Home

USER MANUAL

1. Alternate DNS server Figure 23 Setting up your laptop to communicate with RoboSep 4 1 2 Set Up for Network Connection When we install your instrument RoboSep can be set up to automatically obtain an IP address from your network This will allow you to see shared folders on RoboSep from any other computer on the local network You may need to speak to your network administrator to see if this is permitted FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe i Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 33 0 4 76 18 99 63 www stemcell com e mail info stemcellfrance com Version 1 1 0 August 2005 Catalog 28940 4 1 3 Finding RoboSep on the Network Whether you are connected to a local network or to your laptop you will need to find RoboSep before you can access the shared folders If using Windows XP 1 Select Search from the Start Menu 2 Select Printers computers or people 3 Select Computer on network 4 Enter RoboSep name Robo XXXX where XXXX is the instrument serial number listed on the back of the instrument
2. Pipetting range 12 5 uL to 5 mL accuracy volume variations lt 10 for all the pipetting range Dimensions Height with lid 55 6 cm 21 inches Width 70 7 cm 27 inches Depth 39 2 cm 15 inches Weight 26 kg 57 Ib Power Requirements Electrical supply 100 120 220 240 V ac 50 60 Hz 260 W uses two 4A 250V 5 x 20 mm Type T fuses Conditions for Operation Temperature 10 C to 30 C Relative humidity 20 85 non condensing RoboSep is not specified to be used in a cold room 4 C 39 F For Indoor Use Certifications Pollution degree 2 Installation category Il Altitude 2000m CE and CSA certified RoboSep is intended for research use only 8 RoboSep Spare Parts and Accessories Product Name Catalog Additional One Year Warranty RoboSep Service Rack Hydraulic Fluid Bottle and Tubing RoboSep Buffer 250 mL EasySep RBC Lysis Buffer 10x Concentrate 100 mL RoboSep Tip Racks 1 box of 8 racks Ethernet Cross over Cable RoboSep Carousel The Big Easy EasySep Magnet FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version
3. 9 WV ANT AINE eer tea cages o a id A a 23 10 Contact Information a osasu cnau enwaa See eee a eee a ee ee 24 FOR RESEARCH USE ONLY ee E Gh ee OAT StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com ee al di en Verson 1 10 e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 e mail info stemcellgb com Catalog 28940 1 Introduction RoboSep is a laboratory instrument that fully automates EasySep immunomagnetic cell separation EasySep is a column free magnetic cell separation technology that enables both positive and negative selection of desired cells from virtually any species Using an integrated color touch screen RoboSep can be programmed to purify cells from up to 4 samples at once The user simply loads the correct consumables as indicated on the touch screen and presses Run RoboSep then automatically executes the EasySep immunomagnetic labeling and magnetic separation steps using disposable pipette tips RoboSep is intended for research applications only Please make sure that you have read the Safety Information Warnings section of this manual located on page ii before using the instrument 1 1 System description 1 1 1 EasySep Magnetic Cell Separation Technology EasySep is a powerful immunomagnetic cell selection procedure that combines the specificity of monoclonal
4. Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 www stemcell com Version 1 1 0 e mail info stemcellfrance com August 2005 Catalog 28940 Protocol Order The protocols are listed in order of last use with the most recently used protocols at the top of the list Figure 12 The protocols can also be sorted by name or type To change the basis of sorting touch the appropriate column heading For example to sort protocols in alphabetical order press the Protocol Name heading An arrow in the heading indicates if the list is sorted in ascending or descending order The user can move up and down the list by using the arrows at the top right corner of the protocols window 2 2 3 Loading the Carousel Once all the desired protocols have been selected you are ready to begin loading the carousel 1 Press the start button An image of the carousel appears Figure 14 The active quadrant number s is displayed at the bottom right of the window At the top center of the window the name of the protocol assigned to this quadrant is also displayed 2 Identify the necessary reagents and resources required for the separation On the left hand side of the window is a list of all the reagents and resources that need to be loaded for the current separation The selected separation protocol is listed just above the carousel and the quadrant number s for the protocol are listed along the bottom of the screen The user can move up and down
5. rica a Ree RS ee eo WE ee Pa we ee bk A A ee RD 6 1 3 Running RoboSep Quick Start Instructions ce rrenaren 4044 fees eee ebb eee eee eee oe eee ee ae 6 2 Running RoboSep Detailed Instructions 00 cee eee eee 6 2 1 INMtOGUCHON crac d eo ete and A eee Re Rates ARO LEER CREAR EEA ERO eR Mabe Paes BAG 6 2 2 Seting Up sa RUM sic do eb oa iho rd ode ein deis tddi ee et ae et v 2 21 Entermg a USEr ID arriba 7 2 22 Selecting PrOTOCO S ser s rada TA A A RA E A io 7 2 23 Loading the Carousel Vii A A A A AA A 8 2 2 4 Removing the Carousel viciosos A A A da 11 2 20 Propano CellS oe o E N wine ee RL A lee AA AAA AAA ie ee Say 11 2 2 6 Pause and Abort BUUONS etica rasos a a a E ee ie a Pa ees 11 2 3 Instrument Care and Shutdown 4 cierran sirt id da di de 11 2 3 1 Aseptic Operation of RoboSep y ici Didi a aa e iia pde 11 2 4 Repnin its si dd A A fled 12 3 RoboSep Protocol Editor o ooooccocnrcnnan a a a a a a 12 Sel Installation 2274 0254004405850 a A Ph Oe eed ee Oe ee a edd fo EA ETA 12 3 2 Stant UDi oe eaa ad iea edawae fate bee hoes kee by eee eae yak eta pede Wie bee Pa Ave bs bees 12 3 2 Editing an Existing PrOtOCO arcas hens Saeed kre Red ease PE Rn ede Bena dae eas 12 3 2 2 Waiting a New Protocol s ccace actebadetsoeGas utir beeen ee heed aaa aad eee dba gag Bathe 13 3 9 Protocol SUCIU subdesarrollado dd 14 3 3 1Protoco Specific FEAS seet itinere a Se ee e a dive Ed Wee Ea Sp ae de 14 3 3 2 Proto
6. the list by pressing the arrows located at the top and bottom of the list Lot numbers and sample IDs can be entered by first touching the item on the list and then the grey box under the Reagent Lot ID heading see below for further details 3 Load the reagents and resources The reagents and resources list starts with items near the center of the carousel and ends with items near the rim to facilitate aseptic loading Touch the first item on the list This highlights that item in green and indicates the location of the item on the carousel with a green circle Turn the carousel so that the indicated quadrant is in a convenient position and place the item in the indicated location Continue down the list loading each item in a similar manner 4 Before placing the tip rack in the carousel remove the plastic collar and then the top cap Figure 4 Ensure that the case does not open when removing the top cap Place the rack on the carousel and push down to fully seat the rack this ensures that the tips are in the correct location for pick up by the robotic arm 5 Loading Reagents The minimum reagent volumes required for the given cell separation procedure are displayed below the names of the required reagents Verify that your vial has that minimum required volume To assist in proper placement the reagent vial labels have symbols that correspond to symbols on the carousel and on the touch screen Please note that the EasySep magnetic particle vial s
7. where a quadrant defines the minimum functional area for processing a sample Each quadrant is designed to accommodate a 14 mL sample tube a magnet with a 14 mL separation tube up to 3 reagent vials a tip rack and 2 multifunctional 50 mL tubes Figure 5 The Robotic Arm is capable of vertical and rotational motion The stainless steel tip head at the end of the arm is used to pick up pipette tips Figure 6 The Carousel is divided into quadrants The quadrant shown here is fully loaded with the magnet tip rack reagent vials and tubes required to carry out a separation Figure 7 The tip head A has two conical surfaces that engage with the 1 mL B and 5 mL C sterile disposable pipette tips FOR RESEARCH USE ONLY By coordinating the rotational and vertical movement of the arm with the rotation of the carousel RoboSep picks up sterile disposable tips from the custom tip racks on the carousel and then uses them to transfer fluids from one container to another or to mix the fluids in a given tube The robotic arm can pick up both 1 mL and 5 mL sterile disposable pipette tios using a unique tip head that has two conical tip sealing surfaces Figure 7 This dual tip capability is required so that RoboSep can accurately dispense volumes from 12 5 uL up to 10 mL RoboSep uses 1 mL tips to aspirate and dispense the magnetic labeling reagents and 5 mL tips to transport the sample and buffers In
8. 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe StemCell Technologies Inc Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 www stemcell com Version 1 1 0 e mail info stemcellfrance com August 2005 Catalog 28940 Command Details rcubate Label incubate Cocktail and Sample Extension time s fz Incubation Time s fo Separate This command is functionally the same as the Incubate command and is used to set incubation times for magnetic separations The incubation time is the minimum amount of time before the next command in the sequence is executed Typical values for incubation time are between 600 and 900 seconds 10 and 15 min For extension time a minimum of 215 seconds is recommended Top Up Vial This command is used to top up a tube with EasySep Buffer before transferring the sample to the magnet for separation or after the supernatant has been removed following incubation in the magnet The main difference between a Top Up Vial command and a Transport command is that one uses the buffer tip whereas the other uses the cell tip Although the volume parameter appears to be available the Top Up Vial command uses the specified sample volume dependent working volumes for the protocol Command Details op Up Vial Label Top up Sample with EasySep Butter Extension time s fizo Source Butter Bottie
