Luna-II_user manual_ver1.2.7 (11-10-2014)
Contents
1. 41 4 3 Loading Samples into the Luna Cell Counting Slide 41 4 4 Counting Cells 42 4 5 After Counting Image View 45 4 6 After Counting Histogram amp Gating 47 4 7 After Counting Saving and Printing the Results 50 4 8 After Counting Calculation for Subsequent Experiments 53 3 lyo biosystems www logosbio com Chapter 5 Focusing Option 54 5 1 Autofocusing 54 5 2 Manual Focusing 55 Chapter 6 Review the Previous Results 56 Chapter 7 Maintenance and Troubleshooting 59 7 1 Turn On Off LUNA II 59 7 2 Cleaning 59 7 3 Calibrating the Touchscreen2. 60 7 4 Calibrating LUNA II 60 7 5 Updating the Software 55 7 6 Troubleshooting 61 Chapter 8 Ordering Information 62 Chapter 9 Purchaser Notification 63 9 1 Limited Use Label License 63 9 2 Instrument Warranty 65 Contact Information 2 2 2 2 2 2 2 2 66 A lyo biosystems www logosbio com Safety Information For best results with the LUNA II Automated Cell Counter follow the instructions below in addition to the general precautions for using electrical instruments 11 www logosbio com Be careful to avoid electric shock while operating the instrument Do not touch it and other components with wet hands Do not place it in a humid environment such as an incubator For operating environment see page 9 Trypan blue stain is Known as a hazardous material While handling the solution always wear proper personal protective equipment PPE to avoid exposure Before use make sure
3. 15 2 1 Installation 15 2 2 Start Up Screen 17 2 3 Settings 18 2 4 Calibrating the Touchscreen Touch Calibration 20 2 5 Changing Options for Cell Counting Counting Options 22 2 6 Setting the Date and Time 25 2 7 Calibrating LUNA II Calibrate 26 2 8 Updating the Software Update Software 30 Chapter 3 Protocol Setting 32 3 1 Parameters in the Protocol 32 3 2 Setting the Protocols 34 3 3 Selection of Protocol 38 Chapter 4 Counting Cells 40 4 1 Preparing the Instrument 40 4 2 Sample Preparation
4. and 4X magnification of the image The captured image is also able to be navigated by pressing the screen and moving the finger or stylus pen on the image Two small red boxes in the upper left corner of cell image represent the entire field and field of view respectively The outer box represents the entire field of image And the inner box represents current field of view Upon navigating the image the location of inner box will be changed 45 lyo biosystems www logosbio com Tag in the upper right corner of the image will activate the Tag function The live cells will be marked by green circles and dead cells will be marked by red circles as below Protocol DEFAULT Date 05 Sep 2014 13 33 Live cells Note This Tag function is one of the distinct tools of the LUNA II Automated Cell Counter This function allows the user to review the data immediately to determine the accuracy of the counting without additional manipulation or devices After reviewing the image of cells Tag button can be pressed again to remove the green and red circles from the image 46 lyo biosystems www logosbio com 4 6 After Counting Histogram amp Gating LUNA II provides graphical analyses of the cell counting results By pressing Histogram amp Gating you can view the Histogram amp Gating screen as below Protocol DEFAULT Date 05 Sep 2014 13 33 Min _3_ um Max 60 um amp Cluster
5. www logosbio com In the Review Files users can select File name to review the results and image After selecting the file name the previous cell counting results will be displayed on the right side of screen as below Protocol DEFAULT Date 05 Sep 2014 13 38 eV EEE Total cell 8 16x10e5 cells mL HL 60 05092014 1335 Live cell 6 63x10e5 cells mL Dead cell 1 53x10e5 cells mL L2_no tb Viability 81 2 LINA II test Yw_0409201 9 size 13 8 um Total cell number 176 cells LINA II test YW_04092 Live cell number 143 cells Dead cell number 33 cells Dilution factor 2 luminex bead luna2 LUNA II teat yw 0409201 The captured image with tags can be also reviewed by pressing the small image under the Results section as above The magnification of image can be adjusted by pressing the magnifier on the image 57 biosystems www logosbio com In the Previous Counts function users can overview the list of previous counts with summarized results as below This list can be exported into USB drive as a CSV format The LUNA II Automated Cell Counter can store up to 1 000 previous counts www logosbio com Name Date Total Cell Live Cell Dead Cell HL 60 05092014 13 05 09 2014 13 32 04 09 2014 16 36 04 09 2014 10 47 LINA II test YW_0409201 1 04 09 2014 10 22 LUNA II teat yw 0409201 04 09 2014 10 20 04 09 2014 10 10 8 16E05 176 4 64E03 6 26E0
