions this can also be done on the right side Like Microlab STAR s pipetting channels iSWAP has a travelling height of 145 mm above the deck 245 mm above the origin The iSWAP is mounted on the pipetting arm and parked below the cover of the pipetting arm In this position it does not affect the movement of the pipetting channels ISWAP robotic hand ISWAP can be chosen by the customer as an ordering option but can also be installed later by a service technician at the customer s request 610766 02 Page 35 of 225 Microlab STAR User Manual 3 4
78. Reducible by user Multiple use I Aspirate all LLD Advanced pense mae LLD Advanced i Jet C Surface Liguid liquid class Water Standard olume Water Disnenselet Finish Cancel Help The aspiration sequence the hits is controlling here and not reloadable The target sequence is not reloadable either The rest of the settings is very similar to the previous example OnePlatel oPlatePipette Finally the unloading takes place in the usual way You will find additional examples involving for instance the reading of a comma separated file in MTP map format HitPick_ CSV in your demo methods directory Page 152 of 225 610766 02 Microlab STAR User Manual 13 7 A Method Using Steel Needles Single and SMART Steps Including Sample Reduction The following method transfers liquid from tubes to a plate using steel needles The aspirations and dispenses are done with single steps and for the washing of needles the SMART Steps are used A reduction of the number of samples is possible The method uses the following deck layout A plate and a set of four tube carriers of 24 tubes is used Add the standard plate carrier PLT CAR_L5MD Add a Nunc plate nun_96_fl_I rck and name it Plate as well as the tube carriers car24_cup15x100 rck as decribed previously In addition a wash station is added to the rightmost position on the deck Select the carrier for the wash station car_wa
79. S TAR Source Reload Reducible by user f Not reload F Aspirate all LLD Advanced Liquid liquid class Water Standard olume Water Dispense et z Dispense sequence is controlling Dispense sequence MLS TAR T arget C Reload M Reducible by user f Multiple use Dispense mode LLO Advanced tf Jet Surface Cancel Help Choose Standard Pipette Procedure and Simple as a pipetting mode because it s a simple transfer from plate to plate The volume to be transfered is 50 ul the residual volume is 0 Select the tip sequence ML_STAR Tips from the dropdown field The tip handling chosen here is to take new tips for each sample which is for 50 ul and standard tips the same as after each sample Now select the aspiration sequence as the controlling sequence Even if both sequences have the same length initially it makes sense to choose the aspiration sequence as the controlling one Remember that on loading this sequence the sequence may be reduced to less than 96 positions Then only the current number of wells is transfered to the target plate Both aspiration and dispense sequence are set as not reloadable The dispense mode is to dispense in a jet because initially the target plate is assumed to be empty The liquid class used in this example is water 610766 02 Page 121 of 225 Microlab STAR User Manual On aspiration you may use capacit
80. TRUE FALSE MiStarCarlsLoadable 0 0 or 1 The carrier can be loaded and unloaded A wash station for example is not loadable 1 TRUE 0 FALSE Page 198 of 225 610766 02 Microlab STAR User Manual MiStarCarlsAutoLoad 0 0 or 1 The carrier can be loaded and unloaded by the autoload 1 TRUE FALSE MiStarCarPosAreRecognizable 0 Oor 1 The presence of the elements loaded on the carrier is detected e g the containers on a sample carrier are detectable 1 TRUE FALSE MiStarCarNoReadBarcode 0 Oor 1 1 Don t read barcode 0 read barcode Notes e The position of the carrier barcode is not defined in the properties because this position is fixed e The barcode positions are ordered at regular intervals one measurement for one carrier e Onacarrier all barcodes must have the same orientation except the carrier barcode one value for one carrier e The barcode read window is the same for all positions on a carrier One window should not overlap the next window otherwise in some cases the barcode cannot be assigned to the correct position e lf acarrier is loadable with auto load MIStarCarlsAutoLoad the loadable property MiStarCarlsLoadable value must be set too e At edit time in the load step the user can only select carriers templates with underlying carriers having the property MlStarCarlsAutoLoad e Atruntime if MlStarCarlsLoadable and MiStarCarlsAutoLoad are
81. Technical Support department at HAMILTON Bonaduz AG will be available by phone fax or e mail to deal with your queries Here is their address phone fax and e mail Europe Africa and Asia HAMILTON Bonaduz AG Phone 41 81 641 6060 Technical Support Fax 41 81 641 6070 P O Box 26 CH 7402 Bonaduz Switzerland E mail itechsupport hamilton ch Americas Far East and Pacific Rim HAMILTON Company Toll Free 800 648 5950 P O Box 10030 Phone 775 858 3000 Fax 775 856 7259 Reno NV 89520 0012 USA E mail tech hamiltoncompany com 610766 02 Page 11 of 225 Microlab STAR User Manual 2 The Art of Pipetting In this chapter the process of pipetting with the Microlab STAR is described Pipetting means transfer of small quantities of liquid from one container to another A pipetting operation is achieved by aspirating drawing liquid from a source container then transferring and dispensing dropping it into a target container 2 1 The Air Displacement Pipetting Principle The Microlab STAR is based on the air displacement pipetting principle comparable to the functioning of hand pipettes Air displacement means that the liquid is aspirated into and dispensed from a disposable tip or needle by the movement of a plunger Between the plunger and the liquid surface is air No system liquid of any kind is involved in the Microlab STAR CO RE stands for Outer Release Sleeve Pressure Sleeve O Ring p Coupling Shoulde
82. _O002 Return value count 3 1 UnloadCarrier on ML_STAR MULL Rack type plt_carlSime_O001 Return value count 3 The Copy from Plate to Plate Method Using single steps 610766 02 Page 125 of 225 Microlab STAR User Manual Let s start to construct this method Under the ML_STAR toolbar drag Initialize to the main window A window pops up Initialize New Sequence settings Use default waste 1 Un Waste destinatior Sequence counting ML_s TAR waste T 01 Manually Always initialize 1 On Channel settings Error settings ee Click OK to initialize the instrument in the first step The waste sequence is selected to eject tips from the channel during initialization Always use manual Sequence counting for initialization see below Set the Always initialize switch to On to make sure the instrument is being initialized prior to each run Throughout all single steps the channel pattern to be used may be specified manually Clicking on Channel Settings opens the following dialog Channel Settings Channel pattern E Channel selection as varnable Channel selection Channel variable Channel use fi All sequence positl Cancel Help You may either deselect specific channels manually by clicking on the check boxes or check the box selection as variable Then the channel pattern can be defined as a string variable
83. _star directory Click OK in the add labware dialog Add Labware to DeckStte EES Hame M pPlateL arrier Type rol staryplt_car_lSrnd tml us Lab Editor OK Cancel The carrier is now added to the deck layout i Unbenannt Hamilton Deck Layout Editor OF File Edit View Tools Help Djela S Flagler Ala e Edit Method Aun Method 5 30 35 40 45 For Help press F1 a 669 000 Y The next step is to add a plate to this carrier The corresponding labware plates tip racks etc is sorted by manufacturer and stored within the Labware directory Standard plates from almost every major manufacturer are available Nunc Falcon Greiner etc 610766 02 Page 53 of 225 Microlab STAR User Manual Microplates tip racks etc are labware items and generally called racks The defining files have the extension rck To add a plate to the carrier double click on the desired carrier position Again the add labware dialog box opens Select the Nun_96_FI_L rck 96 well flat bottom Nunc plate in landscape orientation from the labware nunc directory Give a name to the plate e g MyPlate i Unbenannt Hamilton Deck Layout Editor OF x File Edit View Tools Help oela S wl asa Ala e Edit Method Aun Method 15 20 5 30 35 40 45 For Help press F1 E aos 000 Y 2 The plate is added to the site Plates or labware may be moved betw
84. a bifurcation for programming parallel processes For a detailed description of the input dialogs that correspond to these icons refer to the on line help 8 2 Using Microlab STAR Specific Steps To use instrument specific steps e g aspirate dispense etc the method has to be linked to an existing deck layout first Use nstruments in menu Method to open the following dialog Instruments x Short Instr eal Deck Layout File Low Level aee Microlab STAR ML_STAR Visible The instruments dialog Click on to browse for the deck layout you wish to write a method for Select the layout from the file list and click Open Instruments M Microlab STAR ML_STAR test lay Visible If the selected deck layout corresponds to a Microlab STAR the check box is enabled Click OK to link method and deck layout Keep the Smart Steps check box selected Note that multiple deck layouts may be linked to one method to enable programming of different instruments within one method Once you have written a method using instrument specific steps the button vanishes Now the deck layout can no longer be separated from the method until all instrument specific steps are deleted again 610766 02 Page 69 of 225 Microlab STAR User Manual NOTE Only after the method has been linked to a deck layout are the instrument specitic commands loaded into the method editor Linking the deck of another instrume
85. ains at least one barcode file At_barco nij plus the two register files containing information about all runs performed so far for a description of reg files refer to the Microlab AT user manual a At_bareo 001 a At_bareo 002 a At_bareo 003 a At_bareo 004 a At_barco 005 Ei Register int FRegneu int The barcode files can be directly used to couple the Microlab STAR and Microlab FAME Now start the filter tool manually by selecting START gt Programs gt Hamilton gt Micolab STAR gt AT Barcode Filter lel Click onthe icon Page 182 of 225 610766 02 Microlab STAR User Manual A window opens up showing the available runs E Create an AT Barcode File E Available Auns Sample Tracking hal 13 12 01 09 45 45 13 12 01 09 47 15 Mo Error C PrograrnimesH4 SingleStepE sample hal 13 12 01 09 53 08 13 12 01 09 54 13 No Error C Programme HA SingleStepE sample hel 13 12 01 09 57 51 13 12 01 10 00 27 Mo Error C Frogramme HA Example hs 13 12 01 10 17 25 13 12 01 10 19 36 Mo Error C Frogramme HA Example hs 13 12 07 10 21 18 13 12 07 10 24 47 Mo Error LC Prograrnime sH 4 E sample hel 13 12 07 10 58 34 13 12 01 10 59 19 Fatal Error L Programine sHAgag Elements for the selected Aur Loading Time Full Element Name plate CE 13 12 01 10 58 45 Microlab ST AR4T empCarrierp tubes 12345676 13 12 07 10 58 48 Microlab STAR AtUBes1 tubes 12345676 13 12 07 10 58 51 Microlab STAR Atubes2 tubes 1234567
86. ance based LLD Click on the LLD button Aspirate Liquid Level Detection LLD Capacitive LLD Pressure LLD T Enable pressure LLD Sensitivity f Ver high Middle fe Ver high ft fiddle High C Low e High CE Low Submerge depth mrm Aspirate height from bottom rrn Mas height difference rrm Cancel Help Set the settings according to this example Click OK On dispense switch off the LLD and dispense to a fixed height 2 mm from bottom Dispense Liquid Level Detection LLD Capacitive LLO P Enable capacitive LL Sensktivity i Wen f Figh ai Mirade i H i ah i Lig fad Detail settings Submerge depth mrm Dispense height from bottom mrm Poo B Cancel Help Accept the defaults for the settings under Advanced and for the error settings Click OK Click Finish in the SMART Step dialog A window appears showing a summary of your settings Click OK Page 122 of 225 610766 02 Microlab STAR User Manual Finally drag the SMART Step Unload to the line below the pipette step iii Unload Edit Remove Click on Add all Sequences to add all sequences to the unload step Click OK Within the method editor click File gt Save to store your method Your first method is ready to go See the chapter about running the STAR to see how to run your method 610766 02 Page 123 of 225 Microlab STAR User Manual 13 3 Create a Method to Copy from Plate to Plate with Single Steps
87. and thus the decision which formula is used to calculate the reference z height from the labware data is done automatically by selecting the hidden grid for the rack Thus the question arises how to define the hidden grid for a rack i e how to define which hidden grid or carrier the rack should snap onto For a rack on top of a carrier this is done by selecting the appropriate switch snap to base in the carrier definition For a tube rack which is placed directly on the deck it snaps directly into the 1 track grid the grid of the tracks automatically assumes a rack based snap on When placing a plate directly on to the deck the user has to input the x y z coordinates of the reference well directly 610766 02 Page 203 of 225 Microlab STAR User Manual 18 Reference Guide Examples Using the HSL Method Editor All examples shown in the following sections are identical to those in the previous sections except that they are written using the HSL Method Editor Therefore the previously created deck layouts can be used here as well Open a deck layout from the graphical method editor examples section and save it under a different name We have chosen the same file names as those for the other sample methods but with the notation HSL_ prefixed to the name All these methods are also available from the DemoMethods_ MLSTAR directory To link the Deck Layout Editor permanently to the HSL Method Editor the value of the regist
88. and consumable products e g carriers racks tips steel needles and waste containers are warranted to be free of defects in material and workmanship at the time of delivery only The above warranty shall not apply if e the product has not been operated in accordance with the user manual e the product is not regularly and correctly maintained e the product is not maintained repaired or modified by a HAMILTON authorized representative or user e parts other than original HAMILTON parts are used except for liquid containers such as microtiter plates and tubes e the product or parts thereof have been altered without written authorization from HAMILTON Bonaduz AG e the product is not returned properly packed in the original HAMILTON packaging HAMILTON reserves the right to refuse to accept any product that has been used with radioactive or microbiological substances or any other material that may be deemed hazardous to employees of HAMILTON Such a product has to be properly decontaminated and marked HAMILTON endeavours to provide prompt and satisfactory service Page 10 of 225 610766 02 Microlab STAR User Manual 1 7 Customer Service Customer service will be provided in the first instance by the network of HAMILTON representatives In the event of any problem experienced with your Microlab STAR the first recourse is your local HAMILTON representative For further problems requiring hardware or software expertise the
89. andling cccscsccccssssssssssccccccccssssssscccssseeeees 179 16 The Microlab AT Barcode File Filter sssssssssssoooosssssssssssseseesecocoooosssssssssssccesessseoososss 182 17 Reference Guide Defining Labware s sssssssssossssccoosseccccossssecoccssecccosssseccosssscecosssseesoo 184 L The Teabware Editores sequsucssssdecsepnesseaues consatoveesecs 184 2 SEV OES OL AD WAL e wisssscteosionsce tests terssoseGoesnartes cevsuGs baaaustucasesngoaesasstsecsusugitevssadescsunecanessaetetcauas 184 By Rectangular Racks ands P ates mrke iene aie e E iea E AEE 184 L722 COMAE asana R E E a N E 185 17 23 Cire ular Rae kSnnen a a a e a R E tattey 185 17 3 Example Defining a Rectangular Rack with Containers ssssssssecceceecoooossssssssssseseee 185 17 3 1 IDehinimG a Comain eina E E a 185 I Detinine a Rectansular Custom Rac kasuse a a tetera tein dat eoaaud 188 L733 Defining a Carer Chempl ate ij1 secs coseiieids eat ai a E uae 193 17 4 System Flags for Labware Properties ccccsscccssssssssssssccccsccsssscssscccsssccesscsssccosssceeees 198 17 4 1 SU IC CIC as E EA EE suchas O E E A E S T A E A E 198 17 4 2 Information for Handling the Autoload Unit 2 0 0 ccccssssssessseeeecceeeeeeeeeeaaeaassnssseeseeeeess 198 17 4 3 Information TOL o pecial UMIS sa les athe tose tends A a elsentaenacs wee bteuinauiasees 199 17 5 Z Positions of Carriers Racks and Containers cccccccsscsssccssccssccssccssccssccssccs
90. annot go any further down Clearance heights of rack and container Page 202 of 225 610766 02 Microlab STAR User Manual Regarding the z position two different cases of rack placement on the deck can be distinguished in the Microlab Star 1 A container based rack is placed with the container bottom directly on the carrier e g the microplates on a plate carrier Zinick thickness of container bottom Z eaiier Leck 100mm The reference position Zo is marked with the Here the reference height is calculated from Zo LoecktZearrier Zahick ignoring all other terms Zaeck is a fixed quantity Znick is defined in the labware and Zoarrier is defined in the carrier template definition 2 A rack based rack is placed with the frame on the instrument deck e g a tube rack where Zearier 0 because the tube rack is used directly as a carrier 4 Zihick thickness of container bottom Zpasediff distance from base of rack to base of container Zaner Liek 100mm The reference position Zo is marked with the Here the reference height is calculated from Lo Loeckt Zearriert Zbaseditt Zthick Again Zueck IS a fixed quantity Zihick and Zpaseaifp are defined in the labware and Zearrier IS defined in the carrier definition Note that here we have one more term in the equation compared to the previous case Selection between the two types of racks container based rack or rack based rack
91. apacitive liquid level detection a sufficient conductive coupling of carrier and labware tubes or microplates is crucial For non conductive liquids or in case of an insufficient electrical coupling between container bottom and carrier pressure LLD should be used Pressure LLD only works with new and empty tips for the aspiration of liquids The suitable settings depend on the tip size and on the type of liquid 300 ul channel 1 Very High Standard Low boiling point low viscosity 2 High Low Low boiling point low viscosity 3 Medium Standard Water or higher viscosity 4 Low Low Water or higher viscosity pLLD Setting Sensitivity Level Tip Liquid Liquid O O 1 Very High Standard Low boiling point low viscosity 2 High High Low boiling point low viscosity 3 Medium Standard Water or higher viscosity 4 Low High Water or higher viscosity In the case of aspiration from foaming liquids capacitive liquid level detection in particular may not detect the surface properly As an alternative try pressure LLD or a combination of both If you are using a combination of both LLD types the maximum height difference between the two independent LLDs can be used as a parameter Page 14 of 225 610766 02 Microlab STAR User Manual The following table gives dead volumes for both pressure and capacitance based liquid level detection in various containers Labware MVrnin ul Carrier Tubes 16 mm x 100 mm 200 SMP CAR 24 Tubes 12 mm x
92. arlelelarlarlerarlelelalarlavlalerlatalale axalelalelexalelealalarlerlarlelalalarlavtalelarlalale axalelalelerxalelelarlarlerarlealalalalavlalalatlalale axalelalelerxalalelalalearerlelelarlarlavtalarlarlalale alalalealealelerlelelalalelelelelalalalelalalerlala aralelaleleralelelalalerlarlelalalarlavlealalarlalale alalalelelelelelelalatlelelalelalalarlelalalelale Source sequence is cut to match length of target sequence stops when plate is full prepare plate for assay Case 3 L Suisp gt L Sasp Control ASP le le E je e e le le E le ej e E fe e e e fe e e e e e E fe e e e e le e e e e e CIC Se e E e e e E e e Target sequence is cut to match length of source sequence stops when tubes are processed how many samples today If L Sasp gt 96 plate is filled again Case 4 L Suisp gt L Sasp Control DISP 610766 02 Page 75 of 225 Microlab STAR User Manual OO0O00000 Source sequence is repeated to match length of target sequence stops when plate is full Distribute Buffer Reagents Also used for aliquoting Sequence handling in Pipetting Mode Pooling and Replica In these two cases the aspiration sequence should be selected as controlling Sequence handling for Aliquot Procedure In the case of aliquoting the longer dispense sequence is to be selected as controlling Page 76 of 225 610766 02 Microlab STAR User Manual For th
93. ash parameters see the section on the wash station 8 2 1 4 SMART Steps for Picking up and Releasing Tips Two further SMART steps allow you to pick up tips from a rack and then dispose of them in the tip waste after use Here is the dialog for the pick up tips step HM Tip Pick Up New Instrument short name Tip sequence x M Reducible by user Tip counter Tip counter Write tip counter Channel settings Error settings Cancel Help The dialog asks you to specify a tip Sequence You also have the option of specifying a tip counter 610766 02 Page 83 of 225 Microlab STAR User Manual Here is the dialog for the tip eject step E Tip Eject New Channel settings Error settings Cancel Help For channel settings and error settings see the description of previous SMART steps 8 2 2 Single Steps Now look at the Toolbox Window You will find that for each deck layout or instrument you selected there is now an additional bar containing the instrument specific steps ML_STAR H BE General a des ML_STAR il Liquid handling al i Preparation M Initialize wi LoadCarrier ba Unioadlarrier cee a Set Carrier Tempe T E Get Carer Tempi Z B Calibrate B Move Tl oPosition Ae Firmware Command ar r ML_STAR Smart Steps Method Editor with instrument specific steps in ML STAR Smart Steps two different views Pag
94. at is identified from the extension of the worklist s file name The editor supports Microsoft Excel xls Microsoft Access Database mdb Text Files txt csv tab asc No other extensions are supported 8 8 2 Worklist Handling with Microsoft Excel You can specify a subset of the available data when you first open a Microsoft Excel workbook In a workbook file you can open a single worksheet a named range anywhere in the workbook or an unnamed range in a single worksheet 610766 02 Page 93 of 225 Microlab STAR User Manual The following table lists the conventions for these settings Entire worksheet ina workbook file Specify the sheet as sheetname where sheetname is the name of the worksheet Important You must end the worksheet name with a dollar sign Named range of cells in a worksheet or workbook file Specify the named range as NamedRange where NamedRange is the name you assigned to the range in Microsoft Excel Important You must name the range in Microsoft Excel before attempting to open it Unnamed range of cells in a single worksheet in a Specify the sheet you want to open as sheetname and the range as FirstCelllnRange LastCelllnRange For example to access cells A1 through Z256 in a worksheet workbook file called Sales you would specify Sales A1 2256 Specify the sheet as sheetname where sheetname is the name of the worksheet Entire worksheet in a workbook file Importan
95. atePipette lay Page 136 of 225 610766 02 Microlab STAR User Manual Click on the Edit Method button to open up the Method Editor From the editor s Method menu select Instruments to link the method to be written to the Microlab STAR deck layout TubesToPlatePipette lay Instruments x o Instrument Short Instr Hame Deck Layout File Low Level si W Microlab STAR ML_STAR TubesToPlatePipettel 9 f Hidden Click on Browse to select the deck layout TubesToPlatePipette You can easily write the method by dragging icons from the toolbox on the left and dropping them in the method window on the right Finally your method should be as displayed in the next screen Method Load from ML_STAR Smart Steps Instrument short name ML STAR load s sequence ss ML_ST AR Tips ML_STAR Samples ML_STAR Plate ry Pipette with Tips from ML STAR Smart Steps Bk Instrument short name ML_sTAR standard pipette procedure Mode simple 1 1 Pipette volume HI 50 5 as sequence ML_STAR Samples controlling sequence Dispense sequence WML_STAR Plate Unload from ML_STAR Smart Steps Instrument short name ML_STAR unload s sequencers MIL STS Plate ML_STARSamples ML_STAR Tips The method TubesToPlatePipetie The deck is now loaded using the SMART Step load Sequence si Hide details Sequence Read tip Star pos 1 I No of pos 1 Reduoible