9. C Program Files STI RoboSep reports Subfolders are automatically created when necessary to organize the report files by year and then by month within the year s folders The files are in standard text format txt and are automatically named according to the following convention Report DDMMMYYYY txt To access these files you need to set up a connection between RoboSep and an external computer as explained in Chapter 4 Communications and File Transfer 3 RoboSep Protocol Editor The RoboSep Protocol Editor is a stand alone program that is installed on a personal computer The editor is used to modify existing protocols or to write new ones to then be imported into RoboSep M Note This program is not required for functionality of RoboSep 3 1 Installation 4 End of Run Report FEE HEHEHE Scheduled ETC Operator ID Actual Start Actual Finish Final Status Sample 1 Sample ID Sample Label Protocol ID Protocol Label Sample s Initial Quadrant Sample Volume Quadrant 1 Cocktail Particle Lysis Antibody Sample BCCM Waste Vial Separation Vial Sample Vial Tip Box Quadrant 2 Cocktail Particle Lysis Antibody Waste Vial Separation Vial Sample Vial Tip Box 200506137 13 29 29 53 13 Jun 2005 12 13 17 13 Jun 2005 12 26 08 COMPLETED 1 1220525925 Mouse T Cell Negative Selection 19751 4 5 9 ml 13569 0 69 ml 13256 0 69 ml Unl
10. Destination fot Sample Tube Volume l G Regie C rosore Proportion po Free Air Dispense 7 3 4 Saving a Protocol The Save option will not be available until the protocol is validated by the editor To do this click the Validate Protocol button located at the top right corner of the editor console If successful the symbol beside the Validate Protocol button will turn from a question mark to a check mark The editor will only ensure that all information that is critical for protocol execution is present validate rctoce D It does not ensure that your protocol is correctly programmed For example if a mix command is programmed for a reagent vial the 5 mL pipette tip will collide with the top of the reagent vial and will most likely cause an error during the run The validation process would not have warned the user of the event Validate Protocol FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom a e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com 3 5 Tran
11. EasySep nanoparticles without the additional magnetic field gradients provided by a column matrix Unlike larger microparticles used with other column free systems the EasySep nanoparticles do not interfere with subsequent FACS analysis and do not need to be removed In addition to the range of EasySep selection cocktails that are currently available to select specific cells other cocktails can be easily prepared using the flexible TAC technology Any mouse IgG anti cell antibody can be incorporated into TACs to select or deplete your cell of interest Prepare your own custom cocktail with Do It Yourself TACs or request a custom cocktail Plus virtually any cell type from any species can be selected using PE Biotin FITC or APC selection cocktails and your own appropriately labeled antibodies FOR RESEARCH USE ONLY Figure 1 Schematic drawing of EasySep magnetic labeling of human cells O Dil iai v gt b Anti Dextran j Cell e t 4 Tetrameric Antibody E a A Complex EasySep gt Magnetic s Nanoparticle TACs are comprised of a specific anti cell antibody which recognizes the target cell bound to an anti dextran antibody which recognizes the magnetic nanoparticle using a pair of rat anti mouse IgG linking antibodies Figure 2 EasySep procedure for positive selection Add EasySep selection cocktail Cell suspension Incubate a 15 minutes Add EasySep magne
12. North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom In Europe Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 Toll Free within e mail info stemcellfrance com United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com StemCell Technologies Inc www stemcell com Version 1 1 0 August 2005 Catalog 28940 1 1 4 The Tip Stripping Arm RoboSep replaces the disposable tips in the tip rack after each use This strategy eliminates the need for a waste tip area on the instrument and facilitates clean up after a run by simply disposing of the tip box appropriately Figure 8 shows how the pipette tips are removed the robotic arm positions the tips in the tip rack the tip stripping arm swings out to engage the tip head just above the pipette tip and then the robotic arm moves upward stripping the tip off the tip head The two different sized slots on the tip stripping arm engage the different sized pipette tips Figure 8 Disposable 5 mL tip being replaced in the tip rack The robotic arm places the tip partway into the rack A The tip stripping arm swings out to engage the tip head just above the tip B The robotic arm then moves up and the tip falls into the tip rack C 1 1 5 The Hydrauli
13. command execution can be changed A description of the commands is found in Section 3 3 Protocol Structure When satisfied with the edited protocol follow the procedure under Saving a Protocol to save your work Note In order not to over write the original protocol the Save As option must be selected instead of Save FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe i Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 33 0 4 76 18 99 63 www stemcell com e mail info stemcellfrance com Version 1 1 0 August 2005 Catalog 28940 3 2 2 Writing a New Protocol In the File menu click on New The boxes in the editor screen will become active and several fields will show flashing red exclamation points Figure 20 If you place the mouse over the flashing red points a context specific description will appear on the screen to assist with appropriate completion of the field Move Up amp Move Down Selected commands can be moved up or down to change the order in which they will be executed Protocol Type Select the type of separation fro
14. device such as a lamp Operating system boots up but RoboSep fails to fully initialize Power switch not left off long enough It takes up to 10 seconds for the electronics to fully reset after the power switch has been turned off After shutdown turn off power switch and count to 10 before turning the power back on RoboSep won t resume a paused run Lid up RoboSep will not run if the lid is in the raised position Lower lid and press resume on screen RoboSep won t shut down because you can t load the basic shutdown protocol Previous run not complete or needs to be unloaded Let run finish Press unload button on the Sample Processing screen Reagents left in tips RoboSep leaves residual volume in tips when pipetting small volumes This is normal operation and is required to meet the accuracy requirements for low volume reagent addition The residual volume is 20 uL or less Missed tip pick up Tip rack not placed properly on carousel Robotic arm or carousel was bumped during operation Make sure that tip rack is seated properly by pushing down firmly on the outside of the exposed portion Only operate with lid down Tips not well aligned with tubes Positional calibration is required Contact STI technical support Tip rack separates at snap joint when removing cap Excess force applied at this joint by mistake Ens
15. list and then enter the sample volume Repeat this process until all the desired protocols up to 4 are selected 3 Press Load to begin the loading sequence Lift the lid and load the indicated carousel quadrant s with the items listed on the left of the screen Press Loaded once each quadrant is ready When all quadrants are loaded close the lid 4 Press Run RoboSep will now complete the selected protocols unless it is paused or halted The time remaining until completion is displayed on the screen 5 Once the run is complete press Unload open the lid and collect the cells of interest For positive selections the cells of interest are in the magnet tube For negative selection the cells of interest are in the 14 mL sample tube of the second quadrant standard 2 quadrant negative selection or the 50 mL negative fraction tube of the first quadrant 1 quadrant negative selection 6 Unload the rest of the carousel and store the reagents appropriately Discard the tip boxes containing the used tips in the appropriate waste container 7 The instrument is now ready for another run If no more runs are planned for the day or if a spill occurred clean the instrument as detailed in Chapter 5 To conserve energy turn off the instrument at the end of the work day Note RoboSep protocols may differ from the standard manual separation protocols because they have been optimized for an automated system Unless otherwise n