6. DEFAULT a Medium The selected protocol is marked with white letters and blue back ground in the left panel of screen The value of each parameter is displayed in the right panel of screen 34 lyo biosystems www logosbio com Note The factory setting DEFAULT protocol cannot be modified Protocol DEFAULT Date 05 Sep 2014 10 09 Protocol DEFAULT aut T eo eee ae i Ty Bae DEF AULI protocol canno als New Protocol edited or deleted Please select another protocol RQ 5 50 Medium OK i peme ee Press other protocol or New Protocol and press Load as below A Protocol New Protocol Date 05 Sep 2014 10 09 DEFAULT New Protocol 3 OQ Medium Now LUNA II display following sign on the screen Protocol DEFAULT Date 05 Sep 2014 10 09 New Protocol Selected protocol was loaded 50 Medium OK 35 www logosbio com biosystems Press OK To delete the selected protocol press Delete To edit the selected protocol press Edit ry Protocol New Protocol DEFAULT Date 05 Sep 2014 10 09 New Protocol Medium After pressing Edit the arrows upward and downward will be activated and marked as solid arrows as below amp Protocol New Protocol Date 05 Sep 2014 10 09 DEFAULT New Protocol A A 60 Medium v W Press the arrows to edit the values Then press Save as 36 lyo biosystems www l
7. Start to initiate the Calibration Step 2 Following screen will be displayed with increasing red bar over the time Do not turn off the instrument Protocol DEFAULT Date 04 Sep 2014 14 42 Settings Calibration Step 2 Processing Please do not remove the counting slide nor turn off the instrument during the calibration After completing calibration the instrument will display following sign on the screen Protocol DEFAULT Settings Date 04 Sep 2014 14 43 Calibration is Completed Press Exit for further use Press Exit to move to the Settings screen 28 biosystems iNW logosbio com Now the date of last calibration is changed Compare to the Settings screen in page 26 www logosbio com Settings Last Calibration Calibrated Value Last Update Software Version Last Calibration Calibrated Value 29 Protocol DEFAULT Date 06 Nov 2014 17 30 06 Nov 2014 16 16 0x029D 06 Nov 2014 16 16 1 4 0 06 Nov 2014 17 29 13830 4 2236112 78 8543 2420964 65536 800 480 biosystems 2 8 Updating the Software Update Software Logos Biosystems provides continuous update of software to maintain optimal performance The current version of software can be identified either in the Initializing screen see page 15 or the Settings screen as above The up to date software consisting of single file can be downloaded from Logos Biosystems Website http www log
8. lower and upper limit and two arrow head in the lower part of screen as below Protocol DEFAULT Date 05 Sep 2014 13 34 Min _2_ um Max 41 Jum Se Q z a O O 70 8 90 Cell size um The lower and upper limit can be activated by pressing as above and adjusted by pressing the icons with arrow head The gating function will be helpful to analyze the cells from co culture of cells with distinct sizes of cells The gating function will be also helpful to exclude non cellular particles with distinct sizes from various tissue engineering applications 49 lyo biosystems www logosbio com 4 7 After Counting Saving and Printing the Results The LUNA II Automated Cell Counter provides multiple record options saving the results and or printing the results To save or print the results press Save Print in the Cell Counting Results screen as below Protocol DEFAULT Date 05 Sep 2014 13 32 Next Count 2 Image Total cell concentration 8 16x10e5 cells mL Live cell concentration 6 63x10e5 cells mL Dead cell concentration 1 53x10e5 cells mL llil Ba Viability 81 2 Avg size 13 8 um Total cell number 176 cells Live cell number 4 II Dilution 143 cells Dead cell number 33 cells Dilution factor 2 The Save Print screen will be seen as below In the upper part of screen three items will be displayed in inactive status Items Analyzed image Raw image R