96. ation Now we enter the commands to load carriers onto the deck If your instrument is equipped with the Autoload option the carriers are automatically moved from the autoload tray into particular positions on the deck If not you will be requested to load the instrument manually In Keeping with the deck layout we will load three carriers the first holding the source plate the second holding the target plate and the third holding the tip rack 610766 02 Page 205 of 225 Microlab STAR User Manual Under ml_ star drag LoadCarrier to a line below ml_star nitialize A dialog window appears For the first carrier enter carrier position 6 and select rack type PLT CAR _L5AC Click OK This selects a carrier CAR for 5 deep well or archive AC plates PLT in landscape L orientation Again double click or drag LoadCarrier to a line below the last Load Carrier command A dialog window appears For the second carrier enter rack position 12 these carriers are 6T wide and select rack type PLT CAR_L5MD Click OK This selects a carrier for 96 well or 384 well microtiter plates Again double click or drag LoadCarrier to a line below the last Load Carrier command A dialog window appears For the third carrier enter rack position 18 and select rack type TIP_CAR_480 which is a tip carrier Click OK In the method frame we see ml_star LoadCarrier Position 6
97. bers read are assigned to their corresponding variables automatically Now define the file structure One line in the column specification of the file opening dialog represents one column in your file Click on the Add button to add the next line to the dialog Enter the data as given in the foregoing screen A header a variable and a variable type are assigned to each column The maximum column width is 20 for the string variables which are of interest here 610766 02 Page 149 of 225 Microlab STAR User Manual Select the variable sqlSelect from the dropdown list as a command string Click OK to finish the definition A loop is started looping over the input file until the file has been read completely E Opened Files Within the loop the first record of the file is read Fg File Read Edit File handle Cancel Help Next using the library function SeqAdd a sequence position is added to the hit sequence No if then is needed here because the SQL selection string automatically skips all records with absorption values less than or equal to the value of the threshold variable W SeqgAdd of HSLSeqLib Edit Library name C Programme HAMILTON Librar HSLS egLib hal Vanable Function name Fega P Function parameters Parameter Description O OS SeQUeENcCeOb ML_STAR Hits abwarel postion Then an entry is made into the methods trace file Hit
98. bid LabID String 20 posid Poslb String 20 od OD Float Command string sqleelect over following Tiles filet jooplounter2 used as loop counter variable File Read Read fron file tilet Seg dd of HSLSeqLib SeqAdd ML_STAR Hits labid poosid TreTraces of HSLTrcLib TreTracest Found hit labid labid poslIb poosid og og 9 End Loop 10 11 Load fromML STAR Smart Steps Instrument short name ML_STAR load s sequencels ML_ STAR Tips ML_STAR Hits ML_STAR Target Pipette with Tips from ML_STAR Smart Steps Instrument short name WiL_STAR Standard pipette procedure Mode Simple 1 1 Pipette volume pl 30 Aspirate sequence WL STAR Hits controlling sequence Dispense sequence MWL_STAR Target 12 13 q4 Unload from ML_STAR Smart Steps Instrument short name ML_STAR unload F sequencers ML_S5TAR Target ML_STAR His ML_STAR Tips Method Editor HitPick med Page 146 of 225 610766 02 Microlab STAR User Manual The first step in this method is to request the user to input a threshold value for the absorption threshold This threshold although numeric is entered as a string This is because in the next step a valid SQL database selection statement is created a User Input Edit Dialog title Weer Input Retur value Sou
99. ble tips Teaching mode refers to the possibility of teaching positions on the instrument deck which can be done moving the arm by the 610766 02 Page 47 of 225 Microlab STAR User Manual keyboard or by a teaching box accessory The channel to be used for teaching is to be specified too The next tab refers to barcode handling Error settings Step selection Instrument configuration Barcode type ISBT Standard w Code 128 Subset E and C v Lode 39 Codabar v Code 2 of 5 Interleaved 7 UPC AE 7 JAN EAN 5 Trace generation W Barcode trace Sample tracking The different barcodes which are allowed on the instrument can be selected here In addition the generation of a barcode trace file barcodes_N txt can be switched on and off storing rack barcode and track position on deck as well as tube or plate barcodes and positions within the rack The sample tracker which generates a database of liquid transfers in the background can be switched on here too This sample tracker is used to generate a Microlab AT like barcode file using a special filter tool The error settings tab allows you to specify the step width in mm for blood clot error recovery the channels move up on request by the step width The return string separators are used to define the syntax of single step return values Error settings Step selection Error settings Blood clot step width Return string word separator Retu
100. ce will be re ordered by selecting the i th grid row and moving that row to the next position H Group ever TOWE Start at row Sort alfa eel Sort Racks A Sortby Grid Columnes 4 Sort by Grid Rows Filter by Probe Head 7 1 6 Filter by Probe Head The Filter by Probe Head dialog allows you to a apply an order to labware positions in the active sequence then b remove positions where the probe won t fit These functions are applied to the currently selected labware positions or to the entire sequence if no positions are selected The first function a is optional Initial values for this tab are taken from the instrument s configuration file These initial values can be recalled using the Defaults button Els Select traverse direction Number of rows of tips fi Number of columns of tips fiz gt a i Distance between each row fo mm Distance between each column E mm gt 2o Tipi row number fi Tip column number fil gt Serpentine Filter Defaults lf ad f Sort Racks A Sortby Grid Columns A Sort hy Grid Rows A Filter by Probe Head Page 62 of 225 610766 02 Microlab STAR User Manual 7 1 7 Sorting Racks Example Here is an example of how rack sorting works Given the following sequence across 2 racks apply the Sort Racks filter defining a direction across the racks aj met lay Hamilton Sequence Editor File Edit View Sequence Play Help Eie Edt Vie
101. ch sample Volumes Pipette volume pl Tip counter 100 Tip counter T Write tip counter Pre aliquot volume pl Rest volume pl fa 0 Redizpense height from bottom mm 9 f Disp to waste fa Disp to aspirate sequence Channel settings Error settings f Aspirate sequence is controlling Aspirate sequence MLS TAR tubes T Reload i Reducible by user f Not reload Dispense sequence is controlling Dispense sequence MLS TAR MyPlate a Reload iY Reducible by user f Multiple use Dispense mode EENES Advanced i Jet Surface I Aspirate all i Advanced Liquid liquid class oMso High allume DMSO Dispense et T Finish Cancel Help Let us now walk through the different sections of the SMART Step 610766 02 Page 71 of 225 Microlab STAR User Manual General Instrument short name ML_STAR Standard pipette procedure Aliquot procedure Pipette mode f Simple 1 1 Replica 1 n Multiplicity r Pooling r 1 Standard or aliquoting mode must be selected For standard pipetting mode three different procedures are possible Simple pipetting for simple aspiration dispense cycles Replica for cycles of aspirations dispenses where the liquid from one source is dispensed into multiple target containers no aliquoting e g for a double test tube 1 gt plate position A1 A2 tube 2 gt plate
102. ck 4 6 NOTE There are only two wash stations allowed on a deck Each wash station must have its unique identifying number Each washer over both carriers should have a unique identifying number At edit time in the wash step the user can only select sequences out of the list of sequences with underlying racks having these properties 610766 02 Page 201 of 225 Microlab STAR User Manual 17 5 Z Positions of Carriers Racks and Containers The aspiration and dispense dialogs always use the inner container bottom as a reference position fixed height liquid level 0 The x y z values of the reference well usually A1 or 1 labelled red in the move labware dialog stored in the instrument s system of coordinates are shown in the Move Labware dialog To access this dialog right click the labware item of interest in the Deck Layout Editor and select Move Labware The rack and the container both have a clearance height which means that the movement of the channels is not impeded if they pass above this height The software automatically takes the highest clearance height The maximum pipetting height is counted from the bottom of the container upwards and determines the dead volume of the container The LLD search height is the height at which the speed of the channel is reduced to look for the liquid surface Clearance heights of rack and container LLD search height A Maximum pipetting height the channel c
103. d on to the instrument deck Click OK If you want to decide at runtime which or how many of the wells of the source plate are to be transfered to the target plate click on Show Details and enable the checkbox Reducible for the source sequence Hide details Sequence Reed Start pos Wo of pos 1 Reducible Click OK again For an instrument with autoload option this command loads the carriers automatically onto the instrument deck during runtime For a manual load instrument this command requests the user to load the carriers for runtime Page 120 of 225 610766 02 Microlab STAR User Manual Now program your pipetting Drag the SMART Step pipette to the line below the loading step MH Pipette with Tips Edit General Tip sequence Instrument short name 5 ML_STAR Tips Standard pipette procedure Aliquot procedure IY Reducible by user Pipette mode Simple 1 1 C Replica 1 n C Pooling n 1 Tip handling replacement Multiplicity n After each dispense One tip set E After banstered volume None After each sample Volumes Pipette volume pl Tip counter a0 Pre aliquot volume pl Rest volume pl fa T 0 Tip counter Write tip counter Redizpense height from bottom mm Disp bo waste fa Disp to aspirate sequence Channel settings Error settings f Aspirate sequence is controlling Aspirate sequence ML
104. der advanced pre rinsing and liquid following can be specified Dispense Advanced Fa Frerinsing Mis settings Initial sequence manipulation Set current position to first sequence position Set end position to last sequence position Final sequence manipulation f Sequence conesponds to range remaining after this step Sequence conesponds to range used within this step Enable prennetag mir Cycles Mis position mm Yolume pl x Miscellaneous calibrate camer if supported Cancel Help Furthermore sequence manipulations can be made separately for aspiration and dispense sequences If the sequences which this SMART Step is going to work with have been used within this method in preceding steps the sequence counters can be reset using the two check boxes for initial sequence manipulation If other steps are following this SMART Step the status of the sequence which is passed back can be defined by the radion buttons under final sequence manipulation Under channel settings the user may specify a channel pattern manually as well as the number of the channel which will be used to calibrate 1536 well carriers The error settings dialog allows you to specify an error handling approach Error Settings Error handling C Abort method T Copy pattern T Exclude error positions Walk away mode Timeout 2 T Enable Cancel Help The choices in case of an
105. dinate System 4or8 775 775 465 12 nas 1135 393 16 149 5 1149 5 321 The random access range of the different numbers of channels is indicated by brackets at the left side of the deck layout display in the User Software see chapter 6 for an account of deck layouts Page 24 of 225 610766 02 Microlab STAR User Manual To guarantee random access to sample carriers only the inner tube positions should be used The number of blank positions to be used for the different instrument configurations is listed in the following table No of ___SMIP CAR 32_ SMP CAR 24 SMP CAR 12 Front Rear Front Rear Front Rear Channels Blanks Blanks Blanks Blanks Blanks Blanks aos o o o o o o a 3 3 2 2 4 16 5 5 4 4 2 2 The 16 channel Microlab STAR is intended as a batch type processor i e random access to all positions is not possible By contrast with an 8 channel Microlab STAR the methods for a 16 channel instrument should be set up in such a way that all positions are processed in batches of at least 8 or better 16 to optimize the pipetting speed with 8 or 16 simultaneous aspirations or dispenses at identical x coordinates see section 8 9 for more NOTE If pipetting positions outside the random access range of the instrument are used the system may crash However a strictly batch type process can eliminate these problems 3 2 2 High and Low Volume Channels Different pipetting heads ar
106. dit to open the Liquid Details window and go to the Liquid Details tab Edit Liquid Class Standard olume_Water_DispenseJet 1 0 Liquid water Tip type Standard volume tip without filter Dispense mode Dispense jet Liquid parameters 300 pl 1000 pl Asoirat Dy RRS RENSE Channel Channel Flow rate 100 180 0 4 250 1 500 ia 0 4 250 1 500 LT Mis flow rate ir transport valume 50 50 0 50 0 50 0 3 160 0 3 160 DI mm TTT Blowout walume Swap speed Settling time 0 2 2 Over aspirate volume 18 Clot retract height 0 50 Stop flow rate 0 4 250 Stop back volume 18 Set Version i Original Liquid Class Abbrechen Hite Liquid Details Window The Liquid Details tab has two sections At the top the attributes of the liquid class are shown Liquid Name tip type dispense mode NOTE A liquid class is valid only for the defined set of attributes The liquid parameters section on the left specifies the appropriate instrument parameters for aspirating and dispensing Here is what the various parameters mean e Flow rates and Mix flow rates are volume flows of liquid in ul s they correspond to plunger speeds for aspirating dispensing and mixing e Air transport volume air for transport is aspirated at the end of the aspirate step and automatically dispensed again as an extra v
107. dow Help x PLACE LEELLE RE device ml_star HS5L_OneFlateToFlate lay Do not insert text before this line Calibrate 1536Plate aati TransferPlate Dispense ml star Initializef 3 FirmwareCommand tl star LoadCarrierf 3 Initialize ml star LoadCarrierf J oadCarrier ml ostar LoadCarrier jj Move loPosition TipEject TipPickUp ml_star TipPickUp 3 UnloadCarrier nl_star Aspirate tl star Dispensef j tl star TipEject 1 til star Source il star Targett ml star Tiptt while i ml_star Source GetCurrentPositionf 0 3 return library functions an x Hja Syntax Chech Now save your work by selecting Save from the File menu The Software prompts you to perform a syntax check Click YES The result of the syntax check is displayed in the bottom frame of the method editor If you have typed in the commands correctly the result should be O error s Another example is available within your demo methods directory copy from tubes to plates 610766 02 Page 209 of 225 Microlab STAR User Manual Appendices A Glossary Adjustment Detailed positional setting for the hardware Air displacement tip Commercial pipetting tip Aliquot Aliquots are identical small volumes of liquid Aspirate To draw up liquid into a pipetting device Autoload Option and hardware assembly that enables automatic loading of the Microlab STAR It consist
108. e 84 of 225 610766 02 Microlab STAR User Manual NOTE To change between the different views right click within the tool window and Select tree view The table on the next two pages gives a brief outline of the available ML STAR specific commands These instrument specific single commands are used in both the Graphical and the HSL Method Editors 610766 02 Page 85 of 225 Microlab STAR User Manual Command Icon Action performed Parameters to be specified Initialize Initializes the instrument Waste destination channel Load Carrier Loads a carrier on the deck Carrier position rack type Unload Carrier Removes a carrier from the Carrier position deck Tip Pickup Picks up a CO RE tip or needle Sequence tip type channel Tip Eject Discards the tip into the tip Waste destination waste or the needle into the channel wash station or rack Aspirate Draws liquid from a container Sequence volume liquid name channel mix settings LLD The carrier position gives the track number on the Microlab STAR deck where the carrier will be inserted For microtiter plate carriers which are 6 tracks wide the carrier position refers to the leftmost edge of the carrier Page 86 of 225 610766 02 Microlab STAR User Manual Command Icon Action performed Parameters to be specified Dispense Drops liquid into a container Sequence volume liquid name channel mix settings LLD Calibrate 1536 Measures precise pos
109. e aspiration the check box aspirate all enables aspiration of the residual liquid from a container specify a volume larger than the expected residual volume within the container On dispense the dispense mode surface or jet has to be selected For aspiration and dispense the LLD liquid level detection settings have to be specified by clicking on the LLD button Aspirate Liquid Level Detection LLD Capacitive LLD Pressure LLO T Enable pressure LLD Sersitrvity f fern high i fiddle D Aigh CD iay f Yer high C Frade D Aigh EE iay Detail settings Submerge depth mm Aspirate height from Gotkom mm Mas height difference mm a es Cancel Help Within the LLD dialog capacitance pressure or both LLD types may be switched on A sensitivity setting is necessary see table in chapter 2 If LLD is used a submerge depth has to be specified if a fixed height is prefered the distance counted from container bottom is to be specified For the parallel use of both LLD types a maximum height difference has to be given within which both LLDs have to respond The next step is to select the appropriate liquid from the dropdown field Liguid liquid class DMSO High olume_DMSO_Dispenselet The selection by attributes tip type dispense mode is automatically made so that only the liquid name has to be selected 610766 02 Page 77 of 225 Microlab STAR User Manual Un
110. e available for the different volume ranges a from less than 1 ul to 300 ul for the use of low volume tips 10 ul and standard tips 300 ul or b from 5 ul to 1000 ul for the use of standard tips 300 ul and high volume tips 1000 ul Currently a mixing of high and low volume channels on one and the same instrument is not feasible 610766 02 Page 25 of 225 Microlab STAR User Manual 3 2 3 Autoload Option The Autoload option like the options mentioned above belongs to the Microlab STAR configuration initially specified by the customer and cannot be added later Barcode Reader The Autoload option and barcode orientations for tubes and plates The autoload unit is a device enabling automatic loading of the Microlab STAR It contains a loading head that e moves in an X direction e shunts carriers onto and off the Microlab STAR and e reads the barcodes of carriers tubes and microtiter plates There is a presence sensor that identifies the tubes present on a carrier Barcode reading is only possible in conjunction with the Autoload option Equipped with autoload the ML STAR can read the following barcode types ISBT Standard Code 128 Subset B and C Code 39 Codabar Code 2 of 5 interleaved UPC A E JAN EAN8 Page 26 of 225 610766 02 Microlab STAR User Manual NOTE In addition barcodes must have a minimum readability i e good contrast size correct orientation and distance between bars
111. e carriers with the left mouse button You now see the 96 selected wells of the four carriers The grid window on the left shows the generated sequence of 96 positions holding all tubes Jtebware i car24 cuplss10 car24_cuptsxt0 7 0 2 3 4 oar24_cuptsxt0 1 Position ID TOO 0D DT Dd D d d d oOo oOo aie Em Enim Ea Cee e e m peie Em Enim Ea Ce a a a pnie Em Enim Ea Ce ee a eie Em Ee Em Ea En e a e e ce Seeded Ce a a a FATE ii PRR TT ETE i E oOoodooOoooooo eee ee eee ae ae ae aa a Mi iear24_cupisxt021 22 car24 cuplss10 22 K a 15 20 29 Select Save Sequence from the Sequence menu and enter the name Samples for your new sequence Select Save from the File menu Now the sequence linking all samples is stored with the deck layout Select New from the sequence menu Now create a sequence linking all tip racks by rubber banding the tip racks Save the sequence under the name Tips Now create a corresponding sequence for the plate again by rubber banding Save this sequence under the name Plate Select Exit from the File menu then click Yes in the pop up window to save the changes to the deck layout Technically sequences are part of the deck layout Select Save from the File menu within the deck layout editor to save the changes to the deck layout under the name TubesToPl
112. e count 3 Comment Create a Microlab 47 Barcode Format for plate Create TBarcodetile of H gt LML_STARLib HSLML_STAR Create 1Barcodetile ML STAR plate The sample method Firstly initialize the system and load all carriers using the single steps Load Carrier New Carrier to load Advanced Templarmner Error settings coed Hee 610766 02 i Channel 1 0 11111111 Optimized channel use All sequence position Sequence ML_STAR AlTips Sequence counting 1 Automatic Channel 1 08 11111111 Optimized channel use All sequence position sequence ML_STAR AllTubes Sequence counting 1 Automatic Liquid name StandardYolume_VYater_DispenseJet Volume 220 ul Mix volume pl Cycles 0 Position relative to liquid surface mm LLO Channel 1 5 11111111 Optimized channel use All sequence position Sequence ML_STAR plate Sequence counting 1 Automatic Liquid name Use the same liquid as aspirated Volume 220 ul Mix volume pl Cycles 0 Position relative to liquid surface mm LLD Channel 1 08 11111111 Optimized channel use All sequence position Sequence ML_STAR Vyaste16 Sequence counting 0 Manually Dialog Title Output Return value OutputReturn Buttons Yes and No button Default Mo Icons Display information message icon Page 159 of 225 Microlab STAR User Manual Assign the name of the tip counter MyStandardTips to be used t
113. e driven by methods so it is important that they are correctly defined and easy to operate Programmed methods are stored as a linked set of ASCII files in the Methods sub directory on the hard disk They can be opened edited and saved either in the text like HSL Method Editor 9 HSL Method Editor page 96 or in the Graphical Method Editor 8 Graphical Method Editor page 66 The present chapter takes a cookbook approach It teaches you how to program a number of simple methods typically used in laboratories By following these steps you will become familiar with the layout and workings of the software and can then modify the suggested methods to suit your own particular requirements or program new methods based loosely on those suggested The scope of the complete command language is rather like that of common higher programming languages For details refer to the Microlab STAR Reference Manual as incorporated in the online help NOTE Sequence definitions have an important influence on programming methods AN ATTENTION Ensure that all methods are tested first using deionized water ATTENTION The user is responsible for the validation of the method All sample methods explained in this manual are available in the DemoMethods_MLSTAR directory following installation of the Microlab STAR User Software 13 2 Create a Method to Copy from Plate to Plate Using SMART Steps The method we are going to describe first copies
114. e empty container so close as to allow the dispensed droplet to have simultaneous contact with the tip or needle and the container bottom Trace Note of a status during a processing Tube A narrow container for liquid usually having a circular cross section and a cylindrical length section User User of the product Levels of user proficiency are not distinguished by the system Verification Kit Option Aid to check the functions of the Microlab STAR including adjustment Page 214 of 225 610766 02 Microlab STAR User Manual Waste Container A device on the Microlab STAR deck to collect used disposable tips Well The individual hollow in the MTP Well type Geometrical shape of the well such as U V or flat Wick side of container Type of dispensing whereby tip or needle touches side of container and thus releases the droplet Not possible with Microlab STAR Work area The area on the Microlab STAR to which access is provided during processing Elements to be pipetted can be placed in this area Worklist Information sent from outside the system as to what method s is are to be executed on the Microlab STAR and with what liquid 610766 02 Page 215 of 225 Microlab STAR User Manual B Index Access rights 4 52 Accessories 1 3 29 32 Adding labware 4 59 60 61 62 120 184 206 216 Air displacement 3 14 25 Aliquot 6 23 25 28 80 82 83 84 87 155 157 158 185 23