16. on the touch screen may cause fluid to seep behind the external surface and into the electrical components Excess fluid should not pool at the margins of the touch screen Note RoboSep is resistant to spills but it is important to avoid spilling water inside the instrument through openings in the work surface such as around the carousel bearing column and the optical sensor opening If accidental spilling of biological material occurs in the carousel area of the instrument and a more thorough cleaning is needed Unload and remove the carousel Figure 18 Disinfect the carousel and magnets with dilute bleach solution and then rinse well with water Rinse with distilled water Disinfect the spill with dilute bleach solution Dispose of clean up material according to local regulations Rinse the work surface tub with distilled water Clean with 70 Ethanol 5 4 Replacing the Fuses The two fuses are located at the bottom right side of the instrument next to the power supply outlet and the power switch If RoboSep does not turn on when plugged in and the power switch is activated the fuses may need to be replaced To replace the fuses Disconnect the power cable from the power source Disconnect the power cable from the instrument Open the fuse box and examine the fuses Replace the faulty fuse s Always use Type T 5 x 20 mm fuses rated 4A 250V Reinsert the fuse box containing the new fuse s in its slo
17. or in the About tab of the user interface 5 Double click on the Robo XXXX icon You should now see the shared folders on RoboSep The shared folders are Protocols Reports and Logs 4 2 Adding Protocols RoboSep comes with pre installed protocols that have been optimized for cell separation from normal samples using supported EasySep products Protocols that you have created for other applications using the protocol editor see Chapter 3 must be copied into the Protocols folder on RoboSep Once you have found RoboSep on the network 1 Copy the RoboSep protocol s from your computer onto RoboSep by either copying and pasting or by simply dragging and dropping the protocol file into the Protocol Folder Note The file must have a unique name to avoid potentially overwriting existing protocols To secure the protocol against accidental modification change the file properties to read only 2 Once you have transferred protocols to RoboSep shut down the instrument completely and then restart This is necessary for RoboSep to register the new file in the protocol list 3 Verify that the protocol behaves as expected with at least a dry run Protocols that are no longer required can be removed from the protocol folder and stored in an archive sub folder on RoboSep or on your networked computer 4 3 Accessing Reports and Log Files RoboSep stores usage reports Section 2 4 as well as log files c
18. priming sequence at the beginning of every cell separation protocol to minimize the effect of air bubbles that naturally accumulate in the hydraulic system However when RoboSep is used for the first time or if the hydraulic fluid bottle has run dry it is necessary to run a separate priming protocol 1 Fill the hydraulic fluid bottle with 250 mL of deionized water and connect the bottle to the hydraulic system tubing in the compartment located at the right of the instrument Figure 10 2 If itis turned off switch the instrument on and wait until the end of the start up sequence 3 Press the Instrument Tasks tab select Basic Prime under the Maintenance tab and then press the green Load button 4 Place a 50 mL tube in the appropriate slot of the carousel as indicated on the screen 5 Press Run Repeat the priming sequence if no water has been expelled from the tip head into the waste tube by the end of the priming sequence Once priming is complete the waste tube can be emptied 1 3 Running RoboSep Quick Start Instructions Note Before using RoboSep always ensure that the hydraulic fluid bottle contains enough deionized water for the planned experiments 1 Power up RoboSep and wait for initialization to complete 2 Follow the instructions on the message bar at the top of the screen Press Select Protocol to bring up the list of available protocols Choose the desired protocol from the
19. separation using a second 5 mL tip keeps track of the appropriate incubation times A custom designed single use tip rack contains the pipette tips necessary to process a single sample Figure 4 The rack is sterilized by gamma irradiation Remove the cap prior to use Scheduling software sets the sequence in which RoboSep will execute all of the actions required to process up to 4 samples This scheduler gives RoboSep the flexibility to perform different types of separations at the same time Please refer to RoboSep Product information sheets PIS for sample volumes FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom ES e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com Figure 4 Tip rack with and without protective cap and lid 1 1 3 The Robotic Arm and Carousel The robotic arm Figure 5 rotates and moves up and down driven by 2 independent stepper motors The rotation of the carousel is driven by a single stepper motor The carousel is divided into 4 quadrants Figure 6
20. to 8 500 mL Human CD14 Positive Selecti Positive 3 31 2005 Review the information To remove the protocol press the X in the last column To change the protocol press the protocol name and repeat the protocol selection process above To change the sample volume while keeping the same protocol selected press the sample volume for this protocol and enter a new volume Repeat the protocol selection process until all the desired protocols are entered Up to 4 different protocols can be chosen When a negative selection protocol with 2 rounds of separation is selected the interface automatically assigns 2 consecutive quadrants for the protocol Once 3 quadrants are occupied only protocols using 1 quadrant will be displayed Human CD19 Positive Selecti Positive 3 31 2005 Human CD34 Positive Selectl Positive 3 2 2005 Human CD4 T Cell Negative Negative 6 8 2005 Human CD56 Positive Selectl i Positive 11 26 2004 Human CD8 Depletion Negative 3 31 2005 Figure 13 Entering sample volumes FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe StemCell Technologies Inc
21. which are executed in sequence 3 3 1 Protocol Specific Fields Protocol type Select Positive for positive selection protocols and Negative for negative selection protocols Maintenance and Shutdown protocols are for service and diagnostic routines and are not useful for cell separation Label The name you enter here is what will appear in the protocol list This name must be unique for it to be registered as a valid protocol Author Version Created and Modified These fields allow you to keep track of the history of a protocol that undergoes modification Sample Volume Mininum and Maximum These fields define the allowable range of sample volumes Minimum should be set such that all relative volume dispensing actions are of at least 12 5 uL For example if particles are added at 50 uL mL of cells then the minimum sample volume should be 250 uL 50 uL mL x 250 uL 1000 uL mL 12 5 yL Maximum should be set such that the total sample volume after all reagent additions is less than 10 mL For example if cocktail is added at 100 uL mL and particles at 50 uL mL of cells then the maximum sample volume should be set at 8 5 mL 8 5 mL 0 1 x 8 5 0 05 x 8 5 9 8 mL FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll
22. 0 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom a e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com 5 RoboSep Protocol Editor O x File Description Working Volume Threshold Enter the Validate Protocol D TM Protocol Type postive y sample threshold volume RoboSep ioe z will use the High volume for top up DES pt commands with any sample above or Author RoboSep User equal to the threshold volume The Low ee PA created Moite volume will be used for top up commands with any sample smaller than the threshold Sample Volume uL Minimum sn Maximum fi 0000 volume Working Volume Threshold uL ag lo E Command Sequence Working Volume Low and High Enter e Transport the lower volume and upper volume that ome RoboSep will use during top up commands Add Use the pull down menu to select the type of command to be added and click the add button Command Details Figure 21 The protocol editor screen working volume threshold fields 3 3 Protocol Structure A protocol is defined by protocol specific fields that define global parameters for the protocol and by a list commands
23. 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom gee e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com 9 Warranty Limited warranty to original purchaser RoboSep the Product as supplied and distributed by StemCell Technologies Inc STI and delivered new to the original purchaser is warranted by STI against manufacturing defects in materials and workmanship for the following limited warranty period one 1 year parts and labor This limited warranty begins on the date of installation and does not include transportation removal or reinstallation Warranty repairs must be performed by a STI authorized service center Transportation of the Product to and from the service center is the responsibility of the purchaser STI will repair or replace the Product at STI s option and at no charge with new or reconditioned parts if found to be defective during the limited warranty period specified above The Product must be returned during the warranty period with transportation charges prepaid to STI s designated service center Prior to returning any Product for warranty service the purchaser must contact STI for problem determination and service procedures STI does not warrant uninterrupted or error free operation of the product All replaced parts and Products become the property of STI and must be returned to STI Replac
24. 3 0 4 76 18 99 63 www stemcell com Version 1 1 0 e mail info stemcellfrance com August 2005 Catalog 28940 FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe i Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 33 0 4 76 18 99 63 www stemcell com e mail info stemcellfrance com Version 1 1 0 August 2005 Catalog 28940 StemCell Technologies Inc The Cell Experts www stemcell com In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 e mail info stemcellfrance com
25. Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom eae e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com Working Volume Threshold Low and High After the magnetic labeling steps RoboSep tops up the sample to a given working volume before transferring the sample to the magnet for separation The top up volume can be set such that sample volumes less than a given Working Volume Threshold are topped up to a Low working volume while sample volumes greater than or equal to the threshold are topped up to the High working volume This capability exists to improve separation performance for small volume samples Suggested settings for negative selection protocols are Working Volume Threshold 4000 uL Low 5000 uL High 10000 uL Suggested settings for positive selection protocols are Working Volume Threshold between 1500 and 2000 uL Low 2500 uL High 10000 yL 3 3 2 Protocol Command Specific Fields There are a limited number of commands used to automate cell separation processes on RoboSep These commands are Transport Mix Incubate Top up and Separate To add a command 1 Select a command from the pull down menu in the Command Sequence section and click Add 2 Enter the appropriate param
26. Laboratory Use Part 1 General Requirements UL Std No 61010 1 2nd Edition Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Part 1 General Requirements CONDITIONS OF ACCEPTABILITY The equipment is supplied with an approved power supply cord set or power supply cord with plug that is acceptable to the authorities in the country where the equipment is to be used FOR RESEARCH USE ONLY In North America Tel 604 877 0713 In the United Kingdom In Europe Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com www stemcell com Version 1 1 0 August 2005 Catalog 28940 Fax 33 0 4 76 18 99 63 e mail info stemcellfrance com FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe StemCell Technologies Inc Tel 33 0 4 76 04 75 30 Fax 3
27. The Fully Automated Cell Separator CATALOG 20000 USER MANUAL StemCell Technologies Inc ES The Cell Experts l www stemcell com FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe StemCell Technologies Inc Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 www stemcell com Version 1 1 0 e mail info stemcellfrance com August 2005 Catalog 28940 Safety Information Warnings This instrument is intended for research use only and must be operated by trained professionals To ensure adequate cooling of RoboSep place it on a clean flat surface and be sure that there is at least 10 cm 4 inches of clearance around each side of the instrument Make sure that the bench space is free of any foreign objects or debris that could block the fan vents Always connect the power supply to a 3 prong grounded AC outlet rated 4A 110 240 V using the AC power cord provided with RoboSep Do not use an adapter to a two terminal outlet Before plugging the instrument in be sure that the correct fuses are installed Use two 4A 250V 5 x 20 mm Type T fuses Electrical devices pose the r
28. abeled Not Used 13265 0 395 ml 326698 Unlabeled Needed Needed Needed Needed Not Used 10 0 ml Unlabeled Not Used Unlabeled Not Used Unlabeled Not Used Unlabeled Not Used Not Needed Needed Needed Not Needed Table 1 Example of report file To install the editor insert the CD and then double click on Setup For Release The installer will open and guide the user through the installation process 3 2 Start Up Launch the protocol editor from the shortcut on your desktop If this is not visible double click on ProtocolEditor exe in the default install location of the protocol editor of C Program Files STI RoboSep bin When the protocol editor console is initially opened all the fields are greyed out To begin you must select either New or Open from the File drop down menu Select New to begin creating a new protocol or Open to edit an existing protocol 3 2 1 Editing an Existing Protocol The quickest and most reliable way to create a protocol for a specific application is to base it on an existing protocol To edit an existing protocol with the protocol editor click the Open button in the File menu and select the desired protocol This will populate the editor screen with the commands and settings for that protocol Once the desired protocol is opened any of its settings can be changed commands can be added or deleted and the order of
29. ail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com 1 2 Instrument Set Up 1 2 1 Unpacking the Instrument A StemCell Technologies Inc authorized agent will be present to help you unpack and install the instrument 1 2 2 Instrument Placement To ensure adequate cooling of RoboSep place it on a clean flat surface and be sure that there is at least a 10 cm 4 inches clearance around each side of the instrument Make sure that the bench space is free of any foreign objects or debris that could block the fan vents RoboSep is designed to fit in most biocontainment hoods for sterile operation RoboSep is not specified for use in a cold room 4 C 39 F 1 2 3 Power and Data Connections Figure 10 Power and data connections hydraulic fluid bottle Connect the power cord provided with RoboSep to the power supply outlet located at the bottom right side of the instrument next to the fuse box and the power switch Figure 10 Connect the other extremity of the power cord to a 3 prong grounded AC outlet rated 4A 110 240 V Do not use an adapter to a two terminal outlet Data connections to RoboSep are through an Ethernet port and 2 USB ports Chapter 4 provides more detailed instructions on setting up RoboSep on a network or for connection to a laptop via cross over cable 1 2 4 Powering Up and Shutting Down Make sure the instrument is set up in an adequate location and is connecte
30. antibodies with the simplicity of a column free magnetic system EasySep can be used as either a positive or a negative selection approach to purify your cell of interest With EasySep positive selection highly purified cells are obtained by targeting the cells of interest with antibodies to a specific cell surface antigen These targeted cells are crosslinked to EasySep magnetic nanoparticles via StemCell s patented TAC technology Figure 1 in a standard FACS tube The tube is then placed directly in a specially designed EasySep magnet This handheld magnet is gently inverted to remove the cells that are not bound to the magnetic nanoparticles To obtain a highly purified cell population the cells remaining in the tube are resuspended and separated twice or more then easily collected by removing the tube from the magnet Figure 2 For negative selection applications unwanted cells are targeted for depletion using a cocktail of monoclonal antibodies directed against specific cell surface antigens The labeled unwanted cells are then crosslinked to EasySep magnetic nanoparticles in a standard FACS tube The tube is placed directly in the specially designed EasySep Magnet This handheld magnet is gently inverted to remove the desired cells which are not bound to the magnetic nanoparticle complexes The unique EasySep magnet generates a high gradient magnetic field in the interior cavity that is strong enough to separate cells labeled with
31. ation please contact USA and Canada 1 800 667 0322 email info stemcell com Other regions 1 604 877 0713 email info stemcell com Head office StemCell Technologies Inc 570 West Seventh Avenue Suite 400 Vancouver BC V5Z 1B3 Canada FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom gee e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com e o CSA INTERNATIONAL Certificate of Compliance Certificate 1660865 Master Contract 227130 Project 1660865 Date Issued 2005 04 04 Issued to StarFish Medical 555 Ardersier Rd Unit 5 Victoria BC V8Z 1C8 Canada Attention Mr Fabio De Pasquale The products listed below are eligible to bear the CSA Mark shown with adjacent indicators C and US Issued by Stephane Poutissou P Eng Authorized by Ken Rutledge Product Group Manager MJ PRODUCT CLASS 872104 LABORATORY EQUIPMENT Electrical CLASS 872184 ELECTRICAL EQUIPMENT FOR LABORATORY USE Certified to US Standards Blood Separator Model RoboSep rate
32. c System The hydraulic system is comprised simply of a positive displacement piston pump connected to a deionized water bottle on one side and to the pipette handling tip head on the other The only visible parts of this system are the water bottle and the tubing running from the console to the robotic arm The positive displacement piston pump does not require any user maintenance because the piston seal has a substantially longer operating life than the seals on syringe pumps used in other instruments In addition because the hydraulic fluid does not contact the samples or reagents RoboSep uses deionized water which is less damaging to the piston seal The use of water instead of air as a hydraulic fluid allows a high degree of accuracy and high speed dispensing The water effectively acts as an incompressible extension of the pump piston and therefore the hydraulic system operates with a dead air volume similar to that in a manual pipettor 1 1 6 Homing the Carousel and Robotic Arm RoboSep tracks the position of the robotic arm and carousel relative to physical markers called homing flags A home position is registered by the computer when a homing flag trips an optical sensor The carousel homing flag is readily visible as a projection on the bottom plate This projection trips an optical sensor which can be seen on the front inside wall of the bowl when the carousel is removed The robotic arm homing flags and sensors are not visible Homing o