9. the buttons will be inactivated and the bar which is indicating the progress of cell counting will be appeared on the screen as below Staining option With TB Protocol DEFAULT Date 05 Sep 2014 13 32 Counting cells In general the counting takes 10 to 15 seconds LUNA II will display the Cell Counting Results screen as below Total cell concentration Live cell concentration Dead cell concentration Viability Avg size Total cell number Live cell number Dead cell number Dilution factor 44 www logosbio com Protocol DEFAULT Date 05 Sep 2014 13 32 8 16x10e5 cells mL 6 63x10e5 cells mL 1 53x10e5 cells mL 81 2 13 8 um 176 cells 143 cells 33 cells 2 biosystems 4 5 After Counting Image View To view the captured image of cells press Image as below Protocol DEFAULT Date 05 Sep 2014 13 32 Total cell concentration 8 16x10e5 cells mL Live cell concentration 6 63x10e5 cells mL Dead cell concentration 1 53x10e5 cells mL Viability 81 2 Avg size 13 8 um Total cell number 176 cells Live cell number 143 cells Dead cell number 33 cells Dilution factor 2 The Image screen will be appeared as below Protocol DEFAULT Date 05 Sep 2014 13 33 Similar to the Count screen the magnification of the captured image can be changed by pressing the magnifier button on the right of the image The LUNA II Automated Cell Counter provides 1X 2X
10. which are provided in the product package Be sure to check the electrical outlet configuration in your country E USB ports as e__e Power inlet gt e 13 ljos biosystems www logosbio com 1 5 3 Right side view of the LUNA II Automated Cell Counter The right side view of the LUNA II Automated Cell Counter shows a built in printer L40001 model only which allows rapid printing the cell counting results Built in printer 1 5 5 Luna Cell Counting Slide The Luna Cell Counting Slide is a plastic disposable cell counting slide consists of 2 chambers labeled as Aor B that can be used for the same sample as duplicate or for 2 different samples The depth of the counting chamber is 100 um The volume of cells counted is about 0 5 ul almost the same as five 1 mm x 1 mm squares in a standard hemocytometer Sample loading ports Chamber designations Sample chambers A or B respectively 14 lyo biosystems www logosbio com Chapter 2 Setting up 2 1 Installation Upon receiving the product package unpack it carefully and ensure that every component is included and no damage has been occurred Place the LUNA II Automated Cell Counter on a flat and stable surface Insert one end of the power cord into the instrument and plug the other end of the power cord into an electrical outlet after checking the outlet configuration in your local area Turn on the instrument using the po
11. 236112 78 8543 2420964 65536 800 480 22 lyo biosystems www logosbio com In the Counting Options screen current option will be marked as blue V mark as below Counting Options The counting option can be easily changed by pressing un selected option If Without Trypan Blue is chosen the following screen will be displayed Counting Options Without Trypan Blue was selected as a staining option Be sure to check the Dilution Factor at the Protocol menu and use the correct Dilution Factor Press OK to continue 23 www logosbio com lyo biosystems If With Trypan Blue is chosen the following sign will be displayed Counting Options With Trypan Blue 1 1 was selected as a staining option Be sure to check the Dilution Factor at the Protocol menu and use the correct Dilution Factor Press OK to continue The Counting Option is simply changed by pressing OK Otherwise press Cancel to return the previous option Press lt in the upper left corner to move to the Settings screen 24 lyo biosystems www logosbio com 2 6 Setting the Date and Time The LUNA II Automated Cell Counter provides current date and time for record keeping Users may want to change the date and time since factory setting is adjusted to the local time of Korean Once the date and time are set no additional setting is required for routi