115. e following carriers car standardtip 0001 cart4 cup 5s100_0001 cared cupl5e100_0002 cared cupl 5x1 00_0003 card cup 5x1 00_O0004 plt_carlBrmd_O001 nun SE fl_ SR RRR RPE TETIT ITIITI TITTI s O sene O me Leet Remove Ai Removed Remaining foe oo E a oo E E e a e E a a e o y e e o r G Heset Remove All je je Help One may know to reduce the number of samples to 24 The method then copies 24 tubes from the first carrier in this case to the plate and stops It is also possible to deselect distinct tubes from the sequence by clicking on the wells A reset button is available to restore the Original sequence An example of how to program this method using single steps is available on the user software CD 610766 02 Page 139 of 225 Microlab STAR User Manual 13 5 A Method to Pipette Aliquots Using SMART Steps In this example a simple aliquoting procedure is decribed The method aspirates 280 ul from a sample tube rack and dispenses the pre aliquot as well as 12 aliquots of 20 ul each into an empty plate The last aliquot is ejected with the tip Here s the deck layout nun SG fl_ Fi Oleg Place the following labware components on the deck a tip carrier TIP_CAR_480 with one tip rack for standard volume tips a sample tube rack of 24 tubes called Sample and standard plate carrier PLT CAR _L5MD with a 96 well flat bottom microplate e g the Nunc
116. e rear see picture 1 below The fuses for the instrument are situated in the mains power switch see picture 2 below Plug the mains cables for the computer and the instrument into the same electrical outlet Connect them only to an earthed outlet Voltage selector Always at 220 240 V ATTENTION Always keep the instrument voltage selector at 220 240 V Place the appropriate fuse in the mains power switch before switching on the instrument The technical specifications regarding electrical power are listed in the following table Maximum power consumption 600W Voltage 115 230 V 15 15 Frequency 50 60 Hz 5 Delayed action fuse 115 V 6 3 A 230 V 3 15 A Overvoltage category II 610766 02 Page 37 of 225 Microlab STAR User Manual 3 6 2 Needle Wash Station The needle wash station has its own power supply The mains plug is on the rear of the pump station see picture below Ensure that the the needle wash station is correctly earthed when connected to the power supply Connect the wash station only to an earthed outlet Ensure that the voltage selector is correctly set before operating the needle wash station The fuses for the instrument are situated next to the mains power switch see picture below The pump station has two fuses for the power supply which can be accessed by opening the cover above the mains plug Voltage Selector Fuses for power supply Fuses for power supply o
117. een sites on one carrier by simple drag and drop Besides plate and tip carriers tube carriers can also be placed on the deck These carriers are preloaded with tubes of a defined size diameter and length In fact these carriers are racks rck in contrast to the other carriers which are carrier templates tml The tube carriers are an example of racks which fit directly into the track grid of the Microlab STAR Page 54 of 225 610766 02 Microlab STAR User Manual 6 2 2 Adding Labware Directly to the Deck To place labware e g microplates directly onto the deck without using a carrier select Add Labware from the Edit Menu A dialog window opens up Select the type of labware from the library and enter the x y z coordinates by hand You may enter a name by which the rack will be Known on the deck In the type field select the already listed labware item or press Browse to load another laoware item to the list Click OK If the labware position falls outside the instrument deck a message window opens to inform you Add Labware 21 x Name MyNewLabware Type Browse Orientation Lab Editor Rotate Fixed deck Position 5 C Free form coord mm Y coord mm Z coord mm First position fo fo fo Move Probe Position probe at center and bottom of container at first position m Deck position Last position fo f f P
118. elect the sequence ML_STAR WashStations Drag a loop from the general commands f berate over sequences e e S O ML_STAR Plate E MLSTAR Samples TT ML STAR Wiashstations L J me n mai Loop over the Samples sequence Page 154 of 225 610766 02 Microlab STAR User Manual The next step is the adjust sequences command This command is used to automatically handle a reduction of samples when using single steps If say only 4 samples are left over in the last step adjust Sequences ensures that the system will pick up only 4 tips aspirate 4 samples and dispenses into 4 plate positions leaving the other 4 channels unused zh Adjust Sequences Edit Sequence s Bip ee WL STAR Plate ree wo 3 ML_STAR Washstations iS t lt CS F Add Remove Cancel Help Multiple sequences may be added to the dialog by clicking on the Add button and selecting the sequence from the dropdown field NOTE Every sequence that is used within the loop has to be added to the adjust sequences command including the tip sequence The position of the adjust sequences command within the loop line no is not important as long as it is placed within the loop Now pick up needles using the SMART Step EM Needle Pick Up Edit Instrument short name Needle sequence ML_5 TAR Wash Stations Channel settings Error settings Cancel Help 6107
119. equence counting lid sequence Transport mode plate sequence sequence counting lid sequence Initialization must be the first instrument specific step in each method Page 88 of 225 610766 02 Microlab STAR User Manual 8 3 Writing a Method You can now easily write a method by dragging icons from the toolbox on the left and dropping them into the method window on the right f3 Hamilton Method Editor MyDemo Oy x ED Fle Edt view Method Tools window Help 0 s Sex mee gia e s Foe boop KF over following Sequences ML_STA nunc_S6_flat_l Mo loop counter variable used TipPickUp on ML_STAR Channel 1 83 11111111 Optimized channel use on Sequence ML_STAR hamilton_96_standardyvolumetip_i Sequence counting 1 Automatic Tip type Standard olumeTip Return value count 1 Aspirate on ML_ST A amp R Channel 1 11111111 Optimized channel use on Sequence ML_STAR carrier wih _24_cup1 5x951 Sequence counting 0 Manually Liquid name Standard y olume Vater _DispenseJet Volume 50 pl LLO settings off Liquid following an Return value count 1 Dispense on ML_STAR Channel 1 8 11111111 Optimized channel use on Sequence ML_STAR nunc_96_flat_l Sequence counting 1 Automatic Liquid name Standard yolume_VYvater_ DispenseJet Yolume 50 pl LLO settings aff Liquid follovying on Return value court 1 TipEject on ML_STAR Channel 1 11111111 Optimized
120. erminates a loop If then else Condition command Commands on line 3 Sequence Get current position Sequence Set current position Sequence Get end position Commands on line 4 Sequence Set end position Sequence Adjust sequences Open file To open a file before file operation Commands on line 5 Read from file Read data out of a file Write to file Write data into a file Set position Set the file pointer to a specific position Commands on line 6 Close file To close a file after file operation Timer start a timer Timer wait for a timer Commands on line 7 Timer read elapsed time from a started timer Timer restart a timer User Input display an input box at a particular point in the program Commands on line 8 User output display an output box at a particular point in the program Communication Port Open Opens a com port Communication Read Reads data from a com port Commands on line 9 Communication Write Writes data to a com port Communication Port Close Closes a com port Shell call external program Page 68 of 225 610766 02 Microlab STAR User Manual Commands on line 10 Wait for a event waits for an event Set event defines an event Abort Abort Method Commands on line 11 Error handling by user Defines a section with user defined error handling Begin Parallel Starts a bifurcation for programming parallel processes End Parallel Ends
121. error are abort the method only abort button on runtime error dialog give the choice in runtime to abort or to cancel continue with user defined error handling if programmed otherwise abort or use the fixed default error recovery recommended pre programmed for the SMART Step In this case two choices can be made Page 78 of 225 610766 02 Microlab STAR User Manual Copy pattern means that in case of an error on aspiration the corresponding well of the dispense sequence will be left out the pattern is kept If copy pattern is not checked the dispense sequence positions will all be pipetted leaving an unpipetted well at the end of the sequence Copy Pattern Checked Copy Pattern NOT Checked Exclude error positions means exclude the erroneous positions from aspiration and dispense sequences If this option is enabled and an error occurs during an aspirate or dispense step the erroneous position will be excluded from the sequence by removing the corresponding element from the aspirate or dispense sequence The next time these sequences are used the erroneous well will not be pipetted In addition walk away mode can be enabled If this checkbox is checked a timeout has to be specified after which the error dialog on runtime will automatically close down again to continue with the selected error handling Abort Cancel or default error recovery NOTE Exception If the option default error recov
122. ery is chosen and an error occurs for which the default is set to abort e g a hardware error the walk away mode is left on and the dialog waits for user interaction The other default settings correspond to those of the single steps see chapter 15 4 for further explanations of walk away error handling 610766 02 Page 79 of 225 Microlab STAR User Manual 8 2 1 2 SMART Step Load Unload This SMART Step is used to automatically load the instrument deck with all labware necessary for the run These commands work with autoload and manual load instruments For a manual load instrument the user is requested to load unload the carriers by hand Dragging the load command to the method tree opens up a dialog box Mi Load New Instrument short name ML_STAR Sequencels Show details Sequence OO Readtip counter Tip counter 1_ ML_STAR MyPlate z ML_STAR tips_st 3 MyStandardTips Add all sequences Add Remove Carrier calibration Error settings Channel number W Calibrate if supported a Cancel Help The procedure for loading the instrument follows logically from our concept of sequences Whatever carriers have to be loaded to ensure that the sequence which is used within the method is available will be loaded If for example a sequence of samples spans 3 tube carriers all 3 carriers will be loaded automatically to their locations tracks as defined in the deck layout The d
123. ew carrier within the labware editor Now open the deck editor and place the carrier onto the deck nun _96_ fl nun _96_ fl fl w 10 15 ral ra The carrier is now preloaded with a plate on its site Page 196 of 225 610766 02 Microlab STAR User Manual Furthermore a carrier has several properties such as barcode positions which may be specified To do so open your newly defined carrier in the labware editor and select Edit gt Properties A dialog opens up Here we open a standard plate carrier Labware Specific Properties Property Ital Value MlStarlarBCOrentation MlStarlarBlReadvyidth 300 MlStarCarCountOtBCPos 45 MlStarlarFirste Pos MlStarCarls4utoLoad Modify M StarlarlsLoadable MiStarCalsRecagnizable 1 Add MlStarlarPos4reHecog 0 MiStarCarh asterw idth J60 Delete MlStarCarwidth sT B Cancel Properties of a PLT_CAR_L5MD Entries can be added by clicking on Add The entry names are to be kept they roughly explain the property see chapter 17 4 Add the entries and click OK to store them along with the carrier NOTE Always start with the given settings of a standard carrier and apply changes step by step Do not change the names or typing of the variable names These variables are system variables 610766 02 Page 197 of 225 Microlab STAR User Manual 17 4 System Flags for Labware Properties Within the labware some flags and settings labware properties a
124. f 225 610766 02 Microlab STAR User Manual MWe FOE Oy GERON cece ye la das daca eace tent deacanse a cearihe fusednen vacounat eacanseatcnanhe se anda onaaastduaaaeteneanhesnauesset 62 LEO Piter Dy Prope Cad saer uses onanciiatneecshatatintoe secs seanuraanuanceatiueec ented staunalseheandinesoeans 62 TEE SOME TRACKS KAM Gasca chasis ead tnrinrst en saint nace tea ea cuicsrduaihe Bwana E Oaeisaaneeuh 63 TAGS Sorte Grid Rows Example iicg iG es anata S a ean acnaninsumuiees 65 Graphical Method E CUOF csie EEE EN 66 SL General Method Commands eisiaa naaa eaa o EAE E Eae 67 8 2 Using Microlab STAR Specific Steps scssssssseecccocsssssccccocsssseecccoossssseeccocssssssccecssssseseeesssssoe 69 Sll SMARTS D uoaa e SrA Pe eR eee 70 B22 oE O SOPS aa a a stance loa osu seach lanl ata cearme aenaseulauaeeatees 84 S3 WMS a Method sicssescceesswecdectassuseeustancvsssscndeussanvscieebasevssseusussweanvosussesiscususeudessdecdvcarssasecustescees 89 95 4 Editine a Metho issiir eona a E O 89 Sed V ANIA DIOS E E E EAE E E E E EN E 90 8 6 Using Library Functions the ML STAR Library sessccsssssseeccocsssssceccoosssssscecosossssscecessssoo 91 8 7 Binding Instrument Step Return Values cccsccccccssssssssscccccccccssssssscccccsscesssssscccooes 93 9 9 Workin with W OrFKIISts wi decsscciensiiaiacassiccevesenstacssaseesussenasessesucedivvseseesassuadanvoansessacveaadecassusdseeve 93 Soal Pile PORnMats yscecsaued tastes E anced
125. f their characteristics are specifically intended by their manufacturer to be used for in vitro diagnostic examinations 610766 02 Page 219 of 225 Microlab STAR User Manual Declaration of Conformity We Hamilton Bonaduz AG CH 7402 Bonaduz Switzerland confirm that the following product Product name S N meets the following EC directives including all applicable amendments Applied harmonised standards Safety Emission Immunity Additional Information Ti US C Page 220 of 225 Wir Hamilton Bonaduz AG La soci t Hamilton Bonaduz CH 7402 Bonaduz Schweiz AG CH 7402 best tigen dass das Bonaduz Suisse confirme folgende Produkt que l instrument ci dessous Microlab STAR mit den folgenden EG est en conformit avec les Richtlinien einschliesslich directives CE suivantes y aller zutreffenden compris leurs Aenderungen amendements le cas bereinstimmt ch ant EMC Directive 89 336 EEC Low Voltage Directive 73 23 EEC EN 61010 1 1993 A2 1995 EN 60825 1 1994 A11 1996 EN 61326 1997 A1 1998 class B EN 61326 1997 A1 1998 laboratory equipment Canada USA CAN CSA C22 2 No 1010 1 92 UL Std No 3101 1 FCC Part 15 class A 610766 02 Microlab STAR User Manual D Ordering Information A current checklist is available from HAMILTON 610766 02 Page 221 of 225 Microlab STAR User Manual E Principles of Calibration Statistical De
126. f wash station The technical specifications regarding electrical power for the wash station are listed in the following table Maximum power consumption 140 W Voltage 115 230 V 15 15 Frequency 50 60 Hz 5 Delayed action fuse 115 V 3 15 A T3 15L250 230 V 1 6 A T1 6L250 Overvoltage category II Page 38 of 225 610766 02 Microlab STAR User Manual 3 Maintenance A short preventive maintenance including volume verification should be carried out twice yearly Typically the service and volume verification are carried out by a trained HAMILTON service engineer To clean the instrument deck use the procedures required by GLP To preserve plastic materials from damage do not use organic solvents If a spray is used do not point the spray directly at the autoload unit or at electrical boards or connectors 3 8 Verification The Microlab STAR will be verified by a trained HAMILTON technician upon initial set up and thereafter at regular intervals for a period of time specified by service agreements HAMILTON recommends that this verification take place four times yearly For volume verification in the field HAMILTON will supply a verification tool based on gravimetrical measurements the gold standard of volume verification The detailed specifications are listed in the specification table at the end of this chapter Conditions are valid only for the HAMILTON verification kit 3 9 Disposal Af
127. fesaiend A E ideeectenateactanseeestueiieacs 93 8 0 2 Worklist Handling with Microsoft Excel ariasi ini ctierien ese eae as 93 8 9 Writing Batch type Methods for a 16 Channel Microlab STAR ccccssssssssssssceeoees 95 w FIST MCN Od EANO AEAEE E ET 96 LO Ruw CONT OL osis E E E 98 ROSE Meth d Linking oiesssciacesciecoecsesedarsastwsecsvesadavekstwancevssalosvepbusocsesiv sasusunanssseiviaseeniwsodsseswaguettenes 100 11 The Microlab STAR Lig id E dilok sisenes E E E E EA 101 ELD Conceptof Liquid CASS eaiicsccacccsccossscssens ccccwasevscsvasssccesiovsecsiosasooussedencs daasectsNocssesiaseccesses 101 12 Editing Liquid Detas wiiiccssctesasvessswstatusuces ses cisslecstacastacccucduuasavesssuciseloastesdstesasweseteiwissdsiaoees 101 11 3 Defining a Custom Liquid Class siisscsscseccccssssnssassdeccesscasedessedecscsscvsncvosetwsdccesudecdsseioacseseabecs 104 11 4 Importing and Exporting Liquids ssseseeecoocsssseccccccsssssececosossssceccocsssesseceossssssesessosssoo 105 12 Microlab AVS Automated Vacuum System cccrrrrsscccccrrsssscccccccssssccccccccssssccccsccssscsceees 106 I2 INOS ratlom Ol Ihe AVS aiaa a E a e aoa 106 122 Commands torth AV Sesera ear Oe Eea eaaa eana 111 12 3 Paralel Processine with the AVS cc disiiecciescdboscssensctiesccssedacdsevetoccecocsaseeusaleadecsedacesetadoodoeets 114 13 Methods for Microlab STAR visscccccccscssscsssssssssssssssssssssssssssssssssssscssssssssscsssssssssssssssssssssees 117 RA E o
128. finitions Used for Calibration Calculated value in mi x UL Xp_ mE x mE where x measured standard value Xm Median of measured cuvette X target value Mean x M Te il gt 1 II Standard Deviation s Accuracy A where f target value Page 222 of 225 610766 02 Microlab STAR User Manual F Needle Wash Station Chemical Compatibility 1 Chemical compatibility of standard wash station Acetic acid 20 1 2 3 i1 3 1 1 1 1 1 1 2 Aceticacid glacial 1 4 alilala lalil 41 4 4 e2 acetone lafa fa fo ae i fa fal i lola s acetonie 1 3 la i lal olals 1 lo 1 ol MO MUMMY aloes D 2 i i 0 D Chloroform 1 4 l a a al i lalas a1 fal a al Deionizedwater 1 1 a fafa i ait a fa fa al Dimethylformamide 1 2 4 1 4 1 1i4 1 4 4 2 Dimethylsuloxide 1 3 3 a 4 0 11 1 41 4 3 ol Ethylacetate 1 3 ajo 4 1 1ifa 1 4 3 2 Herne ot 4 ft of ilill a fa 1 al Hydrochloric acid 20 4 1 1 1 4 1 4i41 ililia a isopropyl acon 1 t a fa 3 a4 ait a fal a a metran 4 t 3 af 3 a a tf a fa a a Methylene chloride 1 4 3 4 4 2 Jalal ilal i la Nitric acid 510 1 2 1 fafa i1 4i1 a fafa e l Nitricacid 70 1 4 l2el lalal ilala ila i ia Phosphate buter 1 1 41 4 41 0 1 1 4 lolai a4 Phosphoric acid e5 2 3 1 1 4 0
129. following approaches e Use only HAMILTON tips on the Microlab STAR e Use new tips for every pipetting step to avoid carry over between different wells or containers e Use filter tips to avoid contamination of the pipetting channel by jets aerosols etc e Always dispense to a surface Dispensing in a jet may produce aerosols and thus cause cross contamination e Always dispense using a residual volume i e do not completely empty the tip on dispense This can be achieved e g by aspirating 10 ul and dispensing only 9 ul 2 4 Liquid Classes Pipetting Modes and More In general pipetting on the principle of air displacement as with hand pipettes is sensitive to e the manner of pipetting e g surface or jet e tip or needle type e environmental effects temperature pressure humidity and e liquid type The instrument s behavior is determined by specifying the pipetting mode e g surface or jet mode and the liquid class Pipetting mode and liquid class represent two independent sets of information which both have to be specified For aspiration three modes are possible e Simple Aspiration for all standard cases e Consecutive Aspiration for aspiration with a tip that has already aspirated liquid and e Aspirate All for aspiration of all the liquid within a cup specify a volume larger than what is expected within the cup In this case aspiration monitoring is deactivated and the tip will follow t
130. from ML_STAR Smart Steps Instrument short name ML_STAR Needle sequence ML_STAR VvashStations Consume sequence positions OFF Aspirate on ML_STAR Channel 1 5 11111111 Optimized channel use All sequence position Sequence ML_STAR Samples Sequence counting 1 Automatic Liquid name Standard olume_VYater_DispenseJet volume 50 ul Mix volume ul Cycles 0 Position relative to liquid surface mm LLD settings on Capacitive 3 Return value count 3 Dispense on ML_STAR Channel 1 033 11111111 Optimized channel use All sequence position Sequence ML_STAR Plate Sequence counting 1 Automatic Liquid name Use the same liquid as aspirated Volume 50 pl Mix volume pl Cycles 0 Position relative to liquid surface mm LLO settings off Liquid following on Return value count 3 Needle Eject trom ML_STAR Smart Steps Instrument short name ML_STAR Needle sequence ML_STAR VvashStations start wash ON End Loop ML_STAR Samples Reset Sequence atter loop Unload from ML_STAR Smart Steps OL Instrument short name ML STAR unload 2 sequence sy ML_ STAR Samples ML_STAR Plate a A method using needles Load the Plate and Samples sequences The Samples sequence is reducible check the checkbox to allow a runtime reduction of samples Drag the SMART Step Needle Wash Settings to the next line accept the default settings and click OK S
131. h em Ree eon E ES i ipette with Tips trom ML_STAR Smart Steps ee hove on VacuumBox 5 WE Error Handlir Instrument short name ML_STAR Aliquot i mE VacBox 1 Parallel Proc rt Fa Begin P Aspirate sequence ML_STASR Reagenst A p Ene Per Dispense sequence ML_STAR CooledSample controlling sequence ei PAL STR a rerneretetctetittetetttcntntntnnnntntnntintnieinsnananenenenenenestninannentntnnein l E careers 29 ML_STAR Smart YacBox 1 Duration 30 Pressure 5 Ramp 2 7 procedure Yolume per well Hl 50 Return value count 0 fn rt Load AB Pipette with EmmptyvWaste on WacuumBiax Prrere rete errr etter terete eter eee ae Unload eats ie VacBox 1 Duration 20 j Needle wast a Return value count 0 ae Needle Pick End Parallel 7 E E E E sca strse ter cards reer ter ire ret cer ceeded Cer tee a E a E cor iet T T restse tartar teraet ee ffl Needle Eject Yat for parallel process process1 with infinite 2 E en bd timeout 4 b J annn F Page 114 of 225 610766 02 Microlab STAR User Manual Let s discuss the individual steps of this method now Drag the parallel process icon A dialog appears to start parallel process No 1 Enter a name for the parallel process and click OK The method editor now shows the bifurcation of the normal and the first parallel process The STAR SW supports nested parallel processes Drag the Move Step of the vacuum box to
132. hange HAMILTON standard liquid classes store the class under a different name first For special applications the user can define his or her own liquid class to achieve the highest accuracy with the compounds and volumes of interest For this purpose a Liquid Editor comes with the Microlab STAR User Software It is described in Chapter 11 below NOTE Always use the same liquid class for one aspiration and dispense cycle Otherwise uncontrolled residual volumes may be left within the tip or other errors relating to the sum of volumes may occur 2 4 1 Liquid Handling Examples Here are some examples of frequently used combinations of liquid classes and pipetting modes 1 Aspirate gt 20ul of a water like liquid dispense 5 ul into an empty 96 well plate use standard tips change tips every cycle Liquid Class StandardVolume_Water_DispenseJet Aspiration Mode Aspiration Dispense Mode Jet Dispense Empty Tip Detection Aspiration LLD pressure or capacitance or both submerge to a depth of 2mm following liquid level Dispense Fixed height of 5mm not following liquid level 2 Aspirate a water like liquid single dispense into a pre filled 96 well plate use low volume tips change tips every cycle Liquid Class LowVolume_Water_DispenseSurface Aspiration Mode Aspiration Dispense Mode Surface Dispense Empty Tip in the liquid class selected here the blowout volume is 0 Detection Aspiration LLD Pressure or Capacitance o
133. having a 1 for each active and a 0 for each inactive channel e g 11100011 Having deactivated a channel the question arises whether to pipette all sequence positions with the remaining channels select 1 All sequence positions or to leave out the corresponding wells select 2 channel pattern Click OK to accept the settings Page 126 of 225 610766 02 Microlab STAR User Manual NOTE Before any other instrument specific step can be carried out the system has to be initialized Drag Load Carrier to the next line in the method For an instrument with autoload option this command loads the carriers automatically onto the instrument deck during runtime For a manual load instrument this command requests the user to load the carriers for runtime Load Carrier New Carrier to load 4 Advanced Error settings OF Cancel Help Specify the name of the carrier to be loaded The plate barcodes are stored under the default file name barcode_1 txt The positions on the deck are automatically retrieved from the deck layout on runtime Click OK Note that the checkbox barcode trace within the configuration editor must be checked to generate this file Repeat the Load Carrier command for the other plate carrier and the tip carrier To copy the whole source plate and not just the first 8 wells to the target plate the tip pick up aspiration dispense and tip eject steps have to be perfor