33. col Command Specihe FIGS vecina da dt Paid a taa ic gz 15 BA Saving a Protocol ivi A A A A A A A A oe i 17 FOR RESEARCH USE ONLY rena Ta Aaa Tees Bi azena zean E T cs e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com 4 Communications and File Transfer 00 cece 18 4 41 Connecting to RObGSe OM i is edie a a A a A ies Bea 18 4 1 1 Set Up for Cross Over Cable Connection 0 0 0 0 0 cette 18 4 12 Set Up Tor Network CONNECTION eres wide paa bbe ee e e be a ebb he ed a ee antes 18 4 13 Finding Robosep on the Network ssa ie teehee a eed eed Led pie pe ee eee 19 4 2 AQUINO Protocols iG eas ots Baca Ra ee aes Ath ae A ates ese ad ee Ra eel dade pels wee eee 19 4 3 Accessing Repons and Log FES s serorei piine aean aoa vee ee Ee aed oe ee gw ane a EA 19 5 Care and Maintenance 000 cee es 20 Bal Generali CONSIGSPALONS viril a Gath ate RA eno emis eee acne idan ooh adel oa eee eae Lee 20 9 2 Daily Maintenance rca 0c ene dean a ee Da Nees Dt BR eRe ta eee eed 20 5 3 Cleaning RObOSe6p seri kecin een rior edo hdl dds ds gtd Abia we nb bated ede eee de ee 20 5 4 Replacing the FUSES iii a eee ae heen GA ened ee each Desh oe eh ee end ce atta eit made eee 20 6 Troubleshooting css cresa nnsa a ee ee eee ee ee ee ae 21 7T SDCCWICANGNS essers tease de teen AU A A eh ea eben de a E 22 8 RoboSep Spare Parts and Accessories 0000 cee eee 22
34. cubate This parameter specifies the length of time in seconds for reagent and magnetic separation incubations FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom a e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com Description Validate Protocol gt Protocol Type Positive Label lumen 03 pe Mahor fRoboSepuser SSS Version Created Sample Volume uL Minimum 250 Maximum 10000 Working Volume Threshold uL tooo Low 5000 High roooo Add Transport v Delete Transport Human CD34 Cocktail Command Details Transport Human CD34 Cocktail Extension time s 420 Source fat Antibody Cocktail Vial Destination fot Sample Tube Volume Mo Relative C Absolute Proportion 0 10 Transport Label Free Air Dispense rl Figure 22 Protocol Editor Screen with a new protocol open The first transport command has been entered Transport This command is used for transporting fluid from one location on the carousel to another and is used in most protocols to
35. d 100 120 220 240 V ac 50 60 Hz 260 W Note The above model is Equipment Class I Pollution Degree 2 Installation Category II The C and US indicators adjacent to the CSA Mark signify that the product has been evaluated to the applicable CSA and ANSI UL Standards for use in Canada and the U S respectively This US indicator includes products eligible to bear the NRTL indicator NRTL i c National Recognized Testing Laboratory is a designation granted by the U S Occupational Safety and Health Administration OSHA to laboratories which have been recognized to perform certification to U S Standards DQD 507 Rev 2004 06 30 FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom os e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com ice e CSA INTERNATIONAL A A A Certificate 1660865 Master Contract 227130 Project 1660865 Date Issued 2005 04 04 APPLICABLE REQUIREMENTS CAN CSA C22 2 No 61010 1 04 Safety Requirements for Electrical Equipment for Measurement Control and
36. d to a power source as described in sections 1 2 2 Instrument Placement and 1 2 3 Power and Data Connections Power up RoboSep by activating the switch located at the bottom right side of the instrument Figure 10 RoboSep first loads the computer operating system and then the instrument control software and user interface The touch screen provides visual confirmation that these processes are occurring After a short period of time the robotic arm and carousel will move The start up sequence is complete once the start screen is visible see Figure 11 To shut down RoboSep M go to the Instrument Tasks section of the graphical user interface by pressing the appropriate tab at the top of the touch screen Press Basic Shutdown Protocol Wait until the screen becomes black and then turn off the power switch FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 e mail info stemcellfrance com StemCell Technologies Inc www stemcell com Version 1 1 0 August 2005 Catalog 28940 1 2 5 Priming the System RoboSep performs a
37. dragging and dropping Close all windows Double click on the Start RoboSep icon on the desktop to re open the user interface Note It is possible to import protocols without exiting the user interface However the imported protocol will not appear in the Select protoco P list until the user interface is re started 4 Communications and File Transfer RoboSep runs on Window XP and as such has many possibilities for communication with other computers RoboSep comes configured for communication with your laptop via a cross over cable supplied but can be configured to be connected to your network 4 1 Connecting to RoboSep For network access to shared folders login as guest on the RoboSep machine FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom ae e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com 4 1 1 Set Up for Cross Over Cable Connection To configure your laptop go to the Change the Local Area Connection Properties Table by selecting Network Co
38. e e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com Minimum reagent volume required Scroll button for reagent list Figure 14 Carousel layout The locations of the items listed along the left hand side of the screen are indicated with a green circle when the listed item is selected In this example the waste tube is highlighted on the left and indicated with a green circle in quadrant 2 Sample Processing Reagents and Resources The sample tube and separation tube are 14 mL round bottom tubes e g Falcon 2057 The waste tube and negative fraction tube lysis buffer tube are 50 mL conical bottom tubes e g Falcon 2070 The Service Rack is provided to hold the reagents and sample prior to loading them on the carousel and to keep track of the caps during a run Figure 16 The rack is laid out in quadrants like the carousel with symbols on the reagent storage locations to assist with keeping track of the correct caps Human CD4 T Cell Negative Selection 19052 Quadrant s 3 Run Progress Time remaining 00 50 Figure 15 Run Progress Once a run has begun progress is shown at the bottom left with 1 an indication of how much time remains FOR RESEARCH USE ONLY Waste Tube Separation Tube Negative Fraction Lysis Buffer Tube Sample Tube Tip Rack EasySep Reagent Vials I
39. e mail info stemcell com Tel 0800 731 27 14 e mail orders stemcell com Fax 0800 731 27 13 e mail info stemcellgb com StemCell Technologies Inc www stemcell com Version 1 1 0 August 2005 Catalog 28940 2 2 4 Removing the Carousel The carousel can be easily removed from the instrument for cleaning or to facilitate loading and unloading of the samples and reagents Some users may prefer to remove the magnets from the carousel prior to removing the carousel as the magnets are relatively heavy To remove the carousel 1 Rotate the black central knob Figure 18 counterclockwise until the threads have disengaged To replace the carousel within the instrument 1 Place the carousel over the bearing hub 2 Align the holes around the knob with the pins on the bearing hub by slowly rotating the carousel 3 Screw the central handle clockwise until locked It is important to tighten the knob fully so that the carousel Figure 18 Close up of knob that fastens the carousel to the bearing hub stays level when the robotic arm pushes down on it to pick up pipette tips 2 2 5 Preparing Cells Refer to the product information sheet that comes with your EasySep reagents for instructions on how to prepare cells for separation using RoboSep It is important to use the recommended cell concentration for best results Initial cell suspension volume should be between 250 uL and 8 5 mL Ideally the sample should be a single cell suspensio
40. e the 14 mL tube containing the isolated cells from the magnet Figure 17 A o If desired collect the 50 mL tube containing the depleted fraction located at the left of the tip box Negative selection o If two rounds of separation were performed collect the enriched cells in the 14 mL tube located at the left of the magnet on the second quadrant used for the negative selection procedure Figure 17 B o If one single round of separation was performed collect the 50 mL tube containing the enriched cells located at the left of the tip box Figure 17 C o If desired collect the 14 mL tube s containing the labeled cells located in the magnet s Unload the carousel and store the reagents appropriately Discard the tip boxes containing the used tips in the appropriate waste container Check that the hydraulic fluid bottle contains enough deionized water for further experiments The instrument is now ready for another experiment FOR RESEARCH USE ONLY Selected cells Figure 17 Location of desired cells after separation A Positive selection B Negative selection 2 rounds C Negative selection 1 round In North America In the United Kingdom In Europe Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Toll Free Fax 1 800 567 2899 United Kingdom