12. 5 Focusing Option The LUNA II Automated Cell Counter has been integrated with a novel focusing mechanism based on the liquid lens technology Unlike traditional autofocusing based on mechanical moving of lens in vertical axis the liquid lens technology enables autofocusing without mechanical moving With this advance the LUNA II Automated Cell Counter provides two focusing options Autofocusing and manual focusing 5 1 Autofocusing With autofocusing function the LUNA II Automated Cell Counter achieves true automation of cell counting Users count cells by simply insert the cell counting slide and pressing Count without tedious focusing process In addition autofocusing function eliminates potential human error Autofocusing function can be activated by simply pressing Autofocused Counting in the lower left corner of the Count screen When activated the blue circle will be displayed on the right side of Autofocused Counting as below Staining option Protocol DEFAULT With TB Date 05 Sep 2014 16 10 As shown above the Luna Standard Beads are out of focus However users can count the beads without tedious focusing 54 lyo biosystems www logosbio com After pressing Count users can obtain autofocused image as below Protocol DEFAULT Date 05 Sep 2014 16 11 5 2 Manual Focusing Although the LUNA II Automated Cell Counter provides autofocusing function users may want to focu
13. 5 135 6 63E05 143 7 10E05 153 0 00E00 0 58 6 63E05 143 0 00E00 4 64E03 1 0 00E00 0 9 27E03 2 Protocol DEFAULT Date 05 Sep 2014 13 36 4 vib AVO Protocol 1 53E05 33 4 64E03 6 22E05 134 6 63E05 143 7 00E05 151 81 2 13 8 DEFAULT DEFAULT DEFAULT DEFAULT DEFAULT DEFAULT biosystems Chapter 7 Maintenance and Troubleshooting 7 1 Turn On Off LUNA II To turn on LUNA II press the POWER button in front of the instrument The Power icon in the Start Up screen can be used to turn off the instrument See page 17 Alternatively the instrument is turned off by pressing the POWER button in front of the instrument for 5 seconds Since LUNA II provides standby mode it is not necessary to turn off the instrument in general laboratory use Standby mode will be activated after 10 min of inactivity In the standby mode the touchscreen will blackout By simply pressing the touchscreen or the POWER button the LUNA II will be ready to use with displaying the last screen 7 2 Cleaning Generally the LUNA II Automated Cell Counter does not require regular maintenance for appropriate operation However if the instrument is used for long periods of time and continuously it may need to be cleaned or decontaminated to remove any dirt or dust on the surface of the instrument Be sure to turn off the LUNA II Automated Cell Counter a
14. A II see section 2 7 7 5 Updating the Software see section 2 8 IMPORTANT The re calibration must be done after the firmware update Please see Section 2 3 4 60 lyo biosystems www logosbio com 7 6 Troubleshooting Clumped cells Concentration of cells Insertion of counting slide Inaccurate cell Sample loading count Malfunction of optical components Damage or contamination of counting slide Incompatible USB drive Data transfer and saving Too many files in the USB drive Errors during updating and Freezing during calibration calibrating the instrument www logosbio com Make sure that cells are not clumped The more single cells the better counting results For best results the concentration of cells should be 5 x 104 1 x 10 cells ml If needed dilute or concentrate the cells within this range Ensure that the counting slide is inserted properly into the instrument If the counting slide is over or under loaded with the sample it may affect counting results The optimal volume of sample is 10 12 ul of cell suspension Any of the optical components may be damaged Or the objective lens may be dirty due to dust spilled samples or unknown causes Please contact your local supplier Make sure that the counting area of the slide is transparent before loading the sample Wear gloves while handling the slide Use the USB drive supplied with the instrument O
15. IF format Optimized for LUNA II only LUNA II software www logosbio com PDF report Approx 10 without autofocusing or 15 with autofocusing seconds at 1 x 10 cell ml concentration Processing time may vary by cell type and concentration 1 4 2 Luna Cell Counting Slide specifications Material Dimensions WxDxH Chamber Depth Chamber Volume Polystyrene 25 X 75x 2 4 mm 100 um 10 ul Note One Luna USB Drive 4 Gigabytes and 0 4 Trypan Blue Stain solution 2 x 1 ml are included in the starter package www logosbio com lyo biosystems 1 5 Product Description 1 5 1 Front view of the LUNA II Automated Cell Counter The front view of the LUNA II Automated Cell Counter shows various parts as shown below e Wide touchscreen located in the upper front of the instruments contains buttons for all functions needed to operate the instrument and displays data acquired e Power button is used to turn on the instrument e Counting slide port is used to insert the Luna Cell Counting Slide containing sample to analyze e The front USB port provides easy to access data retrieving to transfer Touchscreen Power button Counting slide port USB port 1 5 2 Rear view of the LUNA II Automated Cell Counter The rear view of the LUNA II Automated Cell Counter shows two additional USB ports and a power inlet to connect the instrument to an electrical outlet with the power cord and plug