134. he Use Default checkbox A brief description of the error is given followed by the available recovery options Only one default procedure can be selected Among the choices are Cancel quits the current step and starts the user defined error handling if specified If no user defined error handling is present the method aborts Abort aborts the method Bottom Exclude Repeat Air The flag visible allows you to add the appropriate button to the error recovery dialog box The flag flag allows you to flag the error to the trace file Aspirate Default Error Handling Pressure LLD Error Ho Tip Error Execution Error Hardware Error Not Executed Error Syntax Error Improper Aspiration Error Clot Error Liquid Level Error M Use default Error description Liguid level not found or not enough liguid Exits default error handling user defined error handling is used if specified in method e ede ea eee a E eee ee dated anit Me Rese Bottom Repeat the step at the container bottom Timeout 5 3 Infinite Sound O M Abbrechen Hilfe Page 180 of 225 610766 02 Microlab STAR User Manual To enable walk away handling of errors disable the checkbox infinite and enter a timeout into the input field The runtime error dialog then pops up waits for the timeout and closes to continue with the default error recovery chosen for this error lf the user clicks o
135. he Microlab STAR Service Manual This manual will be made available to Hamilton authorized service technicians Whenever a manual amendment is issued detailed instructions on amending the existing manual will be provided 1 3 Intended Use of the Microlab STAR The Microlab STAR is a robotic pipetting workstation in other words a sampler used for pipetting liquid samples in an automated process suitable for medium to high throughput with a high degree of flexibility in pharmaceutical veterinary and genetics applications A user will typically wish to carry out low medium or high volume contamination free pipetting with disposable tips or with steel needles At the present time the Microlab STAR is classified as a general laboratory instrument and is not specifically validated as an in vitro diagnostic device 610766 02 Page 7 of 225 Microlab STAR User Manual 1 4 Operation The operator of the Microlab STAR must have attended an appropriate training course The procedures contained within this manual have been tested by the manufacturer and are deemed to be fully functional Any departure from the procedures given here could lead to erroneous results or malfunction 1 5 Safety The following section describes the main safety considerations electrical and biological in operating this product and the main hazards involved 1 5 1 General Precautions When using Microlab STAR Good Laboratory Practices GLP should be observed
136. he falling liquid level if specified to the bottom of the container staying there for the rest of the aspiration For dispensing four modes are possible e Surface Dispense Part Volume for dispensing only a part of the liquid in the tip toa surface leaving a residual volume in the tip e Surface Dispense Empty Tip for dispensing all the liquid in the tip to a surface e Jet Dispense Part Volume for dispensing only a part of the liquid in the tip in a jet i e without touching a surface leaving a residual volume in the tip e Jet Dispense Empty Tip for dispensing all the liquid in the tip in a jet 610766 02 Page 17 of 225 Microlab STAR User Manual The liquid class stores all relevant background parameters such as flow rates and volume corrections for one pipetting cycle i e for one aspiration and the subsequent dispense s Depending on the pipetting mode chosen only a subset of the parameters of the liquid class is active According to the different dependencies listed above liquid classes have attributes related to their intended use tip type liquid name and dispense mode Different liquid classes are provided with the User Software and optimized for different liquids tip types and important pipetting processes such as aspiration followed by dispensing either to a surface or in a jet HAMILTON has optimized the standard liquid classes with great care to assure the best pipetting accuracy To c
137. he tray i Holder for Carrier Click Load in the dialog and the carriers are loaded onto the deck automatically by the load carrier command in the method At the same time the barcodes of carriers and labware are read and stored in a file Alternatively load the carriers on to the defined positions of the autoload tray before starting the method Loading and barcode reading will then be performed without user input 2 To repeat what we said already if you have no LoadCarrier commands incorporated into your method you must load the carriers manually into the positions on the deck defined in the deck layout before the run is started Make sure that the carriers are inserted completely until they touch the rear connectors NOTE If no load carrier commands are specified in the method no check of carrier positions or correct loading is made Page 170 of 225 610766 02 Microlab STAR User Manual 15 Running the Microlab STAR 15 1 Running a Sample Method with the Instrument To run the method OnePlateToPlatePipette from the example section you need to access the run control Double click the Microlab STAR Run short cut on the desk top ky Microlab STAR RUN From the File menu of Run Control select Open and open the deck layout that was created for the method OnePlateToPlatePipette directory methods DemoMethods_MLStar Your method is now loaded Hamilton Run
138. hroughout this method to the variable TipCounterVariable as a string i e within quotation marks EU Assignment Edit Varable Value TipCountery arable MyStandardTips Cancel Help Define a user input request for the temperature of the TCC as well as the number of samples to be transfered e User Input Edit Dialog title input Retur value Sound El Timeout z I Infinite Input request s gt Variable Promt Tye Dean NoSamples a How many sampes t integer woe SS Bctem zi Carter Temperature ix Pia Add Hemose Cancel Help For every input a variable name a text to prompt for the variable within quotation marks the variable type and a default value have to be given Type in the inputs as shown in line 8 of the method overview Page 160 of 225 610766 02 Microlab STAR User Manual Set the TCC settings according to the following dialog Temperature controlled carrier Temperature control 1 On Temperature aa Time to temperature check B00 Go to nest step O Without waiting Error settings OK Cancel Help The temperature is now set to the variable CTemp the value of which is to be typed in by the user at runtime Now the tip counting is specified using functions from the tip counter library First the user will be requested to reset the tip counter if required oh Edit
139. ht 70 mm 610766 02 Page 187 of 225 Microlab STAR User Manual Click on the Segment 2 tab Segment Definition from container rim to base Segment segment 2 Segment approxiniation Cylinder Founded base seament C Rectangle C Y Cone base segment C Inverted Cone C W Cone 0 Upper internal Diameter li 0 mm Intermal segment Height mm a H f lt Back Cancel Help Select Round base segment and fill in the value for the upper inner diameter 10mm and the segment height 5 mm Click Finish to finish the container definition You are prompted for a name for the newly defined container e g choose MyContainer ctr and click save 17 3 2 Defining a Rectangular Custom Rack Now let s define a rack for the containers Ensure all the racks and containers you define have distinct obvious names for ease of use Page 188 of 225 610766 02 Microlab STAR User Manual Select New and Rectangular Rack from the File menu of the main window A series of query dialog windows starts giving the user the opportunity to design the rack Rectangular Rack Definition i Microtiter plate Portrait 86 127 mm i Microtiter plate Landscape 127 86 mml leon picture Browse Optional if an icon is selected it will be used for graphical display Back Cancel Help Choose always Rectangular Rack to define or m
140. ialog box appears Browse for the PLT_CAR_L5MD in the ML_Star directory Click OK to add the carrier Double click on one of the sites of the carrier Select Nunc_96_Flat_L rck from the Nunc directory Click OK Now you have added a 96 well plate to the carrier Double click on another track to add a tip carrier A dialog box appears Browse for the TIP_CAR_480 within the ML_Star directory Click OK Double click on one of the sites of the carrier Select standardtip_ rck from the ML_Star directory Click OK Now you have added a tip rack to the carrier Repeat this to add several tip racks to the carrier Save your deck layout under the name TubesToPlatePipette lay We will now define sequences which relate all samples on the one hand and all tip racks on the other hand Page 134 of 225 610766 02 Microlab STAR User Manual Under the Tools menu select Sequence Editor The Sequence Editor opens up a Tubes ToPlatePipette lay Hamilton Sequence Editor File Edit View Sequence Play Help S bed Bs 2 Q Q e in Labware IO Position ID 1 LT q lz lx Pick a comer Fick a direction Mo 4 dafa dbf Sort Racks A Sorthy Grid Columns A Sorthy Grid Rows A Filter by Probe Head For Help press Fl a Sequence Editor 610766 02 Page 135 of 225 Microlab STAR User Manual Zoom in by clicking the button on the tool bar Rubber band all four tub
141. ialog box has three main buttons Their functions are to load all sequences to the deck to add a specific sequence which can be selected from the dropdown field and to remove a selected sequence In the case of a tip sequence the SMART Step gives you the option to read the current position of a tip counter which has to be specified In this example the tip counter MyStandardTips is read and the current position of the sequence is set to the value of the tip Counter In addition sequences may be reduced on loading click on show details If a carrier for 1536 well microplates is being loaded it may be calibrated using the channel selected in the dropdown field Page 80 of 225 610766 02 Microlab STAR User Manual Click on Error Settings to define the error handling for this step This is similar to the error handling of the SMART Step Pipette To unload the deck use the SMART Step Unload It is very similar to the loading step lai Unload New ML_STAR kd E P A Here all carriers involved in the specified sequences are unloaded from the instrument deck 610766 02 Page 81 of 225 Microlab STAR User Manual 8 2 1 3 SMART Steps for Washing Needles For the washing of needles three SMART Steps are available The idea is that needles can be used like disposable tips without bothering about the organization of the three wash modules of a wash station To do this the needle pick up command waits
142. ibrayAHSLSeqhib st PY PBeaFiemoventt I Sequencechy WL STAR Hits a The parameter is the sequence ML_STAR Hits Page 148 of 225 610766 02 Microlab STAR User Manual Next the file holding the absorbance information is opened The file format is also defined within this step Drag the green Open File icon to the method tree tw File Open Edit Microsoft Access Microsoft Excel Structured Text File ASCII File All File Types File name xls 4deD ates xls Sheet name Absorbance a File handle Mode Open file to read Column specification J coumnname RW variabie Variable type _ Max colurnn width LabiD a foo e e dT Command string sqlselect Abbrechen Hilfe Select the tab for opening excel files In the first input field the file name is requested Note that the file type is an Excel file xls where the sheet name sheet1 if not defined otherwise and the sign must be added within the quotation marks Define a file specifier here the default file1 which is just a name for the file used within your method Data will later be read from this file making reference to this file specifier Select Open File to Read as a mode since you want to read data from the file Now define the file format Here a variable is assigned to each column of the file Later for each reading step one record one line is read from the file and the num
143. ick Next Type in the width and length of the rack here e g the outline of a microplate and the rack boundaries Click Next 610766 02 Page 191 of 225 Microlab STAR User Manual See section 17 5 for more information on these inputs Now the rack definition is finished Since we checked Load the Rack with Containers in the second screen we now have to define the containers which will be placed in the holes of the rack We have the choice either to browse the directories for defined containers extension ctr to fill our new rack with or to define a new container Container type Container type myContainer ct Browse Hew Distance from base of rack to mm base of container Container Offset M Use container type for all positions Advanced Back Cancel Help We now come to the end of rack definition with the new container pre selected Accept the defaults and click Finish to finish the rack definition Page 192 of 225 610766 02 Microlab STAR User Manual You now see your newly defined rack in the Labware Editor W4 Unbenannt Hamilton Labware Editor File Edit View Help 09 O O amp OO For Help press F Choose Save in the File menu to store the new rack under the name MyRack rck in the labware directory Select Exit from the File menu Once you are back in the Add Labware or in the Deck Layout edi
144. id if no rows are selected 4 AL i Ea Ex OOM oo0o000000 eoco0o0008 Sooo 0o0000 cocoo0o0eo0n0 eogoo0o000n eooo0o000n OOCOO amp OOOO OOM OOOO 6 costar Hat S woe IOoOOO00 oOo0000000000 OOOONOOOOOOOoo 4 af eel Sot Racks A Sorthy Grid Columns A Sort by Grid Rows Filter by Probe Head For Help press F Agesta flat ea HOt efe fietei ieT eoooeoocootl SOLOS LOIS Ee Lea eooo00oo00on0t BORSROLSLOIOLST booa booa Ki costar flat LS See eter sleleteleteleter E Slelelelelelerel 2 In this case the initial sequence was Rack1 A1 B1 C1 D1 E1 F1 G1 H1 Rack2 A1 B1 C1 D1 E1 F1 G1 H1 Taking the 4 row after the first row gives Rack1 A1 E1 Rack2 A1 E1 Rack1 A1 is gone so Rack1 B1 F1 Rack2 B1 F1 Rack1 B1 is gone so Rack1 C1 G1 610766 02 And so on Page 65 of 225 Microlab STAR User Manual 8 Graphical Method Editor Methods are short programs that string a number of specified commands such as aspirate or dispense together to instruct the instrument to perform a function They can be as complex or as simple as the user desires Access to a method editor is necessary to define or edit methods The Graphical Method Editor is a syntax free editor that presents a batch like graphical editing environment Common constructs such as loops conditionals and built in functions are supported by this editor It also provides a variable management system Thi
145. iers mentioned in the upper part of the dialog box the 2 plate carriers and the tip carrier Don t forget to place labware onto the correct positions Autoload Only Whenever the system finds a Load Carrier command in the method the user is requested to feed the carrier holding the appropriate labware onto the autoload tray The correct position is highlighted by LEDs Alternatively all carriers can be placed directly in their correct positions on the autoload tray Click OK in the dialog box to start loading Page 172 of 225 610766 02 Microlab STAR User Manual At the end of the method you are requested to unload the carriers from the deck The following dialog box opens Unload Sequence Iofs The following carriers will be unloaded tip car 480_ 0001 plt_carlSmd_O002 plt_carl5rmd_O001 Heset RemoveAll je je Help Manual Load Only Click OK within the dialog Unload all the carriers manually The unloading will be checked by the system Autoload Only Click OK within the dialog The carriers are unloaded to the autoload tray 610766 02 Page 173 of 225 Microlab STAR User Manual The method is now finished Within the method trace file the method completion information is visible fee OnePlateT oPlatePipette_lay Miel x 1310 02 Microlab STAR UnloadCarrier start 13 10 02 Microlab STAR UnloadCarrier complete gt plt_carlSmd_OQU0 Single Step 13 10 03 Microlab STAR UnloadCarner
146. in You now can continue or abort method execution To abort the method click abort You will be prompted to confirm the abort NOTE An abort may cause the loss of data Note that aborted methods cannot be restored again A fast abort can always be done by opening the front shield of the STAR during run execution 15 2 Run Simulations It is also possible to run a simulation instead of the instrument It is recommended always to simulate a newly created method first before running it on the instrument The run simulation is switched on in the configuration editor Access the configuration editor from the run control by clicking on the deck layout frame Only now is the Tools menu visible Select configuration editor Hamilton Microlab STAR Configuration Editor Select mode for instrument Microlab STAR About C Instrument Help Save amp Exit Select Instrument Exit Set the switch to simulation FETE NOTE FOR MANUAL LOAD INSTRUMENTS Make sure the autoload checkbox is checked for the simulation Click on advanced and Select the tab instrument configuration Check the autoload checkbox To run the instrument set back the configuration On loading commands the simulator responds for each carrier to be loaded and unloaded with the message LoadCarmer Deck position 1 to 6 i not free Is the carier on this position s the night one Click Yes to continue
147. in chapter 2 The Art of Pipetting page 12 the background parameters for pipetting are managed using liquid classes A liquid class is a set of parameters specifying the aspiration and dispense behavior appropriate for a given liquid e g water DMSO In all aspirating and dispensing steps a valid liquid class must be selected Standard liquid classes will be supplied along with the User Software Water DMSO Glycerine Precinorm The standard liquid classes cover a wide range of applications and you will probably not need to make any changes to the parameter settings For special applications you can define your own liquid class This custom liquid class can be used just like the pre defined classes Defining liquid classes independently serves to simplify the steps in the method and allows the complete set of parameters to be defined once for all pipetting tasks To define a custom liquid class and to display the parameters of the liquid classes defined the Liquid Editor should be used 11 2 Editing Liquid Details To start the Liquid Editor select ML_STAR Liquid Editor from the Tools menu of the Deck Layout Editor The Liquid Editor start window is displayed fe Microlab STAR Liquid Editor C Programme HAMILTON Configi ML_STARLiquids mdb O x File Liguid Class wiew Window Help Liquid Classes fel Liquid Class Name StandardvolumeFilker Serum Aliquotjet 1 0 Serum For aliquot a volume tip w Standard
148. including extension Create sequences for all tips AllTips and for all tubes AllTubes using the sequence editor Open the method editor and link the deck layout to the method to be written Link the following libraries to the method method gt libraries Libraries Add Library Remove Library cared Hob 610766 02 Page 157 of 225 Microlab STAR User Manual NOTE This method uses the sample tracker Make sure the checkbox sample tracker within the configuration editor is checked The name of a method using the sample tracker must not exceed 20 characters including the extensions Finally the method should look like this following the first step Initialize the display of method steps continues overleaf EA Example med Method LoadCarrier on ML_STAR i Ji Rack type TempCarrier Barcode file barcode_1 txt Return value count 4 Loadtarrier on ML_STAR Pack type tubes Barcode file harcode_1 txt Return value count 4 LoadCarrier on ML_STAR er Rack type tubes Barcode file barcode 1 txt Return value count 4 LoadCarrier on ML_S TAR Wack type tubes Barcode file barcode 1 txt Return value count 4 Assignment Tiplounter ariable MyStandardTips Comment Set TLC User Input Dialog Title Input Return Value Sound Timeout nfinite Input MoSamples How many samples today Integer 96 1 96 CTemp Ca
149. iration Mode Dispense Mode Detection 610766 02 StandardVolumeWaterAliquotJet Aspiration Jet Dispense part volume Aspiration Capacitance LLD Submerge Depth 2mm following liquid level Dispense Fixed height of 5mm not following liquid level Page 19 of 225 Microlab STAR User Manual The following table gives sample values and results for pre and post aliquot volumes please note that these are not technical specifications Chan Tip Liquid Pre V pl No of V ul V yl CV R Class nel Type wet main Alig pre post Table of Aliquots Tip types are Std Standard Volume Tip 300ul High High Volume Tip 1000ul Pre wet If Yes pre wetting by 3 fold mixing on aspiration with aspiration volume necessary V main alig Volume of main aliquot V pre alig Volume of pre aliquot V post aliq Volume of post aliquot CV Precision coefficient of variation for definition see specs R Trueness for definition see specs The R and CV values mentioned here are typical results for measurements Class Liquid class to be used A StandardVolume_Water_AliquotJet B StandardVolume_Serum_AliquotJet C HighVolume_Water_AliquotJet D HighVolume_Serum_AliquotJet The dispense mode for all cases is Jet Dispense Empty Tip 2 5 Process Control 2 5 1 Monitored Air Displacement The Microlab STAR is equipped with an aspiration monitoring feature During the aspiration
150. ircular racks are specialized grids for holding either CO RE tips or containers in a segment of a circle The filename has the extension crk 17 3 Example Defining a Rectangular Rack with Containers The following sections illustrate the procedures for defining labware using the example of a rectangular custom rack and containers 17 3 1 Defining a Container Let s define a new container At the end of the container definition we can use that container in a rack definition To start 1 from the labware editor select New and then Container out of the File menu or 2 inthe Rack definition dialog press the New button 610766 02 Page 185 of 225 Microlab STAR User Manual In both cases the following dialog box is shown Container Definition Number of segments E T Container clearance height rel to base fen mm Masimum pipetting height rel to basel rr Touch off at bottom height rel to base 0 3 mm Thickness of container base li mm W Liquid level detect valid Liquid seeking height rel to basel 75 rrr T wick side of container valid loony picture Browse IF an icon i selected it will be used for graphical display Anal Options Back Cancel Help We define the containers as round bottomed tubes of diameter 10 mm with a total length of 75 mm Indicate the number of container segments here 2 because our tube has a cylindrical and a round bottomed part
151. is ML_STAR Plate and not reloadable LLD settings are as in the previous exampes cLLD on aspiration Sensitivity low and fixed height on dispense The dispense mode is now Jet can t be changed The liquid is Water for Aliquot 610766 02 Page 141 of 225 Microlab STAR User Manual Click OK to accept the settings Click OK to quit the summary Your method is now ready to go 13 6 Cherry Picking or How to Change a Sequence Within a Method Using SMART Steps Assume you have a source plate and a photometer reads the optical absorbance of the wells of the plate You now want to create a target plate with all the compounds in the source plate having an absorbance of A gt 1 0 The cherry picking method does exactly this The photometric results are retrieved from a file and a sequence of hits A gt 1 0 is created on the fly according to the absorbances read Pipetting then occurs according to this sequence This method does not use Load Carrier commands The system therefore expects the carriers to be loaded manually onto the deck at the defined positions before the run is started The method can however be easily adapted for automatic loading with autoload option by inserting the Load Carrier commands after the Initialize command For this method we need a database containing the absorbances of the 96 wells of the source microplate The database can be an ASCII text file a Microsoft Exce
152. is equipped with a front shield and a hinged transparent window made of plexiglass This window is equipped with a magnetic switch and it is locked during a run AN ATTENTION An aborted run stopped by opening the front window cannot be restarted If you need to open the window during a run click Pause in the Run Screen wait until the instrument stops and then open the window 3 2 Options Options are defined as components or configurations that are part of the instrument initially provided to the customer as specified by that customer Options cannot be added later whereas accessories can see Section 3 3 Accessories below The instruments configuration is set within the configuration editor of the User Software 3 2 1 4 8 12 or 16 Pipetting Channels The Microlab STAR comes with 4 8 12 or 16 pipetting channels working in parallel for simultaneous transfer of liquids Instruments with 4 8 or 16 channels are best operated with carriers holding microplates and tip racks in landscape orientation e g PLT CAR L5MD whereas carriers with portrait orientation for microplates and tips e g PLT CAR P3MD are best suited for the 12 channel Microlao STAR In addition increasing the number of channels to 12 or 16 reduces the random access space of the Microlab STAR i e the space that can be reached with all pipetting channels This is shown in the following table No of Yorn Imm Ymax fmm Channels Pet to str Coor