41. ement parts and Products assume the remaining original warranty or ninety 90 days whichever is longer This limited warranty covers manufacturing defects in material and workmanship encountered in normal use of the Product and shall not apply to the following including but not limited to damage which occurs in shipment application and uses for which the Product was not intended failures or problems which are caused by Products or equipment not supplied by STI accidents misuse abuse neglect misapplication fire water lightning or other acts of nature incorrect electrical voltage fluctuation or surges damage caused by improper or faulty installation improper connection with any peripheral Product alteration or modification improper or unauthorized repair cosmetic damage or exterior finish failure to follow operating instructions customer adjustments maintenance and environmental instructions that are covered and described in the user manual use of non STI or unauthorized parts supplies accessories or equipment which damage this Product or result in service problems failure or problems due to incompatibility with other equipment STI is not liable for any damage to or loss of any data or other information stored on any media contained within the Product or any non STI product or part not covered by this warranty Recovery or reinstallation of data or other information is not covered under this limited warranty In no e
42. eters see below 3 Click on the white area in the Command Sequence section to confirm the settings Each command has specific parameters that influence how it is executed These will appear in the Command Details section of the editor console All commands have a Label parameter and an Extension Time parameter The Transport Mix and Top up commands have Source and Destination parameters The Transport and Top up commands have the Free air Dispense and Volume parameters The Incubate and Separate commands are identical and have the Incubation Time parameter Label The description in this box will appear in the Command Sequence list once all the details of the command have been entered Extension time Each command takes a set amount of time to execute The extension time specifies what additional time is available to RoboSep to facilitate the scheduling of all the required actions Source This is the container tube or reagent vial from which the pipette will aspirate If a reagent vial is specified as the source a 1 mL pipette tip is used automatically If a sample tube or magnet tube is the source then the 5 mL tip reserved for cell handling is used If the source is the buffer bottle or lysis buffer tube negative fraction tube then the 5 mL tip reserved for buffer handling is used The source is chosen from a pull down list which is sorted by relative quadrant nu
43. f the carousel and robotic arm is performed automatically at start up at the beginning of each cell separation experiment and after the instrument has been paused during a run The robotic arm home positions are detected again periodically throughout a run To ensure correct operation of the instrument it is essential that no mobile part be moved or touched during a run Homing is the only mechanism by which RoboSep determines where all the items on the carousel are and so any disruption of the carousel or the arm while they are in operation will likely cause pipetting actions to fail This is why RoboSep operates with a transparent lid over the arm and carousel If the lid is opened during an experiment the instrument will finish the current action and then pause RoboSep will automatically execute a homing sequence once the lid is closed and the run is resumed Figure 9 The carousel home flag is visible as a projection from the bottom plate in this schematic diagram FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom eae e mail info stemcell com Tel 0800 731 27 14 August 2005 e m
44. is has been verified by particle count as per ISO standard 14644 1 Although the hydraulic fluid is internal to the instrument and does not come into contact with the sample reagents or tips during normal operation we recommend using sterilized water for aseptic operation FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom a e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com 2 4 Reporting Usage data is automatically saved during RoboSep operation RoboSep has the ability to keep track of the experiments that are performed using the instrument To benefit fully from this function it is recommended that the following information be provided at the time of the experiment User ID in the case of multiple users Lot number for each of the reagents used with the instrument Instructions on how to input this data are detailed in Chapter 2 Running an Experiment Detailed Instructions One report file is generated for each day when RoboSep is active The files can be found in the folder
45. isk of electric shock To reduce the risk of shock do not open any covers that are fastened with screws While RoboSep is designed to withstand spills on its exterior surface do not allow fluids to enter the interior of the instrument In the event of such a spill disconnect the power cable before cleaning up Do not operate RoboSep in extreme humidity or in conditions that can create condensation Protect the instrument against dust and moisture and avoid physical shock and strong forces Do not exert excess pressure on the screen The EasySep magnets on the RoboSep carousel generate strong magnetic fields Keep away from computer discs pacemakers watches and other objects that respond to magnetic fields Always operate RoboSep with the lid down In case of malfunction call StemCell Technologies Inc for service Contact information is located at the back of this manual Do not attempt to fix the instrument yourself Never remove the outer casing of RoboSep There are no user serviceable parts inside the instrument Notice RoboSep is designed and certified to meet UL CAN CSA CENELEC EN 61010 1 safety requirements Safety certified products are safe to use when operated in accordance with the user manual This instrument should not be modified in any way Alteration of this instrument will Void the manufacturer s warranty Void the 61010 1 safety certification Create a potential safety hazard This product conf
46. ize for RoboSep has increased to 1 5 mL Use of the 0 5 mL EasySep vial may cause the RoboSep pipette tip to jam 6 Press the green Loaded button when all the reagents and resources are loaded on the active quadrant s The reagents and resources required for the next selected protocol will appear 7 Repeat steps 2 6 until all the required quadrants are loaded Close the instrument lid 8 Press the green Run Figure 14 button to start the cell separation experiment A warning message appears asking the user to verify that all the reagents and resources are correctly loaded and that nothing can interfere with the movements of the instrument Please ensure that the lids have been removed from all vials and resources especially the buffer bottle If everything is clear press Confirm 9 Observe that the initial prime step ejects water from the tip head into the waste tube in carousel quadrant 1 This ensures that the hydraulic fluid system is properly connected The experiment is now in progress The time remaining until completion is displayed at the bottom of the screen Figure 15 FOR RESEARCH USE ONLY Tel 604 877 0713 Tel 144 0 20 7837 7565 Te 93 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom ea
47. m the pull down menu Label Enter the name of the protocol that will appear in the Protocol Selection screen on RoboSep M Author Enter the author of the protocol Sample Volume Enter the minimum and maximum sample volumes Command Sequence The list of programmed commands will appear here in the order in which they will be executed Command Details Parameters that are specific to different commands can be changed here Move Up amp Move Down Selected commands can be moved up or down to change the order in which they will be executed RoboSep Protocol Editor Protocol Type Select the type of separation from the pull down menu Label Enter the name of the protocol that will appear in the Protocol Selection screen on RoboSep Modified Maximum fo High Author Enter the author of the protocol Sample Volume Enter the minimum and maximum sample volumes Command Sequence The list of programmed commands will appear here in Command Details the order which they will be executed Command Details Parameters that are specific to different commands can be changed here Figure 20 The protocol editor screen fields Required information is highlighted with an exclamation mark FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 3
48. mber The source tube should be in Q1 in most cases since Q1 is the start quadrant for all protocols This means that a protocol loaded in quadrant 2 will start in quadrant 2 Destination This is the container into which the pipette will dispense The destination container is also selected via a pull down menu The list is sorted by relative quadrant number A protocol with a source in Q1 and a destination in Q2 will occupy 2 quadrants on the carousel If the protocol is loaded in quadrant 3 the destination will be in quadrant 4 Free air Dispense When this check box is enabled RoboSep dispenses the fluid that it is transporting just below the upper lip of the destination container and above the liquid in the container This option is useful where contact with the contents of the destination tube is undesirable e g preventing contamination of the buffer tip when adding buffer to the sample tube Note This option is not recommended for transfer of small volumes e g addition of reagents Volume For Transport commands the user can specify whether RoboSep is to transport an absolute volume in uL or a volume relative to the initial sample volume e g To have RoboSep add 100 uL of reagent per mL of sample volume program the addition of a relative volume with a proportion of 0 10 This parameter is not enabled for Top up commands since the top up volume is dictated by the sample working volume In