16. LUNA H Automated Cell Counter Catalog No L40001 L4000 Document No L44001 Revision Date November 07 2014 SW Version 1 2 7 ljo biosystems www logosbio com IMPORTANT The LUNA II Automated Cell Counter is a Laboratory Electrical Instrument for Scientific Research Use Only BUT NOT A MEDICAL or THERAPEUTIC or IN VITRO DIAGNOTICS DEVICE lyo biosystems www logosbio com Table of Contents Safety Information 5 General Guidelines 7 Environment Conditions 8 Chapter 1 Introduction 9 1 1 Product Overview 9 1 2 Key Features 10 1 3 Product Contents 11 1 4 Product Specifications 12 1 5 Product Description 13 Chapter 2 Setting up
17. ULT With TB Date 05 Sep 2014 12 38 In the Count screen users can identify the Staining option and selected protocol in the upper panel of screen To change the Staining option see Section 2 5 Changing Options for Cell Counting To change the Protocol see Section 3 3 Selection of Protocol Users can also identify the date and time in the upper right corner of screen To change the date and time see Section 2 6 Setting the Date and Time Now LUNA II is ready to count 40 lyo biosystems www logosbio com 4 2 Sample Preparation 4 2 1 Materials required Cell suspension Luna Cell Counting Slides Trypan Blue Stain 0 4 Luna USB drive or equivalent 4 2 2 Prepare cell suspension according to standard cell biology procedure Please avoid clumping of cells for accurate counting 4 2 3 Mix 10 ul of the cell suspension with 10 ul of Trypan Blue Stain in an appropriate tube by gentle pipetting up and down 4 3 Loading Samples into the Luna Cell Counting Slide Hold the edge of the slide and load 10 12 ul of the mixed cell sample into the sample loading port of one chamber of the counting slide Alternatively the slide can be placed on a clean surface during sample loading For easy and accurate loading tilt the pipette 45 60 degrees as shown below Note Be careful not to over load or under load the sample into the chamber 41 4 4 Counting Cells 4 4 1 Insert the sample loaded sli
18. WEEE in an ecologically friendly manner Follow waste ordinances of your region for proper disposal provisions The CE mark indicates that this instrument conforms to all applicable European Community provisions for which this marking is required Users must be aware of C and follow the conditions described in this manual for operating the instrument The protection provided by the instrument may be impaired if the instrument is used in a manner not specified by this manual D Protective earth Ground ljos biosystems www logosbio com General Guidelines for Using the LUNA II Automated Cell Counter In order to achieve the best results with the LUNA II Automated Cell Counter follow the instructions below carefully 1 The instrument must be operated in compliance with the operating environment described on page 9 In particular the temperature and humidity conditions are important 2 Samples must be handled in an appropriate way depending on user s requirements 3 Hold the Luna Cell Counting Slide by the edges to avoid touching its optical surface Make sure that no damage or contamination occurs on the optical surfaces of the slide 4 After mixing the cell sample with trypan blue stain perform cell counting as soon as possible within 3 minutes for accurate cell viability measurement If needed count your sample at least 2 times duplicate readings and take an average 5 Since the LUNA II Aut