153. is specified that exceeds the volume of the tip multiple transfers with equally divided volumes will be performed automatically A residual volume may be used too Then the choice is to discard the residual volume back into the aspiration sequence or to dispense it into the waste container In the case of aliquoting a pre aliquot has to be specified too This pre aliquot is dispensed back into the aspiration sequence A redispense height counted from the bottom of the container has to be given Now the aspiration and dispense locations used within this step have to be defined Dispense sequence is controlling Dispense sequence MLS TAR MyPlate C Reload iM Reducible by user Multiple use Aspirate sequence is controlling spirate sequence MLS TAR tubes C Reload i Reducible by user f Not reload I Aspirate al LLD Advanced aa LLD Advanced i Jet C Surface Liquid liquid class The sequence of the aspiration and dispense is to be selected from the dropdown fields In addition the user can opt to reload a sequence if during pipetting a sequence is used up no more tubes are present to aspirate from the system then asks for a new carrier of tubes to be loaded If in addition the checkbox reducible by user is checked then the user has the option of reducing the newly loaded sequence at runtime This is especially helpful if the exact amount of sample tubes varies from run
154. ition ofa Carrier position high density microtiter plate channel before aspirating or dispensing MoveTo Moves the pipetting head to an Mode direction value Position absolute position or to one relative to the current position Firmware Sends a low level firmware Firmware command Command command to the instrument parameter Get last liquid Gets the z position of the last level liquid level detection Start Needle Starts a needle wash module Name of wash module Wash Wait for Wash Waits for the needle wash Name of wash module module to be ready 610766 02 Page 87 of 225 Microlab STAR User Manual Command Icon Action performed Set Sets the temperature of a TCC Temperature Get Retrieves the temperature of a Temperature TCC Get Plate Picks up a plate from the defined position Place Plate Sets a plate down in a defined position Move Plate Transfers a plate to another sequence Spreads the fingers of iSWAP s Open Gripper j i robotic hand Close Gripper Closes the fingers of iSWAP s robotic hand NOTE Parameters to be specified Carrier name temperature settings Carrier name bind return value to get temperature in C see chapter about binding return values Position of plate to be located and picked up by iSWAP Movement type transport mode plate sequence sequence counting lid sequence complex movement parameters Move to sequence Transport mode plate sequence s
155. itor is used to switch between simulation and instrument mode NOTE To simulate a run always set the instrument configuration to autoload even for a manual load instrument Don t forget to set the configuration back to manual load when performing a run with a manual load instrument 610766 02 Page 99 of 225 Microlab STAR User Manual 10 1 Method Linking The Microlab STAR user software allows you to link different methods together prior to starting the merged or linked run To do so open the Deck Layout Editor and select File gt Method Linking A dialog appears Method Linking ki File Djaj Deck layout files Execution sequence SSS HTB hal Hel 75 sl Append gt He 14 hel Hes Hal H439 hal Insert gt Hed hal Hed hal Hx43 hil Remove s Exit Select the first method of interest and click on the buttons to add the method to the list on the right hand side It is recommended to select HSL files hsl rather than layout files lay or method files med Select File gt Save to generate a chained method file cmt Close the dialog by clicking on exit Close the deck layout editor Open the Run Control by double clicking the shortcut on the desktop Open the newly created cmt file and execute it Page 100 of 225 610766 02 Microlab STAR User Manual 11 The Microlab STAR Liquid Editor 11 1 Concept of Liquid Classes As mentioned
156. ked red upper and left most well A1 610766 02 Page 57 of 225 Microlab STAR User Manual 6 3 Making Changes to the Deck Layout It is possible to add additional labware to the deck any time by the procedures described so far To delete labware items from the deck right click the item to be deleted and select Delete from the menu In the message box that appears click OK The item is deleted from the deck To rename a given labware item right click the item to be renamed and select Rename from the menu In the message box that appears change the name and click OK The item on the deck is renamed To get the properties of a labware item right click the item of interest and select Properties from the menu The message box that appears will display the type and the name of the item For the definition of custom labware objects refer to chapter 15 NOTE If you change a name or delete a labware item on the deck which is already used in a method you must also make the appropriate changes to your method Page 58 of 225 610766 02 Microlab STAR User Manual 7 Sequences A sequence is a defined series of pipetting events involving the transfer of liquid from one container to another A sequence defines the order of containers in a rack For example it links the single wells of a microtiter plate together so as to treat them as one plate When the probe is pipetting it will follow this order channel No
157. l file or a Microsoft Access database In this case our database is an Microsoft EXCEL file containing three columns The LabID defining the plate name of the source plate Source_1 for the first plate and Source_2 for the second plate the PosID defining the position within the microplate alohanumerically A1 A2 H12 and the absorbance or optical density OD The work list therefore has193 lines 1 header line and the entries from 2 plates with 96 wells Page 142 of 225 610766 02 Microlab STAR User Manual w Microsoft Excel AdsData3 xls 0 x 5 Datei Bearbeiten Ansicht Einf gen Formak Extras Daten Fenster x ies LabID FPoslOD OD 2 Source 7 Al 0 506264 3 Source_1 B 0 956173 4 Source Ci 1 010817 5 Source_1 D1 1 127001 B Source_ E 1 010192 7 Source_l Fi 0 487826 8 Source_l G 0 996693 9 Source_1 H 0 664385 10 Source_ A 0 83523 11 Source_1 B2 0 834056 12 Source C2 1 094375 13 Source_1 D2 0 46083 14 Source E2 1 024097 15 Source_1 F2 0 563819 a HE Saure 112 bd Mla kH Absorbance fF AT rT f 4 The work list ADSData3 xlIs The name of the sheet is Absorbance Double click on the text Sheet1 within your Excel file and edit it to Absorbance see the section on working with Microsoft Excel The deck layout looks like this standardtip eTtt t Hanun a5 fi ET For this method place two plates called S
158. late manufacturer The ramp duration is the time needed to reach the desired vacuum This value may need to be determined by trial and error The vacuum duration is the length of time that the desired vacuum is to be applied Variables can be used for Vacuum box number Ramp duration Vacuum duration and Vacuum The vacuum pump will operate for a time equal to the ramp duration plus the vacuum duration In the event that the desired vacuum is not achieved during the ramp period an error will appear during run time Automatic error recovery is possible by defining recovery options in the method If you click the Error Recovery button in the Vacuum Parameters dialog box the Vacuum Error Recovery window appears Error recovery option in the case of not reaching desired vacuum within the specified ramp period f Invoke user recovery ignore and continue Abort OK Cancel Help If you select Invoke user recovery which is already selected as default the operator can decide what to do at run time Ignore and continue and Abort which will release the vacuum are the other possibilities After selecting an error recovery option click OK 610766 02 Page 113 of 225 Microlab STAR User Manual 12 3 Parallel Processing with the AVS We now discuss a sample program to illustrate the use of the vacuum box software O x File Edit View Tools pelaj S tale Bla Help
159. le barcode_1 txt Return value count 4 Comment Copy plate gt over folowing Sequences ML_STAR Source joopCountert used as loop counter variable TipPickUp on ML_STAR Channel 1 11111111 Optimized channel use All sequence position Sequence ML_STAR Tips Sequence counting 1 Automatic Return alue count 3 Aspirate on ML STAR Channel 1 11111111 Optimized channel use All sequence position Sequence ML_STAR Source Sequence counting 1 Automatic Liquid name Standard olume_ Water DispenseJet Volume 50 ul Mix volume ul Cycles 0 Position relative to liquid surface mm LLD settings on Capacitive 3 Return value count 3 Dispense on ML_STAR Channel 1 11111111 Optimized channel use All sequence position Sequence ML_STAR Target Sequence counting 1 Automatic Liquid name Use the same liquid as aspirated Yolume 50 pl Mix volume pl Cycles 0 Position relative to liquid surface i mm LLD settings off Liquid tollowing om Return value count 3 ipEject on ML_STAR i Channel 1 8 11111111 Optimized channel use All sequence position sequence ML_STAR VWvastel6 sequence counting 0 Manually Return value count 3 End Loop ML_STAR Source Reset Sequence after loop co pe ee Comment Unload deck Unload arrier on ML_STAR Rack type tip_car_460_0001 Return value count 3 Unload Carrier on ML_STAR Rack type pt_car_lSmd
160. lt d2 _6 _costar_flat_l2 430 _6 _costar_flat_lt 6 costar flat_I2 6 costar flat 6 costar flat 2 mn tae inateretelatere BORO OR EROLOLOLOT OL STS george e00000o00n8 george e00000o00n0 ervoee0000o0n8 ah 6 t flat ie _ age 06 ROBOT eLOLO ROR geooo00o0r ooo00o00e eo 0000000000 ooo0c0o0oc eo 0000000000 ooo0o0o0oc e o0000000000 oo0o0c0o00c 5 tar _flat_ls 5 tar_flat_l4 E WETEWOOOOOOO WUE sac L SO O009C000 a0 OIO000 0c CIC CIO Caco Coo coe 0202000 4 d x Pick a corer Pick a direction gt KAA __ o Serpentine 4 ila b LF Sort Racks A Sorthy Grid Columns A Sorthy Grid Rows A Filter by Probe Head For Help press F1 Page 64 of 225 610766 02 Microlab STAR User Manual 7 1 8 Sorting Grid Rows Example The same initial sequence if sorted using Sort by Grid Rows will result in the following a metl lay Hamilton Sequence Editor Eie Edt View Sequence Play Help e amp 2 BQ Q e F Vfinew J tabware 4 _6_costar_flat_lt Z _6 costar_flat_lt _6_costar_flat_l2 4 _6 _costar_flat_l2 _6_costar_flat_l 6 _6 costar_flat_lt 6 costar flat 2 E E costar_flat_l2 9 E costar_flat_l _6_costar_flat_l _6_costar_flat_l2 6 costar flat 2 _6_costar_flat_lt 6 costar flat 6 costar flat 2 _6_costar_flat_l2 Coalesce every 4 rows in grid starting at row fi from top of selected rowe or from top of gr
161. m System As a first step to writing the method a deck layout with an integrated vacuum box has to be created Therefore open the Hamilton Deck Layout Editor window and select New from the File menu This window appears l OK Instruments Cancel Vacuumb ox Select the Microlab STAR instrument and click OK An empty STAR deck layout will appear Page 106 of 225 610766 02 Microlab STAR User Manual You have now to select the vacuum box under Add labware from the labware folder VacBox under MLStar and put it in the right place on the deck track 31 for first vacuum box TUnbenannt Hamilton Deck Lay 0 x Fie Edit view Tools Help Oea S ae aa e Edit Method E i Run Method 5 10 15 0 25 30 35 40 45 K For Help press F1 B Tr 1 000 i 34 2 To add a filter plate on the conditioning site of the vacuum box double click on position 2 of the vacuum box and select the right filter plate The same filter plate can be selected for the elution site of the vacuum box on position 1 On the same STAR deck layout all required carriers can be added with the Add Labware command under the Edit menu Finally select Save from the File menu to save your deck layout with the integrated vacuum box 610766 02 Page 107 of 225 Microlab STAR User Manual There is also a possibility to permanently change the default STAR deck layout which might
162. med 12 times This can be achieved by a loop Drag the Loop command to the next line of the method window The loop statement consists of two lines a begin loop and an end loop statement Whatever code is inserted between these two statements will be looped 610766 02 Page 127 of 225 Microlab STAR User Manual The loop dialog window looks like this 28 Loop Edit terate a fixed number of times Number of iterations H terate while expression is true Lett operand Comparison operator Right operand X equal ta O e O esee e hl STAR Wasted A terate over files a Opened Files Loop counter variable loopCountert Cancel Help A loop can be performed looping over a fixed number of iterations over an expression repeat while the statement in the expression is true a Sequence or a file until the end of file is reached Here we loop over the source plate sequence This means that the loop will continue until all sequence positions the 96 wells of the source plate have been used Then the loop will stop Choose the default after loop for the Reset Sequence option to reset the sequence source to the initial position 1 after the loop is done If you pipette later to the same sequence source the sequence will then start at the first well again NOTE Keep in mind that if you loop over a sequence the sequence has to be incremented within the loop
163. n be removed Positions can be reordered as described above The sequence can be resaved to fix its positions according to the new order The tree view in the left pane displays the sequence from a tree oriented perspective where the tree s leaves are the positions in the sequence ralph lay Hamilton Sequence Editor 0 x ae amp Fa lAQ Ne Eile Edit View Sequence Play Help mimi m Em fa a Ea Aaa mim m Em fa a Aa Aaa oo00cooo mimi m Em fa Ean Ean Aar Number of rows of tips fi Number of columns of tips E gt m a Distance between each row E mmo Distance between each columr E mm Tipi row number f Tip 1 column number f1 y Serpentine Tip indexing Hie Defautts Increment tip indeces fram row 1 by C Row Column T Serpentine while incrementing tip indeces HACA For Help press F1 Sort Racks Sort by Grid Columns Sort by rid Roms Filter by Probe Head 7 1 1 Adding Positions to a Sequence Use the mouse to select labware positions to insert in the sequence as follows 1 Left clicking on labware position adds the position to the sequence That means it either a inserts it in sequence ahead of the first selected row in the grid or b appends the position after the last row in the grid if no rows are selected 2 Rubber banding a rectangle of wells adds a block of positions to the sequence The block of positions is added in a manner similar to
164. n the error dialog during the timeout the walk away will be stopped and the user has to select a recovery and continue manually For a list of all errors and their recovery options refer to the online help and the error settings dialogs However the most important errors are the ones listed in the foregoing chapter 610766 02 Page 181 of 225 Microlab STAR User Manual 16 The Microlab AT Barcode File Filter Along with the software comes a filter tool to generate a Microlab AT like barcode file This filter can be started manually or from the shell command of a method see example The sample tracking works as follows Check the check box sample tracking in the configuration editor At runtime one Access based data base mdb will be generated within the logfiles directory storing all liquid transfers of one method An additional register file HxRunIndex mdb stores information about all runs performed After the method or at least the liquid transfers are finished the filter tool can be called to generate the Microlab AT like barcode file from the data base The filter tool is stored under hamilton bin NOTE To run the AT filter tool the method name must not exceed 20 characters including extensions The plate names used in the method must not exceed 20 characters Given that a method has been run with the sample tracking enabled and a 96 well plate has been prepared the directory c barcodes now cont
165. nd z gt Timeout z I Infinite Input request s gt gt O Variable Prompt Type befaut 4 threshold Enter the threshold f Add Renove Cancel Help The dialog title is the text on the blue top bar of the dialog The variable name is threshold the text for the prompt is Enter threshold for hits within quotation marks the variable type is string and the default value is 1 0 within quotation marks All other input fields are optional Within the next step a calculation is made with the string variable read from the user input BA Assignment with Calculation Edit Var able Expressiorr SELECT FROM Absorbance threshold Cancel Help The variable sqlSelect is also a string It is generated by adding the sql selection statement and the threshold value This statement if applied to the data set selects only the records lines having absorbance values A gt threshold 610766 02 Page 147 of 225 Microlab STAR User Manual The syntax of the SQL statement is SELECT FROM absorbance WHERE OD gt Here absorbance refers to the name of the Excel sheet which is going to be opened during the next steps Now the sequence Hits as generated within the sequence editor is reduced to 0 all entries are deleted using a library function call SeqHemoyveall of HSLSeqgLib Edit JE Programme HAMILTONSL
166. nePlateT oPlate File Edt View Insert Tools Window Help _ o D eS tS oo e Awan X P ew device ml_star HSL OnePlateToPlate lary J Do not insert text before this line 7 Aspirate Calibrate 1536Plate oo TransterPlate UnloadCarrier m x mal 4 Syntax Chech HSL Method Editor Save the method to be created under the name HSL_OnePlateToPlate hsl Type in the first line of our method device ml_star HSL_OnePlateToPlate lay This command loads the Microlab STAR device driver and links the device to a concrete Microlab STAR deck layout context and the associated instrument The steps in the method we are going to write will appear between the curly brackets of the method skeleton One HSL program must contain exactly one such method We enter a method name by replacing oneplatetoplateMethod method TransferPlate As a first step in the method we initialize the Microlab STAR Under the sub menu ml_star drag Initialize to a line between the brackets A dialog window appears Click OK to accept the default values The line in the programming window should now read ml_star Initialize Channel 1 8 oo000000 If it reads only ml_star Initialize select View all step data in the View menu The parameters of the command are given between the parentheses Here all channels are initialized the circles giving a graphic representation of the situ
167. nel diameter of 9 mm If the system is paused do not wait too long before resuming the run Loss of liquid from a full tip may result in invalid data Do not overfill reagent containers tubes or other liquid containers Switch off the power of the Microlab STAR during maintenance Set up the Microlab STAR with its back to a wall to preclude any rear access to the work area The breakdown of the power supply during a run may cause the loss of data If the loss of data is unacceptable use an independent power supply Page 8 of 225 610766 02 Microlab STAR User Manual Opening the front window during a run will lead to a system abort and may cause the loss of data To avoid computer breakdowns ensure that there is always enough storage capacity on your hard drive Never disable any security measure Do not leave tips or needles picked up on the pipetting channels for long periods such as overnight This may cause damage to the O rings For repair or shipment all mechanical parts must be put in their rest positions A Microlab STAR sent away for repair must also be decontaminated if it was in a laboratory environment with infected or hazardous materials The Microlab STAR must be repacked in the original shipping crate and only by an authorized service technician There should be no containers or tips on the Microlab STAR during transportation Only original HAMILTON Microlab STAR specific parts and tools may be used with the
168. ns and field verification specifications are given too The trueness and precision specification is valid for 8 channels The basis for the calculations is given in Appendix E Principles of Calibration Page 40 of 225 610766 02 Microlab STAR User Manual The design specifications mentioned above are valid under the following conditions obtained for measurements at HAMILTON Bonaduz e Test method Gravimetric testing at Hamilton The scatter of the test method must be less than 1 6 of the specified precision for one channel e Trueness Precision The values given refer to use of 8 pipetting channels e Test size gt 12 single pipettings per channel with disposable CO RE tips pick up and dispense tip used only once per channel and specitied volume e Test mode Volumes 20ul as jet dispense lt 20ul as liquid surface dispense e Acceptance criteria Measured values are within specifications if less than the values appearing in the table on p 40 e Testtemperature 2042 C e Test fluid Deionized water with 0 1 NaCl 0 01 Tween NOTE No warranty can be given that the above specifications for trueness and precision are met with any other liquid or environment than the ones specified Optical test methods such as fluorescence or absorbance plate reading tend to have internal scatters in the range of about 5 The operating temperature range for the Microlab STAR is from 15 to 35 C with a relative humidity of 3
169. ns in the active sequence The order is applied to the currently selected labware positions or to the entire sequence if no positions are selected Use this filter to create processing sequences across multiple racks in the direction required l lx Pick a comer Pick a direction r __ l Serpentine Sort laf a D Sort Racks A Sort ky Grid Columns f Sort by Grid Roms A Filter by Probe Head 7 1 4 Sorting by Grid Columns The Sort by Grid Columns dialog allows you to sort the contents of the grid based on specified criteria Left most combo boxes contain the grid s column headers Labware Position X Y Z and none Right most combo boxes contain ascending and descending The sort is applied to the currently selected labware positions or to the entire sequence if no positions are selected l le Sort b i Y Then by Altemate ascending descending x Instruments Then by Altenate ascending descending eaa System Sort alaj bf Sort Racks A Sort by Grid Columns A Sort by Grid Rows A Filter by Probe Head f 610766 02 Page 61 of 225 Microlab STAR User Manual 7 1 5 Sorting by Grid Rows The Sort by Grid Rows dialog allows you to sequentially order every i th row in a grid beginning with the j th row The sort is applied to the currently selected labware positions or to the entire sequence if no positions are selected The sequen
170. nt to an ML STAR will result in an error message 8 2 1 SMART Steps SMART steps are powerfull commands for programming the Microlab STAR Whereas single steps see next chapter are restricted to single actions like picking up tips aspiration dispense etc SMART steps combine many single steps for specific tasks like filling a microplate starting with tubes aliquoting of reagent to a complete plate loading the deck etc The available SMART steps are Icon Command Action performed Parameters to be specified Sequence carrier Load Deck Load carrier on deck calibration Pipette With et oh pi pette mode volumes asp disp Tips Pipette peu disposable ups control seq liquid class tip seq tip handing replacement tip counter Unload Deck Remove carrier from deck Sequence Define Needle Specify parameters for needle Wash sequence start Wash wash with liquid settings Pick up Pick up Needles from Wash N eedle sequence Needles Station or Racks qu Release Release Needles in racks or l N le sequence Needles wash station and start wash ihe 1 Pick up tips Pick up disposable tips from tip Tip sequence tip rack counter Eject tips Eject disposable tips into tip none waste The method editor allows you to freely combine SMART steps and single steps For all SMART steps the instrument name is to be selected This is set by default to the Microlab STAR s short name and cannot be changed within the SMART steps Only in the case