49. n However RoboSep tips will not clog if small aggregates are present 2 2 6 Pause and Abort Buttons Hitting the Pause button will cause RoboSep to interrupt the run as soon as it completes the current action Pressing the Resume button will restart the run at the next scheduled action after RoboSep orients itself by performing homing actions The Abort button also stops execution of the run as soon as the current action is complete but with the option to exit from the protocol completely or to resume execution Instrument Tasks 2 3 Instrument Care and Shutdown If no more experiments are to be performed on that day or if there was a spill on the instrument clean the instrument as detailed in the Care and Maintenance section Chapter 5 To conserve energy turn off the instrument at the end of the work day Navigate to the Instrument Tasks section of the graphical user interface Figure 19 Select Basic Shutdown Protocol Wait until the screen becomes black and then turn the Figure 19 The shutdown protocol is located under Maintenance on the Instrument Tasks tab switch on the right side of the instrument to the off position 2 3 1 Aseptic Operation of RoboSep To ensure aseptic operation RoboSep must be used in a biocontainment hood RoboSep is designed to fit easily in most commercially available hoods In addition RoboSep does not affect sterility within the biocontainment hood Th
50. n North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 e mail info stemcellfrance com StemCell Technologies Inc www stemcell com Version 1 1 0 August 2005 Catalog 28940 Sample ID and Lot numbers Sample ID and reagent lot numbers can be entered during the loading sequence for each protocol 1 Select the sample or reagent by touching the button in the list on the left of the screen 2 Touch the grey box under the Reagent Lot ID heading in the upper right corner of the screen This brings up a numeric key pad 3 For reagents enter the last 5 digits of the reagent lot number for sample ID up to 48 digits can be entered Note Although EasySep reagent lot numbers are formatted 88A88888 only the last 5 digits are required to uniquely identify the lot Figure 16 The Service Rack is divided into quadrants like the carousel Collect the cells at the end of the experiment Press Unload and allow 5 seconds for the robot and carousel to move to a safe position Open the instrument lid Collect the cells of interest Positive selection o Remov
51. nnections in the Control Panel Select Internet Protocol TCP IP and hit the Properties button to bring up the properties screen Figure 23 1 Select Use the following IP address 2 Set IP address to 192 168 1 201 3 Set Subnet Mask to 255 255 255 0 Search 1 Folders E 4 Local Area Connection Properties le E Go Device Name Phone or Host Address A General Authentication Advanced Connect using Internet Protocol TCP IP Properties B Intel R PRO 100 VE Network Conne i General You can get IP settings assigned automatically if your network supports this capability Otherwise you need to ask your network administrator for the appropriate IP settings This connection uses the following items B QoS Packet Scheduler 36 AEGIS Protocol IEEE 802 1 v2 3 1 7 2 Internet Protocol TCP IP Obtain an IP address automatically Use the following IP address IP address 10 0 0 100 Subnet mask 255 255 255 0 Q a Install Uninstal Description Transmission Control Protocol Internet Protocol The default wide area network protocol that provides communication across diverse interconnected networks Default gateway Use the following DNS server addresses C Show icon in notification area when connected Notify me when this connection has limited or no connectivity Preferred DNS server
52. ontaining more detailed diagnostic information One report file is generated for each day that RoboSep is run Log files may be useful to an STI designated service technician to assist in troubleshooting One log file is started every time RoboSep is started To access report files Go to the reports folder and then go to the appropriate year and month to view the reports listed by date The files have the format Report_ DDMMMYYYY txt To access log files Go to logs folder The files have the format robosep log XXX where XXX txt for the most recent log file 1 for the next most recent and so on To identify the log file corresponding to a given run it is necessary to know the time and date of that run The precise information is available in the corresponding report file FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe StemCell Technologies Inc Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 www stemcell com Version 1 1 0 e mail info stemcellfrance com August 2005 Catalog 28940 5 Care and Maintenance 5 1 General Considerations RoboSep is designed to avoid any c
53. orms to the Class A standards for electromagnetic emissions intended for laboratory equipment applications according to FCC Part 15 and CENELEC EN 61326 It is possible that emission from this product may interfere with some sensitive appliances when placed nearby or in the same circuit as these appliances The user should be aware of this potential and take appropriate measure to avoid interference StemCell Technologies Inc is not responsible for any injury or damage caused by the use of this instrument for purposes other than those for which it is intended or by modifications of the instrument or software not performed by StemCell Technologies Inc or an authorized agent FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom a e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In
54. oted all RoboSep procedures have been optimized for use at room temperature regardless of the temperature recommended for the manual procedure Sample Processing A Messages Press Select Protocol to choose a separation protocol 2 Running RoboSep Detailed Instructions Samples 2 1 Introduction Power up the instrument as indicated in Chapter 1 2 4 Powering Up and Shutting Down The Robosep graphical user interface will load automatically Wait for the end of the start up sequence before touching the touch screen the lid or other moving parts of the instrument Start up is complete when the Sample Processing screen appears on the user interface screen as shown in Figure 11 Start screen Run Progress Time remaining User ID Figure 11 Start screen FOR RESEARCH USE ONLY In North America In the United Kingdom In Europe i Tel 604 877 0713 Tel 44 0 20 7537 7565 Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 604 877 0704 Fax 44 0 20 7515 5408 Fax 33 0 4 76 18 99 63 www stemcell com Toll Free Tel 1 800 667 0322 Toll Free within e mail info stemcellfrance com Version 1 1 0 Toll Free Fax 1 800 567 2899 United Kingdom ae e mail info stemcell com Tel 0800 731 27 14 August 2005 e mail orders stemcell com Fax 0800 731 27 13 Catalog 28940 e mail info stemcellgb com The user interface consists of 2 main sections defined by tabs at the top of the screen the Sam
55. ple Processing section that is visible at start up and the Instrument Tasks section Figure 11 The light grey tab indicates which section is currently active You can switch from one section to the other at any time by touching the appropriate tab The various functions accessible in these 2 sections are detailed in the following chapter 2 2 Setting Up a Run Go to the Sample Processing section of the user interface The sample processing screen is initially divided into 5 areas Messages Samples Run Progress User ID and a row of action buttons along the bottom The Messages area indicates what step is next in the set up process to start running RoboSep The Samples area is divided into 4 rows corresponding to the 4 quadrants on the carousel Each row has columns to display the protocol name the sample volume mL and the type of cell selection positive or negative Once protocols have been selected this area indicates on which quadrant of the carousel each protocol will be run Figure 12 The Run Progress area has a progress bar and count down clock that show the time to completion of the run Figure 15 The USER ID area allows the user to enter their unique code for reporting purposes see Table 1 Section 2 4 The action buttons are Abort Pause and Load 2 2 1 Entering a User ID To enter a user ID 1 Press the grey box under the User ID heading A numeric pad will pop up 2 Key in the user ID and p
56. ress the Enter key A user ID can be input at any point before a run begins The User ID information is stored internally and is included in the report file that is generated upon completion of each run see Table 1 Section 2 4 User ID helps to keep track of usage in a multi user environment Sample Processing Messages Choose a protocol There are 4 available quadrants Protocols wv Human CD14 Positive Selection 18058 1 Positive 3 31 2005 Human CD19 Positive Selection 18054 Positive 3 31 2005 Human CD34 Positive Selection 18056 Positive 3 2 2005 Human CD4 Positive Selection 18052 Positive 11 26 2004 Human CD4 T Cell Negative Selection 19052 Negative 6 8 2005 Human CD56 Positive Selection 18055 Positive 11 26 2004 Human CD8 Depletion Negative 3 31 2005 2 2 2 Selecting Protocols To select a protocol 1 Touch Select Protocol in any row of the Samples area A list of the available protocols appears on the screen Figure 12 2 Select a protocol by touching the protocol name A numeric pad pops up Figure 13 8 Key in the sample volume in mL and press the Enter key The original Sample Processing screen will re appear with the selected protocol in the appropriate quadrant row Figure 12 Select a protocol from list Sample Processing Messages Choose a protocol There are 4 available quadrants Protocols Sample Volume E Enter a volume in the range 0 250 mL