19. WETS Cell number 70 80 90 Cell size um On this screen users can review the distribution of cells according to their sizes Green bars represent the live cells and red bars represent dead cells The histogram will change by pressing two buttons on the right side of screen When pressing Total on the icon will be changes as Live on and the histogram will display the distribution of live cells without dead cells as below Protocol DEFAULT Date 05 Sep 2014 13 33 Min _ 3 _ um Max 60 um eS Cell number Cell size um 47 lyo biosystems www logosbio com When pressing Live on the icon will be changes as Dead on and the histogram will display the distribution of dead cells without live cells as below Protocol DEFAULT Date 05 Sep 2014 13 33 Cell number Cell size um In addition when pressing Cell number on the left side of histogram the LUNA II Automated Cell Counter also display the cell concentration The LUNA II Automated Cell Counter also provides the distribution of clusters as below after pressing Cluster Map off Protocol DEFAULT Date 05 Sep 2014 13 33 Min _3_ um Max 60 um eS 1 cell 2 cell 3 cell 4 cell 5 cell 6 cell 7 cell gt 7 cell foo Cell cluster 48 lyo biosystems www logosbio com In addition the LUNA II Automated Cell Counter provides gating function Gating function consists of four components the
20. aterials made using this product or its components to a third party for Commercial Purposes Commercial Purposes mean any and all uses of this product and its components by a party for monetary or other consideration including but not limited to a product manufacture b providing a service information or data c therapeutic diagnostic or prophylactic purposes or d resale of this product or its components whether or not such product and its components are resold for use in research Logos Biosystems Inc Company will not claim any consideration against the purchaser of infringement of patents owned or controlled by the Company which cover the product based on the manufacture use or sale of a therapeutic clinical diagnostic vaccine or prophylactic product developed in research by the purchaser in which this product or its components was employed provided that neither this product nor any of its components was used in the manufacture of such product For any use other than this limited use label license of research use only please contact the Company or e mail to info logosbio com for more information 64 lyo biosystems www logosbio com 9 2 Instrument Warranty Logos Biosystems Inc Company warrants to the original purchaser Purchaser that the instrument Instrument if properly used and installed will be free from defects in materials and workmanship and will conform to the p
21. cell counting With Trypan Blue or Without Trypan Blue This function provides inter change of these options Option Description With Trypan Blue This option can be used for regular bright field counting when cell samples are mixed with 0 4 trypan blue stain in a 1 1 ratio This option can generate cell viability data For cell counting with this option the dilution factor in the Protocol should be set to value 2 Without Trypan Blue When samples do not contain trypan blue stain turn on this option and follow the directions in the message boxes For cell counting with this option please make sure that the dilution factor in the Protocol should be set to value 1 The cell counting parameters of LUNA II are optimized with the use of trypan blue Low contrast from no use of trypan blue stain may cause abnormal results The Dilution Factor in the Protocol is not changed automatically After changing the Counting Options the Dilution Factor should be changed manually Improper use of the Dilution Factor causes incorrect calculation of cell concentrations On the Settings screen press Counting Options Protocol DEFAULT Date 06 Nov 2014 17 30 Settings Date Time Calibration 31 Jul 2014 17 05 0x029D Last Update 06 Nov 2014 16 16 Update Software Software Version 1 4 0 re Touch Last Calibration 06 Nov 2014 17 29 Calibration Calibrated Value 13830 4 2
22. de into the counting slide port of LUNA II ensuring that the loaded chamber is inserted into the instrument LUNA II analyzes only the inserted chamber upon counting Note After inserting the slide LUNA II only reads the inserted chamber Note Make sure that the counting slide is not inserted upside down Now users can see the cells on the screen as below Staining option Protocol DEFAULT With TB Date 05 Sep 2014 13 32 Note If users cannot see the cells the cell counting slide may not be inserted correctly The magnification of cells can be changed by pressing the magnifier as below Staining option Protocol DEFAULT With TB BEIC 05 Sep 2014 13 32 The LUNA II Automated Cell Counter provides 1X 2X and 4X