171. ode and register files Page 46 of 225 610766 02 Microlab STAR User Manual 5 Instrument Configuration As described in Chapter 3 the Microlab STAR is available in several variants To configure the User Software to a specific instrument configuration the Configuration Editor is available within the Deck Layout Editor under Tools gt Configuration 9 Hamilton Microlab STAR Configuration Editor Select mode for instrument Microlab STAR About C Instrument Help Save amp Exit Select Instrument Exit ETE The first decision to be made prior to a run is whether to run the method with the instrument or to simulate the run without using the instrument Run simulation is a tool to test whether a method has been programmed correctly The run simulator creates a method trace file with pipetting pattern and other useful information Click the Advanced button to access the instrument configuration screen Different tabs are visible Instrument extensions Error settings Step selection Instrument configuration Barcode settings Instrument configuration Number of channels Default waste i Channel type 300ul Channel type 1000u MW Autoload Teaching Mode Teaching box i Kevboard C Both Teach with channel na E The tab Instrument configuration Select your instrument configuration Waste16 as in picture above is the correct sequence of the waste container for disposa
172. odify rectangular racks and microplates Choosing microtiter plate requests only a subset of the information relevant for the ML STAR Click Next Rectangular Rack Definition Number of rows J Use probe to measure rack iW Include the rack boundary Holes per row 4 M Load rack with containers Hole inner diameter 12 mm Stack height 0 ack heig mrm i Clearance height 100 mm Stagger Inside f None F By 0 mm C Outside Zuriick Abbrechen Hite Specify the number of rows here 2 the number of holes per row here 4 and the inner diameter of the hole here 12 mm which will later receive the container Check Load Rack with Containers and give the overall clearance height of the assembly here 100 mm Also check Include Rack Boundary to allow the boundaries to be set in the next steps Accept 610766 02 Page 189 of 225 Microlab STAR User Manual the default for Stagger an option allowing you to shift the rows with respect to each other and click Next The next dialog appears R ectangular Rack Measurements irae ee eed ee Specify the distance between the holes in a row here 20 mm and the distance between the rows here 30 mm Click Next Page 190 of 225 610766 02 Microlab STAR User Manual Rectangular Rack Indexes Accept the defaults for the indexing of the holes for alpha numeric indices ranging from A1 to H12 and cl
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174. of 225 610766 02 Microlab STAR User Manual 14 Loading the Microlab STAR 14 1 Manual Load To load the Microlab STAR first load the carriers with the appropriate labware 1 If you have incorporated LoadCarrier commands into your method you can start the run and the instrument will prompt you to load the carriers manually onto the deck Make sure the carriers are inserted completely until they lock into the rear connectors A magnetic detector checks whether your loading of carriers is correct Magnetic Sensor 2 If you have no LoadCarrier commands incorporated into your method you must load the carriers manually into the positions on the deck defined in the deck layout before the run is started Make sure that the carriers are inserted completely until they touch the rear conneciors NOTE If no load carrier commands are specified in the method no check of carrier positions or correct loading is made 610766 02 Page 169 of 225 Microlab STAR User Manual Autoload When using the Autoload option first load the carriers with the appropriate labware 1 If you have incorporated LoadCarrier commands into your method start the Run Screen as described in the following sections and run the method The correct positions for the insertion of carriers will be highlighted by the green LEDs Insert the carriers into the tracks of the Autoload tray until they touch the holding pins on the far side of t
175. of two or more instruments linked together is a selection necessary Page 70 of 225 610766 02 Microlab STAR User Manual 8 2 1 1 SMART Step Pipette with Tips The SMART step pipette is a multifunctional command for various kinds of pipetting tasks The Pipette Step allows the user for example to copy one plate into another or to transfer from tubes to a plate with variable numbers of samples or to aliquot reagent into a plate all within one step without taking explicit care about the instrument s initialization single steps for aspiration and dispense sequence handling or loops As is the case for all Microlab STAR user software components SMART Steps too make use of sequences Therefore the pipetting task should be reflected within the sequences defined for this step For the explanation of sequences we refer to chapter 7 In this chapter the general features of the SMART Step are explained examples can be found in the chapter Methods for Microlab STAR Dragging a SMART step into the method tree opens the SMART Step input dialog MHH Pipette with Tips Hew General Instrument short name Tip sequence ML STAR ka MLS TAR tips_ 1 Standard pipette procedure Aliquot procedure W Reducible by user Pipette mode i Simple 1 1 E Salei Multiplicity ri Pooling n 1 Tip handling replacement After each dispense One tip set E After fanstered volume C None i After ea
176. oices are available some of which are deactivated e New deck layout e Open deck layout e Save deck layout e Method editor e Run execution e Labware editor e Sequence editor e Zoom in deck layout e Zoom out deck layout Page 50 of 225 610766 02 Microlab STAR User Manual 6 1 New Deck Layout Under the File menu or in the toolbar select New to start the definition of a new deck layout If multiple instruments are installed on your PC a dialog window will appear Select an instrument e g ML_STAR for Microlab STAR and click OK If there is only one instrument installed the template base definition of a deck layout of that instrument is shown directly Select Instrument Instruments Cancel ML_SWiAP1 400 ML_SWAP420 Select Instrument window The deck layout displayed below shows the tracks of the predefined grids from the appropriate deck template i Unbenannt Hamilton Deck Layout Editor File Edit View Tools Help Dea Eje lS dy Edit Method Waste Run Method 5 10 15 i 245 30 35 40 45 50 For Help press F1 be 614 000 5 Example The Tracks of a Microlab STAR deck Use the zoom button to enlarge the deck view A deck template is an instrument specific definition of dimensions main grid and hidden grids with preloaded standard labware such as a waste container Deck templates can be customized 610766 02 Page 51 of 225 Microlab STAR User Man
177. olab STAR Option and assembly with which the work surface of the Microlab STAR is covered in such a way that it is shielded from user intervention and other outside influences such as dust At the same time it protects the user from the movements of the Microlab STAR It is transparent Deck The work surface of the Microlab STAR It presents at the same time the greatest possible area cf Work Area The placing of the carriers on it is defined by the 55 tracks as long as they are in the operating range of the pipetting area Dispense To distribute quantities of liquid from a pipetting device Firmware Programs Sequences of commands which are carried out on the processors of the Microlab STAR Hardware error Type of error which depends exclusively on faulty functioning of the hardware Instrument Hardware of the Microlab STAR mechanics electronics and firmware 610766 02 Page 211 of 225 Microlab STAR User Manual Instrument commands The commands made available by the firmware for controlling the Microlab STAR Labware Refers to movable items to be placed on the Microlab STAR deck such as carriers containers or racks LIMS Higher level data processing system generally Known as Laboratory Information Management System also LMS Liquid Includes all kinds of liquids among which are included reagents controls standards wash fluids LLD Liquid Level Detection Positive pipetting of liquid
178. olume in the first part of the dispense step e Blowout volume blow out air is taken up first during aspiration if dispensing will later be done using the empty tip dispense mode In the dispense step the entire volume including blow out air is dispensed Page 102 of 225 610766 02 Microlab STAR User Manual e Swap speed is the speed at which the dispensing head is drawn up out of the liquid e Settling time is the time the dispensing head has to wait in the liquid after aspiration dispensing until it begins to withdraw e QOver aspirate volume is a kind of pre wetting volume on aspirating e g 20ul of liquid first more than 20ul is aspirated 20uI Over Asp Vol so as to pre wet the tip Then this volume is immediately dispensed again still in the aspirate step e Clot retract height a parameter for recognizing blood clots which determines how high the dispensing head is allowed to travel up out of the liquid if there is a residual cLLD signal after aspiration It is measured in mm from the height of the liquid surface upwards If this distance is exceeded an error message is generated e Stop flow rate dispensing speed of the plunger expressed as a stream of liquid volume in ul s at which the dispensing step terminates abruptly If dispense flow rate is equal to stop flow rate the dispense breaks off abruptly after dispensing the volume without slowing down beforehand
179. on to select a library Use this dialog to import all the libraries you need and to remove libraries you no longer need This example links to the HSLML_STARLib hsl library This library contains some interesting functions for the Microlab STAR Note that if your method uses a library function the library in question is indicated in the list of referenced libraries and can t be removed You can also write your own library function using the HSL Method Editor see chapter 9 HSL Method Editor page 96 and use it like any of the standard libraries 610766 02 Page 91 of 225 Microlab STAR User Manual Once you close this dialog with OK the mathematical library functions for example are available under their own bar and can be invoked like standard commands In this example the following functions are linked and now made available within the method Pressure based aspiration monitoring off Pressure based aspiration monitoring on Capacitance based clot detection off Capacitance based clot detection on the clot heigth must be specified in the liquid class Measure container volume with n channels auto increment 1 yes O no if pattern is reduced e g 11000111 all sequence positions are used Get container volume of i channel in ml as measured in previous Measure Container Volume Step Set threshold for pressure based clot detection Additional functions available include the generation of a Microlab AT
180. on of the surface and then dispense by following the rising liquid level For volumes larger than 20 ul the liquid can be dispensed in a jet without touching the surface To dispense in a jet specify a position a couple of millimeters above the surface and dispense by following the rising liquid level For dispensing in a jet a varying amount of blowout air is used to make sure that all liquid is dispensed from the tip However dispensing to the surface of an empty vessel touch off is also possible For dispensing into an empty container position the tip less than 1 mm typically 0 4 mm above the bottom of the container and dispense following the rising liquid level After the dispense the situation within the tip looks like this Channel Blowout air optional Liquid Stop back volume ootional Transport air as specified Transport air as specified on dispense optional on aspiration optional Situation on dispense Firstly transport liquid and blowout volume are dispensed followed by an aspiration of stop back volume in the case of partial volume dispenses and transport air Page 16 of 225 610766 02 Microlab STAR User Manual 2 2 4 Tip Drop Off The final step is to eject the used tip into the waste container of the Microlab STAR A needle will be placed back into the wash station where the wash process may be started directly 2 3 Avoiding Contamination If cross contamination is a concern consider the
181. on the desktop Then the method or the deck layout of interest must be opened from the run control by selecting File gt Open from the menu On opening the method a dialog box appears for a period of time informing the user that the method is being parsed i e analyzed This procedure may take up to a minute depending on the size of the method Parsing in progress __ Method will be parsed please walt Parsing a method Page 98 of 225 610766 02 Microlab STAR User Manual After loading and parsing a method you can use the Start button to execute it i Hamilton Run Control TubesToPlatePipette lay File View Window Help 5 50 ae AS ral en 30 35 40 4 lee Tubes T oPlatePipette_lay fee TubesT oPlatePipette_lay 2122142 SYSTEM Parse method start Method file C ProgrammeSH4MiL TON SMethods De 21 24 58 SYSTEM Parse method complete Single Step Pause Abart 0 processed approximately For Help press F1 A method ready to run Each executed instrument step is logged in the log window The choices within Run Control are e To start a method e To abort a method e To use single steps system pausing after each single step e To pause a method to be continued later Clicking on the deck layout view of the Microlab STAR within Run Control adds the Tools menu to the menu list Here you will find shortcuts to access the configuration and liquid editor The configuration ed
182. ons and e decreasing y positions from channel 1 rear most to 16 front most To program a method using single steps e Select the option keep pattern within the Channel Settings dialog in all single steps tip pick up tip eject aspirate dispense Do not use the option all sequence positions e Even if an error occurs and Exclude Channel is selected as an error recovery the pattern will be kept To program a method using SMART Steps e Select Copy Pattern e Do not select Exclude Erroneous Positions e f sequences are reloadable in runtime during the pipette command deselect the option reducible by user to maintain the original block wise structure of the sequences 610766 02 Page 95 of 225 Microlab STAR User Manual 9 HSL Method Editor The HSL method editor gives the user access to the underlying HSL code offering higher flexibility for programming methods Usually this method editor is used only to customize methods written with the graphical method editor Of course a complete method can also be written using the HSL method editor The HSL Method Editor is a general purpose text like editor that can be used to create methods based on HSL HAMILTON Standard Language for any instrument system It is a sub component of the User Software Parameters for instrument steps are entered using dialog boxes and may include variables The editor makes use of the same instrument specific s
183. ontainer The teaching station with needle Right click on the labware to be taught within the deck layout Select Move Labware from the menu Click on the Move Probe button A needle is picked up from the teaching station Note that the teaching station does not have to be added to the deck layout software Move Probe Rey Control Page Up 4 1034 30 mm 0K L y 345 90 mm E i 2 50 00 mm We Step IT Advanced e a Probe Tip increase step size decrease step size Use arrow keys and page up down keys to move probe Use keys to ncreasedecrease step size respectively Page 56 of 225 610766 02 Microlab STAR User Manual The dialog gives the current x y z coordinates and allows you to move the needle to any position on the deck The Advanced section allows you to move directly to a position by clicking on Move Probe Move Probe Advanced Select probe Probe Tip 0 Cancel Key Control Move to position K 208 00 mm y 530 00 mirn Z 21 2 60 mm Move Probe Clicking on Key Control brings you back to the previous dialog Clicking OK ends the teaching process and writes the current position into the x y z coordinate fields of the move labware dialog Alternatively select Move Probe from the tools menu NOTE The reference points for teaching are the tip of the needle and the reference well of the rack which is usually mar
184. or Auxiliary Page 42 of 225 610766 02 Microlab STAR User Manual Alternatively the user may start with the method editor as the master editor if there is no shortcut to the method editor create it manually The relevant executable file is hamilton bin HxMethEd exe Labware Editor Auxiliary Microlab STAR Meth ML_STARSysiem ML_STAR Method Editor is Master Deck Editor is Slave Foe Help pres F1 NUM Sequence Editor Auxiliary delay ondi z 4 gt is 05 see Run Control is Slave The auxiliary modules and tools small arrows in the above figures are called from the tool menu by menu items or tool icons Finally the method is executed using the independent run control In some cases one may wish to define special labware items or sequences pipetting patterns see below using the labware editor and the sequence editor 4 2 Methods and Deck Layouts The Microlab STAR User Software allows an n n relation between deck layouts and methods This means that one deck layout may be used for different methods or one method uses different deck layouts e g to link two Microlab STARs or a Microlab STAR and a Microlab SWAP The run control allows you to start either deck layouts with the corresponding method if there is only one or in general methods These methods are then either executed or simulated 6107
185. osition probe at center of container at last position omea Rotate Position Fixed deck Free form Deck position coord mm Y coord mm Z coord mm First position fo fo fo Move Probe Position probe at center and bottom of container at first position Last position fo fo fo Move Probe Position probe at center of container at last position Use the Rotate button to rotate the labware item in increments of 90 degrees and type the position in X Y and Z coordinates into the input boxes under Deck Position Click OK If the labware item is to be placed at an angle other than 90 180 270 or 360 degrees the Free form mode will have to be used Here two sets of coordinates are required 610766 02 Page 55 of 225 Microlab STAR User Manual 6 2 3 Teaching Labware Teaching means guiding a channel or other probe to a particular location on the deck and assigning a name to that location The User Software enables you to record the x y z coordinates of that point and to associate them with the name you have given the location Afterwards you need only specify the name of the location and ML STAR remembers exactly where it is on the deck The precise position of labware items can be taught in this way using the needle from the teaching station This is an accessory The teaching station is placed on track 55 next to the waste c
186. ource_1 and Source_2 and two target plates here without name onto the deck Two tip rack are needed as well Save the deck layout under the name HitPick 610766 02 Page 143 of 225 Microlab STAR User Manual This method modifies a sequence the hit sequence on runtime The next step is therefore to generate a sequence hits using the sequence editor The sequence should span some or all of the wells in the source microplate s Start the sequence editor from the deck layout editor by selecting Sequence Editor from the Tools menu Zoom in to view the source plate Rubber band the entire plate so as to add positions to your new sequence PS Hamilon se Elon Bae Conia cence P e e ed amp Bs 61 Q W E fien BEX gt if Wasi o5 seconds r BE ee ee 6660066000000 2 tart 2o _ s 0606060606086 000000000000 6 Tast b 660606060006000606006080 000000000000 6060600606006000600000 Tarse ho 6 06600600606060680 000000000000 4 F of EAE Pick a corner Pick a direction ENE _ Serpentine alfa Te Sort Racks A Sortby Grid Columns A Sorthy Grid Rows A Fitter by Probe Head For Help press F1 Save this sequence under the name Hits Create another sequence spanning the two target plates and call it Target The last sequence to be prepared is the Tip sequence holding both tip racks Save the changes to the sequence edito
187. ow at the bottom shows the results of the syntax check if performed If any error is listed in this view double clicking on it will set the cursor to the incorrect statement in the Method View If the HSL method editor is used to edit a method that was written with the graphical method editor insert the HSL code only between two adjacent comment lines Leave the comment lines and all code in between unchanged Save your changes and close the HSL method editor If this method is reopened with the graphical method editor your changes will be visible as a greyed out unchangeable HSL code block 610766 02 Page 97 of 225 Microlab STAR User Manual 10 Run Control Run Control enables you to execute your methods During execution of a method the instrument steps are automatically logged and displayed in the log window as well as written in a trace file with the same name as the relevant layout file The Run Screen shows a static view of the deck layout Because there is an open n n relation between methods and deck layouts see chapter 4 the Run Control may start either deck layouts lay or methods hsl A further alternative is to start linked methods cmt The run control may be accessed by the shortcut buttons of deck layout or method editor SLEI Then the method of interest is automatically loaded into the Run Control Alternatively the Run Control may be started by double clicking the Microlab STAR Run shortcut
188. plate called Plate Save the deck layout under Aliquoten lay Open the method editor and select Instruments from the editor s Method menu to link the method to be written to the given deck layout Aliquoten lay as described in the previous examples Here s the method B Aliquoten med O x al Pit Instrument short name ML_STAR load 3 sequences ML_STAR Tips ML STAR Samples ML STAR Plate Moh Pipette with Tips from ML_STAR Smart Steps We te Instrument short name ML_STAR Aliquot procedure Volume per well pl 20 Aspirate sequence ML_STAR Samples Dispense sequence ML_STAR Plate controlling Sequence argo Unload from ML_STAR Smart Steps l Instrument short name ML_STAR unload s sequences ML_STAR Plate ML STAR Samples ML_STAR Tips Page 140 of 225 610766 02 Microlab STAR User Manual The loading and unloading is very similar to the previous examples and may be skipped see note in example OnePlateToPlatePipette Let s concentrate on the pipetting step Drag the SMART Step pipette to the method General Tip sequence Instrument short name ML STAR M eee rca a E ML_STAR Tips Standard pipette procedure f Aliquot procedure W Reducible by user Pipette mode f Simple Mi i Replica in t Pooling lnd Tip handling replacement Multiplicity n o After each dispense
189. position A3 A4 etc The desired multiple can be chosen e g 2 for a double test Pooling for cycles of multiple aspirations dispenses where liquid from multiple source containers is dispensed into one target container e g tube 1 tube 2 gt plate position A1 tube 3 tube 4 gt plate position A2 etc Here too the desired multiple has to be chosen e g 12 for pools of 12 For the aliquoting procedure the choices under pipetting mode are deactivated Aliquoting means that one aspiration is followed by multiple dispenses e g tube 1 gt plate positions A1 to A12 The next step is to select the appropriate tip handling Tip sequence ML_s TAR tips_ 31 M Reducible by user Tip handling replacement After each dispense One tip set After fanstered volume None i After each sample Tip counter Tip counter white tip counter First select the tip Sequence where the tips are to be picked up from The tip type is automatically retrieved from the sequence If the tip sequence is used up it will be automatically reloaded If Reducible by user is checked the tip sequence can be reduced on reloading Tip handling can be chosen to replace tips after each dispense if multiple dispenses are needed to transfer e g 900 ul with a standard tip of 300 ul Page 72 of 225 610766 02 Microlab STAR User Manual after each sample multiple dispenses of the same
190. program Variables can be used for the vacuum box number and duration of pumping action NOTE The capacity of the waste tray is 200ml Program your methods to activate the vacuum box waste pump before the capacity is reached so that this tray does not overflow 610766 02 Page 111 of 225 Microlab STAR User Manual Move command a Move Vacuum bos number Move to OK Cancel Help This command moves the carriage from one chamber to the other Select the chamber over which the carriage is to be moved The conditioning chamber is in front and the elution chamber in rear If the vacuum box is not yet initialized when it receives its first Move command it will initialize first and then move the carriage to the desired chamber If it is already at the position specified in the Move command the carriage will not move Vacuum command aR Vacuum Yacuum Parameters Vacuum box number fi Chamber AULT Inches of Mercury Ramp duration 20 seconds Yacuum duration E0 seconds Error Recovery OE Cancel Help This command runs the vacuum pump inside the vacuum box controller The vacuum box number and the chamber have to be selected Page 112 of 225 610766 02 Microlab STAR User Manual There are three vacuum parameters to be defined The vacuum in inches of mercury Hg 1 inch Hg 33 86 mbar is dependent upon your protocol and the limits set by the filter p