57. ross contamination of the samples or reagents manipulated during the experiments In addition RoboSep can be operated if desired within a biocontainment hood to maintain sample and reagent sterility To ensure that RoboSep performs as expected regarding these aspects it is the responsibility of the operator to maintain a clean and uncluttered environment When using RoboSep as during any research experiment it is essential to observe good laboratory practices as described by your institution or company 5 2 Daily Maintenance 1 Wipe down all exposed surfaces with an alcohol swab 2 Verify that the hydraulic fluid bottle is full 3 Wipe down the surfaces of the stainless steel tip head that hold the pipette tips Figure 7 This tip head can accumulate a thin film of plastic from repeated loading and unloading of pipette tips that will degrade the ability of the tip head to seal adequately with the pipette tips 5 3 Cleaning RoboSep The preferred method to clean and disinfect RoboSep is to lightly spray 70 Ethanol or 70 Isopropanol on the instrument s external surfaces and wipe it off with a soft and clean paper towel RoboSep can also be wiped down with a dilute bleach solution e g 1 in 10 dilution of 5 sodium hypochlorite solution but should then be wiped clean with water For the touch screen it is recommended that the 70 isopropanol be sprayed on a soft and clean paper towel prior to wiping down Excessive direct spray
58. sferring protocols onto a USB key 1 2 3 Insert your USB key into the USB port on your PC Your PC will detect the USB key as a drive storage device Copy the RoboSep protocol s onto the key by either copying and pasting or by simply dragging and dropping the protocol Remove your USB key from the USB port 3 6 Importing protocols from a USB key onto RoboSep 1 To get to the desktop from within the user interface connect a mouse and keyboard to the USB ports on the right side of RoboSep and press the Windows button located between Ctrl and Alt This should give you access to the Windows taskbar where you can exit of the user interface by closing the DOS window Open the RoboSep folder by double clicking the Shortcut to RoboSep icon on the desktop If the Shortcut to RoboSep icon is not present on the desktop use the path C program files ST I RoboSep after double clicking My Computer Next double click on the protocols folder Insert your USB memory key into either one of the USB ports on RoboSep If you have both a mouse and keyboard plugged into RoboSep at this time it is recommended that the USB memory key be inserted in place of the keyboard the mouse will be needed to drag and drop the protocols Copy the protocol from your USB key into the protocols folder by either copying and pasting or by simply
59. specify the amount of reagent to add to the sample and to specify the volume of sample to move from one tube to another Typically reagent volume addition is relative to the sample volume For transporting the sample from the sample tube to the separation tube specify the maximum volume and RoboSep automatically adjusts to transfer the whole amount Mix This command is used to mix the contents of a tube This command uses 5 mL pipette tips and therefore should only be executed where the contents to be mixed are in a 14 mL or 50 mL tube The source and destination tubes should be the same There are no other parameters to specify since the mixing regime is preset as a function of the volume present in the tube Command Details Mx Label Mix Cocktail and Sample Extension time s 120 Source 21 Sample Tube Destination E Sample Tube y Incubate This command is used to set incubation times for reagents The incubation time is the minimum amount of time before the next command in the sequence is executed Typical values for incubation time are between 600 and 900 seconds 10 and 15 min For extension time a minimum of 215 seconds is recommended FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20
60. t Press gently until it snaps into place Reconnect the power cable to the instrument then to a grounded power source FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe i Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 33 0 4 76 18 99 63 www stemcell com e mail info stemcellfrance com Version 1 1 0 August 2005 Catalog 28940 6 Troubleshooting RoboSep will present error messages to help diagnose some types of problems that may occur during use Please record the message before calling technical support Section 10 Contact Information The following tables address other potential issues that are not flagged as errors Problem RoboSep screen does not turn on No sound movement or sign of activity has been noticed 30 seconds after powering up the instrument fails to Potential Cause Faulty fuse Solution Check that the fuses are correctly installed as explained in the Care and Maintenance section of this manual Replace fuses start up No power to outlet Check that outlet is active by connecting a known working
61. the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe StemCell Technologies Inc Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 www stemcell com Version 1 1 0 e mail info stemcellfrance com August 2005 Catalog 28940 IV Table of Contents 1 Introduction is ceed eee eee oneness A wees Ree wee ete chon es 1 11 SYSTEM IDESCHOUGN oe 2a ux ace tok Pm Uo ele ue Coton eh aes BARR ane ER Ge hh Bae dh ONE eee AE Aa Sees Rhee 1 1 1 1 EasySep Magnetic Cell Separation Technology 6 ee cee nee e enn e eens 1 1 1 2 Robotic SYSTEM Overniew cia fie ha e ee ae Race bec Huei A Pee We ee are eed 2 121 3 The Robotic Anm andi Carousel i a ga 4 ge ai a a Rake da oe 3 11 4 The TESTERNA sss stare A A Barz ees E A ARA iS Boa tian WE RS 4 1 15 The Mydrauli Systemi viera eee Ye ged hos AA bee wa naw a dae bebe dd oad 4 1 1 6 Homing the Carousel and ROBOG AMT reccsent oare ee ioe ee ee ee be eae ee a a 4 12 Instument oct Up nit a A A E a a Data 5 1 21 Unpacking the INStUMEE 200 tee A E eee RAE eRe ee ee 5 1 22 Instrument Place MENE iia php Peewee bee eed dd goatee ddd a pe td dea ee be eed 5 1 2 3 Power and Data CONMECUIONS ipods deen Dir vie GA ak ed be es oe e ae ed 5 1 24 Powering Up and Shutting DOWN a 2s 9 kien ia ee ek bode eae eee e a AR Ae eee 5 122 5 PrimN INE SYSIOM
62. tic E nanoparticles Incubate 10 minutes O Place tube in magnet for 5 minutes Pour off supernatant Positively selected cells remain in tube After two or more rinses collect cells simply by removing tube In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 e mail info stemcellfrance com StemCell Technologies Inc www stemcell com Version 1 1 0 August 2005 Catalog 28940 1 1 2 Robotic System Overview RoboSep is a pipetting robot comprising 4 main mechanical components that are operated under computer control Figure 3 The Robotic Arm The Carousel The Tip Stripping Arm The Hydraulic System Figure 3 RoboSep Instrument Lid Robotic Arm Touch Screen l Carouse Buffer Bottle With these systems acting in concert RoboSep performs the following actions to automate EasySep accurately dispenses magnetic labeling reagents into the cell samples using sterile disposable 1 mL tips mixes the reagents and samples using a sterile disposable 5 mL tip transfers the sample to the magnet for
63. ure only top cap is removed Missed tip stripping action Tip stripping arm not moving Arm is out of position Power down the instrument open the lid and pull the tip stripping arm out Restart RoboSep Tip stripping arm not fully engaging tip head Contact STI for calibration Priming step missing or incomplete No water ejected from tip head during priming step prior to run Connections in hydraulic system are leaking air Make sure the connection between the hydraulic fluid bottle and tubing is seated correctly by disconnecting and reconnecting firmly Hydraulic fluid bottle is empty Refill bottle FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe Tel 33 0 4 76 04 75 30 Fax 33 0 4 76 18 99 63 e mail info stemcellfrance com StemCell Technologies Inc www stemcell com Version 1 1 0 August 2005 Catalog 28940 7 Specifications Capacity Labels and separates up to 4 samples simultaneously Up to 8 x 10 total cells four samples of up to 2 x 109 cells each Sample volume from 250 uL to 8 5 mL for each sample
64. vent shall STI be liable whether in contract or tort including negligence for damages in excess of the purchase price of the Product or for any indirect incidental special or consequential damages of any kind or loss of revenue or profits loss of business or other financial loss arising out of or in connection with the ability or inability to use the Product to the full extent these damages may be disclaimed by law StemCell Technologies Inc makes and or authorizes no other warranty except as is specifically described above No additional warranties may be granted orally or in writing by any representative nor implied by or inferred from the terms above Extended Warranty An optional annual limited warranty extension for up to 2 years can be purchased with the same conditions as above FOR RESEARCH USE ONLY In North America Tel 604 877 0713 Fax 604 877 0704 Toll Free Tel 1 800 667 0322 Toll Free Fax 1 800 567 2899 e mail info stemcell com e mail orders stemcell com In the United Kingdom Tel 44 0 20 7537 7565 Fax 44 0 20 7515 5408 Toll Free within United Kingdom Tel 0800 731 27 14 Fax 0800 731 27 13 e mail info stemcellgb com In Europe i Tel 33 0 4 76 04 75 30 StemCell Technologies Inc Fax 33 0 4 76 18 99 63 www stemcell com e mail info stemcellfrance com Version 1 1 0 August 2005 Catalog 28940 10 Contact Information For customer service technical support or other inform

USER MANUAL

Contents

Download Pdf Manuals

Related Search

Related Contents