191. quence and the variable storing the name of the tip counter NOTE Either one of the two steps involving the tip counter may be sufficent The edit step also sets the counter to the position read If no request for a manual reset is needed the read step alone may be used Then sample reduction from the user input is done by setting the end position of the sequence AllTubes to the number as input BA Sequence Set End Position Edit Set end position of sequence to position ML _STAR AlTubes No5 amples Cancel Help The pipetting loop thus loops over the sequence AllTubes terate over sequences O oi ue O O ee e Omsan O O OOOO o i i id Consequently the adjust sequences step is used to handle sample reduction if less than the full number of channels are needed Page 162 of 225 610766 02 Microlab STAR User Manual Adjust Sequences Edit Sequences C eene O Consumed times 5 WL STAR AllTips ML_STAR AllTubes ka ML_STAR plate Add all the sequences shown in the dialog and recall the notes in the previous chapter For the pipetting single steps the settings are as in the other examples Tip Pick up Sequence AllTips counting automatic Aspiration Sequence AllTubes counting automatic Dispense Sequence plate counting automatic Then the current position of the tip sequence is written into the tip counte
192. r Write of HSLTipCountingLib Edit Library name C Programme HAMILTON Libray HSLTipCountingLib hel Variable Function name TipCount Awite P Function parameters Parameter Description Sequence lbh ML STAR AllTips tiplounter Tipcounter variable 610766 02 Page 163 of 225 Microlab STAR User Manual Now a further possibility will be demonstrated how to make decisions within the method Suppose a user output is programmed Kil User Output Edit Dialog title o utput Buttons Default button es and Ho buthon No Icons Display Information message icon Return value fo utputA eturr al Sound gt Timeout 3 M Infinite Output Text Unload the tip carrier The dialog at runtime prompts for the tip carrier to be unloaded and shows a YES and a NO button If the user clicks YES the return value of this dialog will be 6 if he clicks NO the return value will be 7 This value is stored to the variable OutputReturn as specified in the dialog The decision is made depending on the value of this variable The if then construct compares the variable to the fixed value 6 YES clicked Only then is the code between the if and the end if statement executed Lett operand Comparison operator Aight operand egual to B F Include else block This code is the unloading command for the tip carrier
193. r Tip Patent Pending PCT EP 00 03423 The Air Displacement Pipetting Principle The air displacement principle has the following advantages e Independent and modular design of pipetting heads for most flexible assay programming e CO RE compression induced O ring expansion technology for the flexible coupling of tips or needles to the pipetting channel e Tips or reusable needles of three sizes on the same head in the same run Page 12 of 225 610766 02 Microlab STAR User Manual e The construction principle enables pressure based liquid level detection and aspiration monitoring e No contamination or dilution by system liquids e No drops due to moving tubes e No problems with corroded tubing pumps etc e Same commonly accepted pipetting principle as for hand pipettes 2 2 From Aspiration to Dispensing In this section we describe in detail the processes involved in a simple pipetting step 2 2 1 Tip Pick up The first task for the Microlab STAR is to pick up a disposable tip or a reusable steel needle For tips special carriers typically holding 5 tip racks of 96 tips are placed on the instrument deck Steel needles may be picked up directly from the wash station or from a separate needle rack which typically is a tip rack on a tip rack carrier with needles instead of tips 2 2 2 Aspiration The first step within an aspiration and dispense cycle is to aspirate a variable amount of blowout air which is
194. r both Submerge Depth 2mm following liquid level Dispense Capacitance LLD on following liquid level Page 18 of 225 610766 02 Microlab STAR User Manual 3 Aspirate gt 20ul of a water like liquid dispense the same amount into an empty 96 well plate keep tips Liquid Class Aspiration Mode Dispense Mode Detection Comment StandardVolume_Water_DispenseJet Aspiration Jet Dispense Empty Tip empty tip only Aspiration Capacitance LLD submerge depth 2mm following liquid level Dispense Fixed height of 5mm not following liquid level On first aspiration pre wetting of the tip by 1 3 mixing cycles is necessary to equalize conditions for initial and subsequent dispenses 4 Aliquoting of liquid means aspirating a given volume all at once and dispensing several partial volumes aliquots in a jet to different containers In this frequently used pipetting procedure measurements have revealed that the accuracy of the first and the last aliquot are often not within the specified range Therefore to dispense e g 10 aliquots of 20 ul of a water like liquid with the ML STAR aspirate 240 ul and dispense 20 ul directly back into the container This is followed by dispensing 10 of the 20 ul aliquots The last aliquot of 20 ul is discarded into another container or ejected with the tip In addition after the dispense of every aliquot a given amount of air is aspirated and dispensed with the next aliquot Liquid Class Asp
195. r name and definition selected must always be identical to the name with which the physical carrier is labelled Here are some frequently used Hamilton standard carriers Plate carriers for 5 flat 96 or 384 well microplates PLT CAR L5MD Plate carriers for 4 1536 well microplates PLT CAR L4HD Plate carriers for 5 96 deepwell or archive microplates PLT CAR L5AC Tip carriers for 5 tip racks of 96 tips each TIP CAR 480 Preloaded carriers for 32 or 24 tubes with defined diameter and length CAR24 Cup15x75 Plate carriers for 5 96 well PCR microplates PLT CAR L5PCR Plate carriers for 3 96 or 384 well flat microplates in portrait orientation PLT CAR L38MD Plate carriers for 3 96 deepwell microplates in portrait orientation PLT CAR L3AC Carrier for 12 reagent containers of 100 ml each REA CAR L3AT Temperature controlled carrier No 1 and No 2 CAR_TCC_1 CAR_TCC 2 Wash Station No 1 and No 2 for needles CAR Wash 1 LowNeedle etc Page 52 of 225 610766 02 Microlab STAR User Manual Carrier definitions are stored under hamilton labware ML_ star Files defining carrier templates have the extension tml A carrier features a varying number of locations for the placement of labware such as tip racks microplates etc To add a carrier to the deck layout simply double click on the appropriate track position The add labware dialog box opens up Type in a name for the carrier and click browse to select the PLT CAR_L5MD from the labware ML
196. r on quitting Click the Edit Method Button From the editor s Method menu select Instruments to link the method to be written to the given deck layout HitPick lay as described in the previous examples Page 144 of 225 610766 02 Microlab STAR User Manual This method uses library functions To link the libraries to your method select Method and Libraries from the menu Click on the browse button and link the two libraries HSLTrcLib and HSLSeqLib to the method Click OK Libraries Add Library Remove Library Cancel Help The functions available within these libraries are used to generate entries into the methods trace file whenever a Hit is found and to manipulate sequences in the appropriate way 610766 02 Page 145 of 225 Microlab STAR User Manual The items of this method in the graphical Method Editor are displayed on the next screen User Input Dialog Title User Input Return Value Sound Timeout infinite Inpe threshold Enter the threshold for the optical density String 1 0 Assignment with Calculation eqlselect SELECT FROM absorbance WHERE OD threshold Comment Remove all positions from sequence Hits seqrkemaveAll af HSLSeqLib seqrkemovesli ML_ STAR Hits File Open File handle filet File name AdsDatas xle Table name Absorbance Mode Open file to react Columns la
197. re defined to determine handling of the labware elements These properties should not be changed by the user They are accessible just so that you can have the maximum of decision making power in particular situations that call for it These properties can be divided up into the following groups e Information for the handling of the auto load unit e Information for the special units like wash carrier TTC tips and needle handling waste etc e Information to support the reduction of selection during edit time 17 4 1 Structure The properties are always combine a key name and a value The key names are case sensitive the distinction between capital and lower case is important All values are in integers no decimal points 17 4 2 Information for Handling the Autoload Unit MlStarCarWidthAsT 1 n Width of carrier in T MiStarCarCountOfBCPos 0 n Count of carrier barcode position How many barcodes positions are pd pected S O MIStarCarRasterWidth 0 1 mm Distance middle of barcode position to middle of next barcode position MiStarCarBCOrientation 0 0 or 1 0 Vertical 1 Horizontal barcode read direction MiStarCarFirstBCPos 0 1mm Distance between rear of carrier to middle of first barcode position MiStarCarBCReadWidath 0 1 mm Width of barcode read window MiStarCarlsRecognizable 0 Oor 1 The carrier has a magnetic bar so its presence can be detected 1
198. rn string black separator The step selection finally allows you to exclude steps such as the move commands from the method editor Page 48 of 225 610766 02 Microlab STAR User Manual Only if instrument is selected within the configuration editor is the additional tab communication settings visible Communication settings Instrument configuration Communication settings f R5232 Com port com f USB Select the type of communication and the com port for communication via RS232 interface Click save to activate the new settings or abort to leave the settings unchanged 610766 02 Page 49 of 225 Microlab STAR User Manual 6 Defining the Deck Layout In the Hamilton Deck Layout Editor the user is prompted to tell the system which labware carriers racks or containers is to be used in the procedure and where these items are placed on the deck This is called defining a deck layout The Hamilton Deck Layout Editor is started by double clicking the shortcut icon displayed on the desktop of your PC iI Untitled Hamilton Deck Layout Editor File Edit View Tools Help Ojea Le AIS WPI Run Method For Help press F1 Deck layout editor The method editor and the run screen can be started from the deck layout editor by clicking the appropriate buttons located in the leftmost frame For access to the main modules a toolbar is provided In that toolbar the following ch
199. rrier Temperature in CO Float 10 0 fe SetCarrier Temperature on ML_STAR ir Rack type TempCarrier Temperature 1 Ambient C Return value count 3 Edit of HSLTipCountingLib TipCount EdittMl ST amp R AITips TipCounter variable ML_STAR 7 Wj Sequence Set End Position aL end position of sequence ML_ gt TAR AllTubes Mosamples Comment lt Start Pipetting Sa over folowing Sequences ML_STAR AllTubes oop Countert used as loop counter variable a J tn tal he Ln J a co Page 158 of 225 610766 02 Microlab STAR User Manual Adjust Sequences Adjust the following sequences ML_STAR AlTips C1 times consumed ML_STAR Allfubes 1 times consumed ML_STAR plate 1 times consumed TipPickUp on ML_STAR T Return value count 3 Aspirate on ML_STAR settings on Capactive 4 Return value count 3 Dispense on ML_STAR settings off Liquid following an Return value count 3 TipEject on ML_STAR 20 Return value count 3 21 End Loop ML_STAR AllTubes Reset Sequence after loop 22 Write of HSLTiptountingLib Tiplount Vvrite ML_STAR AlTips TiplounterYariable 23 Comment sUnload Tip Carrier on reguest 24 User Output Sound Timeout infinite Cutput Unload the tip carrier 25 If OutputReturn is equal ta 6 26 UnloadCarrier on ML_STAR Wil Rack type tipcarrier Return valu
200. ry key HKEY_LOCAL_MACHINE SOFTWARE Phoenix instruments MicrolabS TAR Method Editor has to be changed from HxMetEd to HxHSLMetEd We recommend that you create a shortcut on the desktop to the HSL Method Editor The editor is stored under namilton bin HxXHSLMetEd exe 18 1 Create a Method to Copy from Plate to Plate The method we are going to describe copies wells i e aspirates liquids from wells on a plate and dispenses them to the corresponding wells on another plate A1 A1 H12 5H12 The transferred volume will be 50 ul The method name is HSL_OnePlateT oPlate First an appropriate deck layout has to be created and saved as HSL_OnePlateToPlate lay The deck layout for this method is shown in the screenshot 16 Hee ae Pin Decklayout HSL_OnePlateToPlate To create this deck layout start the Deck Layout Editor by double clicking on the appropriate icon Select Open from the File menu then select the deck layout from the example in chapter 13 3 OnePlateToPlate lay Save this layout under the name HSL_OnePlateToPlate lay Alternatively create the new deck layout again by adding labware Page 204 of 225 610766 02 Microlab STAR User Manual Open up the HSL Method Editor and load the HSL_OnePlateToPlate HSL file Under the View menu on the menu bar check View All Stepdata so that all the parameters we enter will be displayed on the screen EJ Hamilton HSL Method Editor HSL_O
201. s of a loading head movable in Y direction which draws the carriers into the Microlab STAR and can read the barcodes on them AutoLoad tray Hardware unit On it the carriers can be placed and held outside the Microlab STAR The loading tray is attached to the Microlab STAR to support the automatic loading and unloading process Barcode Mask The barcode mask defines the basic structure of a barcode It is a pattern to which a barcode must conform The assignment of a specific labware item can be done this way The barcode mask can require a barcode to contain specific strings at fixed positions It can contain wildcards too Barcode Reader Part of the Autoload Option Basic Microlab STAR Basic parts of the Microlab STAR with pipetting arm and deck to which the loading unit and the options can be added on Page 210 of 225 610766 02 Microlab STAR User Manual Carrier Manipulable unit for loading on the Microlab STAR Hardware unit in or on which elements stand and which is transported when the Microlab STAR is loaded cf Autoload Container Includes test tubes reagent containers and wells Container identification Barcode for identification of containers Serves for unambiguous identification of the vessel e g a sample test tube Continuous loading Refers to the loading of manipulable elements onto the Microlab STAR after processing has been started Front Window and Side Cover Covering for the Micr
202. s system simplifies variable usage by allowing implicit declarations and by supplying complete variable context to edit dialogs The output of the Method Editor is a linked set of files that may be executed by the Run Screen or further customized using the HSL Method Editor see chapter 9 HSL Method Editor page 96 In most cases but not necessarily a method references one single deck layout The Method Editor is accessed by clicking on the Edit Method button in the Deck Layout Editor after the deck layout has been defined and saved e g as MyDemo lay Alternatively the method editor may be started directly from the desktop 3 HxMetEd MyDemo med Oy x File Edit View Method Tools Window Help Osua sm x dh cot a le General Method Editor The Method Editor is divided into 2 sections The Method View on the right is a grid containing the method The Toolbox Window on the left offers various elements that can be used to build the method grouped by bars Page 66 of 225 610766 02 Microlab STAR User Manual At the beginning of a new method there is only one bar named General containing the common language elements such as if else loop function call etc No instrument specific steps e g aspirate are visible so far All steps can be disabled enabled on request by right clicking a step or a block of steps and selecting disable enable from the menu It is possible to load different methods in
203. sccssccsces 202 18 Reference Guide Examples Using the HSL Method Editor ccccccsssssscccccssssssccccees 204 18 1 Create a Method to Copy from Plate to Plate cccccccssssssssssccccccccssssssssccccsseeeees 204 APPENUICOS esse e E sete dah ul O cb lea enous its E sauvecbaatucleas 210 Pic GIOSSATY srciteieicalaehedeaesieeaiaictacetawsliccdaastened eaaaciisatuendexcsnnatiseoweaedezsneucisasuebieassassniseudeeedessuesssnes 210 B AMOK ciiscssccecerecesscevepcsscsceasccesnaesscierestavausesesacsuanavacanautseee ceaeovausansescswensccestepteaeavaabsevsasasessessasccsasnes 216 Co MREGUTALONY A HAYS isien oasian aa o eaer aa a an aiei 219 Ds Ordering Tntor mao ns ccisenscs cossincedataswctecessccten saar eae a aa aeai a aiaa S 221 Ks Principles of Cali Prato sisses aaaea eaa aa aa aa aiana anaiari 222 F Needle Wash Station Chemical Compatibility eeeeeooossssseeccocsssssseceoosssseseceoosssssseeesosssoe 223 Page 6 of 225 610766 02 Microlab STAR User Manual 1 General Information Hamilton s Microlab STAR is the next generation pipetting workstation This User Manual is designed to help you get the most out of your Microlab STAR You should read through the entire manual before beginning to operate your instrument This first chapter should be read with particular attention It contains important information about the use of the Microlab STAR and this manual 1 1 About this Manual This manual is to help users
204. se Offset Visible Draw Click on the Add button and double click the added line imenszions Origin a mm SITE T length q lt gt width Origin r 236 5 mim Origin Z 177 6 mm Width 12 mm Length o6 mm Drawing M Draw label M Draw site boundary Origin y Snap to site C Locate base of rack on template site Template Cancel The drawing on the right illustrates the different coordinates The values refer to a standard microplate Drawing is only of optical influence accept the defaults For Snap to site select the lower option for a microplate because a microplate fits with its bottom on to the plate carrier to enable a good electrical coupling for capacitance based LLD The decision whether a plate or tip rack fits on to a site of a carrier is made depending on the width and legnth of the site all plates that have the same boundary measures width and length can be placed on the site Different plate types 96 flat and deep well 1536 well etc have the same boundary measures 610766 02 Page 195 of 225 Microlab STAR User Manual You may add additional sites to the same carrier Click next Now the site can be preloaded Template Site Labware E4 Labware Files Sites and Labware nunc sun SB FL Lrek nuncu FB FL Irek Click on Browse to browse for the corresponding labware and click Add to add the selected labware to the site Click Finish Save the n
205. sh_1_standardneedle tml from the ML_Star directory Save the deck layout under the name TubesToPlateWithNeedles lay Create a sequence holding all tubes called Samples and a second sequence holding all wash modules and name it WashStations using the sequence editor as described in the previous examples Click on Edit Method and link the deck layout to the method to be written 610766 02 Page 153 of 225 Microlab STAR User Manual Here s what the method finally looks like Load from ML_STAR Smart Steps l Instrument short name ML_STAR load 2 sequencers ML_STAR Plate ML_STAR Samples Comment lt Pipette tubes to plate and wash needles Needle Wash Settings from ML_STAR Smart Steps Instrument short name ML_STAR Wash sequence ML_STAR Yvashstations Start with wash solution one Wash solution ane Rinse time a s Soak time s 1 Stream intensity 3 50 Wash solution toy Rinse time a 0 Soak time 0 Stream intensity 0 Drying time 30 Number of air pulses 30 Air pulse duration a 0 3 Time to first air pulse s 1 Time between air pulses a 0 1 over following Sequences ML_STAR Samples oopCountert used as loop counter variable Adjust Sequences Adjust the folowing sequences ML_STAR Plate 1 times consumed ML_STAR Samples 1 times consumed ML_STAR VWashStations 1 times consumed Needle Pick Up
206. sion proof When working with flammable or explosive fluids or vapors the necessary precautions must be taken 3 3 4 Temperature Controlled Carrier TCC The temperature controlled carrier is a device to heat and cool microplates The TCC has four positions for microplates which are all of the same temperature The TCC may heat microplates up to 60 C or cool them down to 22 C below ambient temperature The TCC is able to read barcodes on the microplates with the autoload option It consist of a plate carrier with peltier elements on the instrument deck and a service station with a heat exchanger below the bench For the heat exchanger a water based heat exchange liquid is used in a closed cycle it is stored in a reservoir Typical times at 40 rel humidity to heat and cool the TCC are Tambient 20 C Tambient to 60 C 20 mins 60 C to Tambient 20 mins Tambient to 4 C 15 mins 4 C to 60 C 25 mins The temperature controlled carrier TCC 610766 02 Page 31 of 225 Microlab STAR User Manual The heat exchanger solution reservoir NOTE Ensure there is always enough liquid 1 L within the reservoir Allow air exchange between the exchanger and ambient air The default position of the TCC is on the instrument deck Never leave the TCC unloaded on the autoload tray Replace the heat exchanger solution twice yearly Do not use SMART Steps to load a TCC with the autoload function Always ensure that the TCC is
207. start 13 10 03 Microlab STAR UnloadCarrier complete gt plt_ car l5md_ O00 13 10 03 SYSTEM Unload complete Hause 1310 03 SYSTEM Execute method complete Abon 100 processed approximately The method trace file is stored under OnePlateToPlate trc within the methods directory Each method trace contains the date the method name and an index within the file name a OnePlateToPlatePipette 20011212_1 tre The method traces are not overwritten or appended NOTE From time to time all unused method traces and com traces have to be deleted from the hard disk You may test your method using single steps It is always possible to execute only the next single instrument step like TipPickUp Aspirate Dispense etc by using the Single Step button After each step the system will be paused and the pause screen appears A method can also be paused clicking on the Pause button of Run Control Hamilton Hun Control OnePlateT oPlatePipette lay x Execution paused Currently no steps being performed Tou can continue the execution or abort the running method Single Step Abort Help Paused methods can be resumed and finished click on the pause dialog to stop the beeping Page 174 of 225 610766 02 Microlab STAR User Manual It is now possible during the pause to open the front cover of the Microlab STAR Before continuing the method make sure the cover is closed aga
208. t You must not end the worksheet name with a dollar sign Specify the sheet as sheetname where sheetname is Append to an existing the name of the worksheet worksheet in a workbook file Important You must follow the worksheet name with a dollar sign Specify the sheet as sheetname where sheetname is the Append to a not yet name of the worksheet existing worksheet in 4 workbook file Important You must not follow the worksheet name with a dollar sign NOTE You cannot specify a value in a range that exceeds the maximum number of rows columns or sheets for the worksheet or workbook For more information on these values see your Microsoft Excel documentation The format of all entries within one column of the excel file must be identical text number etc Page 94 of 225 610766 02 Microlab STAR User Manual 8 9 Writing Batch type Methods for a 16 Channel Microlab STAR As discussed in chapter 3 2 1 the 16 channel Microlab STAR is intended to be used as a batch like processor This means all 16 channels should aspirate and dispense simultaneously to allow maximum parallelization and highest pipetting speed In this case the reduced random access space of a 16 channel Microlab STAR does not cause any problems To achieve a batch like process all sequences involved must consist of blocks of at least 8 or better 16 positions Sequence positions within a block must have e the same x positi
209. tation 10 11 20 29 34 36 38 40 55 59 64 81 84 94 97 124 130 140 147 Well 8 18 47 55 59 61 65 67 89 90 109 131 142 155 157 159 169 178 187 192 197 216 218 Worklist 5 103 104 610766 02 Microlab STAR User Manual C Regulatory Affairs CE CSA and UL conformity are maintained for Microlab STAR See the Declaration of Conformity for the instrument reproduced on the next page Radio Interference USA and Canada This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to both Part 15 of the FCC Rules and the radio interference regulations of the Canadian Department of Communications These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment This equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the present user manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause harmful interference in which case the user will be required to correct the interference at his own expense Pursuant to the Canadian Radio Interference Regulations ICES 001 Notice for Industrial scientific and Medical Radio Frequency Generators this ISM apparatus meets all requirements of the Canadian Interference Causing Equipment Regulations Please note
210. teps as the graphical method editor It allows methods to be created saved modified and printed To use the HSL Method Editor e Open Windows Explorer e Go to folder Program files Hamilton Bin e Create a shortcut to the executable file named HxHSLMetEd exe on the desktop Use the HSL Method Editor shortcut you have placed on the desktop to start it Then use the New menu to create a new method based on an existing deck layout or open a previously written method by means of the File gt Open menu Ela HSL Method Editor Demo hsl _ Oy x File Edit View Insert Tools Window Help e x JO eh S tee lor M a 4A44K Exe f aa or ee ee Do not insert text before this line 7 Aspirate Calibrate 1536Plate demoMethod Dispense FirmwareCommand Initialize LoadCarrier Load a carrier 7 MoveloPosition nl_ostar LoadCarrier 4 Initialize ML_STAR instrument 7 mlostar Initializef 3 Pick up tips mlostar TipPickUpe 3 UnloadCarrier spirate and dispense once with all channels til star Aspiratel tl star Dispensef 3 Eject used tips mlostar TipEject 3 library functions od Lb 4 Syntax Check HSL Method Editor Page 96 of 225 610766 02 Microlab STAR User Manual The HSL Method Editor window is divided into 3 sections The Method View on the right is a general purpose text like editor window containing the method written in HS
211. ter the life cycle of the instrument has terminated the Microlab STAR may be shipped to the original manufacturer or retailer Otherwise local disposal regulations are to be observed 3 10 Training Training in operation of the Microlab STAR and the User Software will be provided by HAMILTON personnel upon initial set up 610766 02 Page 39 of 225 Microlab STAR User Manual 3 11 Technical Specifications 3 11 1 Accuracy Trueness and Precision Specifications The pipetting specifications for the Microlab STAR are given in the following table The differences in the specifications are due to the accuracy of the gravimetrical measurement which to a great extent depends on the quality of the balance as well as on the stability of the environmental conditions pressure humidity temperature 300 ul Low Volume Pipetting Channel pecs Veil Vnpful IRU CVI IRI CV IR CV 200 lso h o Sts it so lao q do laes hW ee ao odao o ds dae de las bo bo yY 5 ao jas lhs psp e b5 Y a o 5 be Je 4 e dae 1 10 50 e pecs Veip ul Vipul IR CVI IR CVI IR CVI 1000 1000 p 0 75 1 5 1 2 0 1 5 500 1000 15 0 75 2 p 200 300 p 0 75 15 p 100 300 1 5 0 75 2 1 50 300 2 0 75 25 p 10 300 5 2 6 25 9 6 Vpip and V7 are the volumes of liquid and tip respectively Design specs are the design specifications which have been verified in the design phase of the Microlab STAR Final testing specificatio
212. that employed when adding a single labware position The block of positions is ordered according to default sequence for all labware object s involved 3 Control left click on labware position toggles the position s status in the sequence That means it inserts the position in the sequence if it s not already there same as left click on Page 60 of 225 610766 02 Microlab STAR User Manual laoware position otherwise it deletes the position with the largest sequence number from the sequence 4 Shift left click on labware position adds a block of positions to the sequence That means it adds all positions in the rectangle whose first corner has the coordinates of the most recently added position and whose opposite corner has the coordinates of the newly selected labware position The block of positions replaces the most recently added position and is ordered according to default sequence for all labware object s involved NOTE To select a labware position click on it only once If you click a second time on a labware position it will be added to the sequence again 7 1 2 Play Play first deselects all positions in the sequence then plays the sequence As each position is selected it is painted in the deck layout view and highlighted in the grid and tree views Play can be paused at any time and resumed by clicking on play again 7 1 3 Sorting Racks The Sort Racks dialog allows you to apply an order to labware positio
213. tically For example f 5 will generate a variable f of type integer NOTE To define a string text variable by assignment the text must be placed within quotation marks e g s Test s is then of type string Variables once defined show up in the drop down fields of all input fields of a method 12 predefined array variables of dynamic length are available in the array library For variable names do not e place numbers at the beginning e use blanks or signs other than the underscore _ Variable names are case sensitive To define a path within a string variable use the double backslash rather than the single backslash e g c program files hamilton methods For calculations with variables note that a 4 b 3 are integers c a b is therefore 1 also an integer To get the floating point result define a or b explicitly as a float by assigning a 4 0 an or b 3 0 Page 90 of 225 610766 02 Microlab STAR User Manual 8 6 Using Library Functions the ML STAR Library In addition to the toolbox elements several low level standard library functions are available such as the mathematical function library To use such functions you have to import the appropriate library Open the Libraries dialog menu Methods Libraries Libraries Doray Fie Namnespace HSLML_STARLikb hzl Add Library Remove Library i Cancel Help Libraries dialog Click the browse butt
214. tion is placed below the bench on which the Microlab STAR rests HAMILTON steel needles and disposable tips for the ML STAR 3 3 9 Needle Wash Station Page 28 of 225 610766 02 Microlab STAR User Manual The principle of the wash station is illustrated in the following figure where a typical procedure is shown Soak Drain STAR Fipauop Had STAR Fipavo Had Schematic drawing of needle wash process Needles are placed into the wash module washed from inside and along the outside with 1 wash and 2 rinse solution The pipetting channels blow air through the needles to expel any residual liquid from them One wash station consists of three individual 8 fold wash modules for the same needle type High Volume 1000 ul Standard Volume 300 ul or Low Volume 50 ul To enable parallel pipetting with an 8 channel Microlab STAR the following cycle of steps takes place 1 fresh needles are picked up from the first module for pipetting used and then placed back into the same module Washing starts 2 fresh needles are picked up from the second module for pipetting used and then placed back into the same module Washing starts 3 fresh needles are picked up from the third module for pipetting used and then placed back into the same module Washing starts 4 The next set of needles is picked up from the first module again which in the meantime has washed and dried all 8 needles The process is repeated again
215. to the editor and copy steps between the methods 8 1 General Method Commands A detailed description of all commands in the general method editor is directly available from the software Invoke the Help menu for more information A number of standard commands are available as you can see from the figure below For an explanation of the command buttons on each line numbered for convenience see the listing following the figure Whatever input is necessary for the individual commands all input fields labelled with text in brackets description contain optional information or the default may safely be accepted zl Z xl General ba General Comment Assignment m Assignment with Calculation 3 E Loop Break 4 T S Handling 5 fed File Handling E Timer Commands 6 7 Dialogs 7 1 Communication Commands E Shel B Set Event a E walt for Event 9 B Error Handling by the User H E Farallel Frocess Handling iw nB ML_STAR 14 ek P ML STAR Smart Steps General commands listing in icon and tree view Right click in the tool menu to switch between the different views 610766 02 Page 67 of 225 Microlab STAR User Manual Commands on line 1 Comment For any user defined comments in a method Assignment assign a variable Assignment with calculation perform calculations Commands on line 2 Loop Loop commands between start and end of loop Loop break T
216. tomatically closes sealing the container To reattach the container simply slot it back into its original position Wash Solution One red 2 right most containers 2 x 3 L Wash Solution Two blue left hand container 1 x 3 L The liquid containers of the wash station The waste container is placed to the left of the blue container and has a volume of 12 L Customers can order one of two complete variants of the wash station regular suitable for most uses and chemical resistant specially designed to meet the needs of those working with aggressive chemicals Each variant is complete with 3 wash modules and pump station For each variant there are high standard and low volume options The part numbers for the various options are given in the table below Regular OOOO O O Chemical Resistant LowVol hess stro O O oOo Sta Vo 0 182574 tore Highvo 182578 ts2720 O NOTE A table of chemical compatibilities with the wash station can be found in appendix F The information listed is based on laboratory tests with raw materials and should be interpreted as a guideline only Page 30 of 225 610766 02 Microlab STAR User Manual Chemical resistance is not the only criterion for the selection of wash liquids Also consider local regulations for handling and storage of wash liquids as regards toxicity contamination fire protection etc AN ATTENTION The needle wash station for Microlab STAR is not explo
217. tor window you can put the newly designed rack on the deck under Type select MyRack rck and the new rack is visible in the add Labware Window 17 3 3 Defining a Carrier Template Carriers are also pieces of labware Carriers have sites to host racks such as microplates Let us now define a carrier preloaded with flat 96 well nunc microplates NOTE A tube carrier in the sense of labware is a rack and not a carrier a template It is a rack that directly fits the track geometry of the microlab STAR and therefore can be directly loaded on to the instrument deck 610766 02 Page 193 of 225 To define a carrier open the labware editor and select New gt Template A window pops up Template Definition width 136 am Length 437 mm Clearance height 130 mm W Pre load sites with labware Background color Abbrechen Hilfe The width of the carrier is given by width mm number of tracks x 22 5 mm track In the case of a plate carrier which is 6 tracks wide the result is 136mm The length of a Microlab STAR track is 497 mm The clearance height for all carriers on the instrument is 136 mm Select the Preloaded check box to let the carrier be preloaded with the microplates Select a colour of your choice Click Continue Page 194 of 225 610766 02 Microlab STAR User Manual Within the next window the sites hosting the plates have to be defined Template Sites E4 Ba
218. ual 6 1 1 Save Deck Layout The complete loading of the deck may be stored under a chosen file name extension lay e g MyDemo lay This is done using the Save Save As commands in the File menu After a deck layout has been defined and saved you are ready to start programming your method 6 1 2 Open Existing Deck Layouts It is possible to load previously defined deck layouts extension lay by selecting Open under the File menu 6 2 Adding Labware to the Deck Layout Basically the deck is freely configurable The term labware refers here to carriers available from Hamilton as well as microplates from various manufacturers and tip racks The general procedure is first to add a carrier for a special kind of plate to the deck and then add the plate to the carrier There is a restriction however that carriers for one type of microplate only are available as standard parts Therefore do not place a deep well microplate and a flat microplate for example on the same set of vertical positions in the deck layout A ATTENTION If a tube carrier is placed directly adjacent to a plate or tip carrier using low volume tips may lead to a collision when the channel moves down to aspirate from the tube bottom 6 2 1 Adding Labware to Track Positions on the Deck The software comes with a set of ready made labware definitions for the standard labware items that are most commonly used in laboratories The carrie
219. ueous solutions only 610766 02 Page 21 of 225 Microlab STAR User Manual 2 5 2 Capacitance Based Clot Detection In addition to pressure based clot detection the ML STAR is equipped with capacitance based clot detection too This detection approach works for an aspiration with capacitance liquid level detection switched on The system measures the conductive signal when the tip leaves the liquid after aspiration Due to the air gap between tip and liquid the capacitance signal will vanish once a given height is reached the clot retract height which is specified within the liquid class If a clot is present it bridges the distance and the signal will remain resulting in an error message A typical clot retract height is 2 5 mm This is illustrated below Clot Retract Height This clot detection is independent of pressure based monitoring Page 22 of 225 610766 02 Microlab STAR User Manual 3 Description of the Microlab STAR 3 1 The Standard Microlab STAR STAR stands for Sequential Transfer and Aliquoting Robot The Microlab STAR performs pipetting operations on liquids in containers placed on its work surface The basic model Microlab STAR contains a work surface called a deck for placing movable carriers These carriers hold reagent containers such as tubes microtiter plates or other kinds of labware Front Shield and Window Pipetting Arm ee HAMILTON Micvolab STAR Ai N
220. used at the end of the last dispense to blow the liquid out of the tip This is done with the tips still in the air To start the aspiration of liquid the tip must make contact with the liquid This may be done by moving the tip to a fixed height This height must be chosen to be permanently below the liquid level to prevent the aspiration of air On aspiration the tip follows the falling liquid level if So specified according to the volume aspirated The distance to follow is computed from the known geometry of the first segment of the liquid container More elegantly and with greater safety the liquid level of the vessel to be aspirated from can be detected This can be provided by STAR s Liquid Level Detection LLD feature based on either capacitive cLLD or pressure pLLD signal detection For conductive liquids capacitive LLD should normally be used The sensitivity of the capacitive LLD that is to be used depends on the vessel size and the conductivity or polarity of the liquid that is to be detected For a solution of 0 1 NaCl in distilled water the sensitivities are 1 Very High 384 well plates 2 High 96 well round bottom plates 3 Medium 96 well flat bottom plates 4 Low Tubes 610766 02 Page 13 of 225 Microlab STAR User Manual NOTE Using an ionic buffer in your assay in place of distilled water may help to overcome liquid level detection problems Use only original Hamilton labware carriers For a proper c
221. usuenenstadassacwouehbadereensacuaueyeneas 28 3 3 4 Temperature Controlled Carrier TOC cccicieiieeiaiion ie e ee 31 610766 02 Page 3 of 225 Microlab STAR User Manual 3 3 The Automated Vacuums y stem AV 9 rena iageeid utente E OR 32 0 0 ANAP ener COURT T ETE OM PETTY Ee ewe TOTES TORENT are E eer Cran ie wea eeee 35 34 Computer Requirement Sessur niie a a aa a EEEa 36 Beall OON ihe ier E an aeranocecanetantacien sac eat an an lan ance getantacieenatPaeey 36 A SMI WANG A ANA A A EAEE ent eects ne sec EE AEE EA sec EEE TAA A EE 36 3S installation and SeU Piisi E E E EAE 36 DO a E VONAGE T E E EEEE A E E EEEE E S 37 30 Base Merola STAR persiennen a e E ier E NNE 37 302 Neede Wi KStars r a S Na 38 DT MD ENAN sean ea a EA Eaa 39 SiS WETMICALIOI Sis veestswadacecassesedesccsacsta sete sciwadeten o E A E O E E E T E A E S 39 3 9 Disposa lisons E AE EEAS 39 SU Trainine sa a a a a a a E e a EE 39 Ssh PeehlniCal S PeCmiCawOns scat cccesssesctiescs ec scssiselassiaueussaasehiesccenwsecassaduaseiawesacashcgusteusnesacesaseoatenes 40 3 11 1 Accuracy Trueness and Precision Specifications cccccccccsssssssssseeeeeceeeceeeceeeaaasseessseseeeeees 40 4 Desin OF the User SOfIWONC sirare e EA EEA E a 42 Bal CIV CGV IO iea a A E E 42 42 Methods and Deck Layouts sciis rennais anetares eaaa a aaia 43 4 2 1 Save As or Saving Methods and Deck Layouts under Different Names ssnssssssesosseeresessess 44
222. vered in this picture by the carriage A lid gasket in the carriage provides the sealing surface for the filter plate and a chamber gasket provides the sealing surface for the carriage In the picture on the right the carriage is on the way to the elution chamber NIDITIC NING CONDITION LT N HANIM BOX ELUTION The picture above shows the ports on the vacuum box There are three ports into which is fed the tubing that goes to the vacuum box controller There is also a push button switch that allows the user to manually operate the carriage An electronic control port is used for power and communication 610766 02 Page 33 of 225 Microlab STAR User Manual Vacuum Box Controller Vacuum Box 2 Vacuum Box 1 One Controller can control up to two vacuum boxes The front of the controller has eight ports Six of these are needed for conditioning elution and waste removal There is also a vacuum exhaust port for venting vapors and a waste out port for draining liquid from the conditioning chamber 24 Volt Power Connection USB Connection to the Connection to the Vacuum Box 1 Computer Connection to the Waste Box Pictured above is the right side of the controller with the different connections The communication with the computer is via a USB cable As the controller requires 24 volts a power supply with 24 volts is delivered together with the controller Page 34 of 225 610766 02 Microlab STA
223. volumeFilker _alycerin_DispenseSurface Glycerin Standard volume tip w StandardMNeedle water _DispenseJet water Standard volume neec High olumeFilter_Serum_DispenseSurface Serum High volume tip with Fil HighVolurnie Waker Dispensejet Waker High volume tip witho HighVolurme DMSO Dispensejet DMSO High volume tip witho Standardvolume Water DispenseSurface Waker Standard volume tip w Standardvolume DMSO DispenseSurface DMSO Standard volume tip w HighNeedle Water DispenseSurface Waker High volume needle StandardyvolumeFilter water AliquotJet Water For aliquot Standard volume tip w StandardvolumeFilker EbOH Dispensejet EtOH Standard volume tip w StandardNeedle w ater300ToSysFl_DispenseSurface Wakters00 to SvsFivater Standard volume neec LowNeede Water DispenseSurface Waker Low volume needle High olumeFilter _Slycering0 _DispenseSurface alyceringi High volume tip with Fil Standardvolume Water DispenseJet Waker Standard volume tip w HighVolurmneFilter DMSO DispenseSurface DMSO High volume tip with Fil StandardvolumeFilker DMSO Dispensejet DMSO Standard volume tip w Low Volume Glycerin Dispense Surface Glycerin Low volume tip withou HighVolume EFOH DispenseSurface EtOH High volume tip witho Standardvolume Serum DisoenseSurface Serum Standard volume ou 4 b F i i l d IJ JAA AAA A A A A A AA A A O O O O Liquid Editor Start Window 610766 02 Page 101 of 225 Microlab STAR User Manual Select for example StandardVolume_Water_DispenseJet Click E
224. w Sequence Bly Heb SR ERAN e AR REX r i Efese El zi a rawar _ _costar_flat_H 6 costar flat _6_costar_flat_l 6 costar flat_l1 6 _6_costar_flat_lt 6 costar flat 6 6 costar_flat_lt 9 6 costar_flat_l2 _6 costar flat_ 2 _6 costar flat 2 6 costar flat 2 _6_costar_flat_l2 6 costar _flat 12 6 costar flat 2 6 costar flat 2 A A C D1 E1 FA 1 A a T C D1 EEE aaa 6 gostar fatza 9 g a 00000000000 Q00000C Sov 0eg0e0ocnnn Soug000c g SrIg00eg0ecnndn Soug000 cc Sov 0e0ecnnnd o0g0go000ag0 O000G000000 PTeluleleletetereteters i Hi O O O O OOO D O oO 6 tar flat_ls 6 tar_flat_ 4 E oa aaa ae ac OOOO0900oggoG OODD CeCe er eres rene 2222000 il i F x Pick a comer Pick a direction ee cel gt Serpentine al 4 il b LF Sort Racks A Sorthy Grid Columns A Sorthy Grid Rows A Filter by Probe Head For Help press F1 610766 02 Page 63 of 225 Microlab STAR User Manual The sequence is re ordered across the racks as follows aj metl lay Hamilton Sequence Editor File Edit View Sequence Play Help ar AQ Re lIlnew JR REX O Taware _ _ costar flat _ _costar_flat_2 6 costar flat _6_costar_flat_l2 _6_costar_flat_lt 6 _6_costar_flat_l2 7 _6 costar_flat_lt 6 6 costar_flat_l2 9 6 costar_flat_lt _6_costar_flat_l2 di _6_costar_flat_
225. ware Editor from the Tool menu or 3 simply click on the Define Labware icon under on the tool bar If you want to add the new labware directly after definition choose the first procedure Whatever you choose the labware editor starts with the following main window N Untitled Hamilton Labware Editor File Edit View Help For Help press F1 17 2 Types of labware 17 2 1 Rectangular Racks and Plates Rectangular racks are specialized grids for holding either tips or containers in row and column order A microtiter plate is a rack in this sense and the wells represent the containers The rack is therefore a template describing a discrete number of positions for holding containers or tips Examples of racks include a tube rack a microtiter plate a microtiter strip a deep well plate and a tip rack The filename has the extension rck Page 184 of 225 610766 02 Microlab STAR User Manual NOTE JJ To change or define racks always use the Rectangular Rack and not the microplate definition within the labware editor 17 2 2 Containers Containers are vessels holding liquids e g the wells of a microtiter plate Containers are usually placed within racks Containers may be placed directly onto the carriers which is the case e g with reagent containers The filename has the extension ctr NOTE CO RE tips and needles are also defined as containers 17 2 3 Circular Racks C
226. wells i e aspirates liquids from wells on a plate and dispenses them to the corresponding wells on another plate A1 A1 H12 H12 The transferred volume will be 50 ul The method name is OnePlateToPlate The method uses new CO RE tips for every well We start with an empty target plate First an appropriate deck layout has to be created and saved as OnePlateToPlatePipette lay The deck layout for this method is shown in the picture on the next page 610766 02 Page 117 of 225 Microlab STAR User Manual G6 eon 24 30 35 40 45 Deck Layout for Method OnePlateToPlate To create this deck layout start the Deck Layout Editor by double clicking on the Microlab STAR Edit shortcut on the desktop Select New from the File menu Select ML_STAR as the current instrument if more than one instrument is installed Click OK Now you see the schematic view of the Microlab STAR deck Double click on one of the tracks A dialog box appears Select a standard carrier for microplates PLT CAR _L5MD from the ML_Star directory You may assign a name to the carrier Repeat this to place another plate carrier of the same type on to the deck Now add a tip carrier TIP_CAR_480 to the deck Double click on one of the sites of the plate carrier A dialog box appears Type in Source for the source plate and click Browse Select nun_96_fl_I rck from the Nunc directory Click OK Now
227. which may be achieved either by pressure or capacitive signal detection and transfer Loading unloading The process by which carriers are brought on to the work surface of the Microlab STAR and taken off again This can happen automatically by means of the Autoload Option or manually Method The method contains all instructions as to how the content of the source vessels is to be processed The assignment of the vessels happens virtually however they are positioned in the deck layout definition ML STAR User Software Software running Microlab STAR MTP Microtiter plate In general we assume a plate with 96 Wells 8 x 12 9 mm wide There are also plates with 384 Wells 16 x 24 4 5 mm or others with a different size Pause Interruption of processing The current processing steps are ended Page 212 of 225 610766 02 Microlab STAR User Manual Pipetting Transfer of liquids usually a defined number of aliquots from one container to another Pipetting arm Assembly consisting of at least 8 Pipetting channels as well as the common X driving and arm casing Pipetting channel Hardware part of the Pipetting arm which can be moved horizontally in the Y direction Pipetting module Firmware processor program which controls a pipetting channel in which category are included the Y and Z pipetting movement and the LLD Pooling Pipetting of different liquids in one well 1 2 3 ton and n to 1 2 3

- Frank`s Hospital Workshop

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