images as follows ljo biosystems 42 www logosbio com Staining option Protocol DEFAULT With TB Date 05 Sep 2014 13 32 Staining option Protocol DEFAULT With TB Date 05 Sep 2014 13 32 The field of cell image can be navigated by pressing the screen and moving the finger or stylus pen on the image Two small red boxes in the upper left corner of cell image represent the entire field and field of view respectively The outer box represents the entire field of image And the inner box represents current field of view Upon navigating the image the location of inner box will be changed 43 www logosbio com ljo biosystems 4 4 2 Press Count to start counting Now all of
23. eport www logosbio com iv ole aa iw Raw Image iv Report POOE Bi Mam E Description The image of cells with tags of live and dead cells The image of cells without tag The PDF report containing counting results and histograms 50 lyo biosystems These items can be activated by pressing and displayed with blue V marks as below PXolom B Wam E Put appropriate name such as the name of cells using keyboard on the screen as above Optionally users can put the date and time by simply clicking Add Date Time in the lower right corner of screen T m jl TER me Raw Image iw Report POOR Bi Wa E Now users can save the selected data by pressing Save in the lower right corner of screen IMPORTANT Remember that the Raw Image should be turned on to save the Raw Image which should be provided to your distributor or Logos Biosystems to get the best technical support 51 lyo biosystems www logosbio com The LUNA II Automated Cell Counter provides additional option to report the results Users easily print the Cell Counting Results by simply pressing Print in the upper right corner of screen The printed report will be displayed as below Cell Count Report File name HL 60 05092014 1 3350605 Date 05 Sep 2014 13 35 Cell count results Total 8 16x10e5 cells mL Live 6 63x10e5 cells mL Dead 1 53x10e5 cells mL Viability 81 2 Avg si
24. ersion 1 4 0 Last Calibration 06 Nov 2014 17 29 Calibrated Value 13830 4 2236112 78 8543 2420964 65536 800 480 Now users can see the following screen If users want to calibrate the touchscreen press OK If not press Cancel Protocol DEFAULT Date 04 Sep 2014 14 12 Settings if your instrument is not responding properly to your taps you may need to recalibrate your sceen Recalibration involves tapping the center of a target If you miss the center keep the stylus on the screen slide it over the target s center and then lift the stylus To Start tap Recalibrate 20 www logosbio com biosystems Now small cross mark will be appeared in the upper left corner in the grey screen as below Press the cross mark then the cross mark will move to the upper right corner as below Press the cross mark again The mark will move to lower right corner of screen Press it again The cross mark will further move to lower left corner and center of screen after each touch After touch the cross mark in the center the following sign will be displayed in the center of screen Touch calibration completed After a few seconds the Settings screen will be re appeared spontaneously Press the Home image to go to the Start Up screen 21 lyo biosystems www logosbio com 2 5 Changing Options for Cell Counting Counting Options The LUNA II Automated Cell Counter provides two options for
25. essary the travel cost of a service engineer 66 ljos biosystems www logosbio com Contact Information For more information or technical support please call write fax or email Our regional suppliers are listed on our web page www logosbio com Logos Biosystems Inc Address Suite 930 Doosan Venturedigm 126 1 Pyungchon Dong Dongan Gu Anyang City Gyunggi Do Republic of Korea Zip code 431 755 Tel 82 31 478 4185 Fax 82 31 478 4184 E mail sales logosbio com 67 ljos biosystems www logosbio com
26. gosbio com Chapter 8 Ordering Information The following products can be ordered from your regional supplier or the website www logosbio com Cat L40001 L40002 L12001 L12002 L12003 T13001 B13001 U10004 P12001 Product LUNA II Automated Cell Counter with printer LUNA II Automated Cell Counter without printer Luna Cell Counting Slides 50 slides 100 counts Luna Cell Counting Slides 500 slides 1 000 counts Luna Cell Counting Slides 1 000 slides 2 000 counts Trypan blue stain 0 4 for use with LUNA II Luna Standard Bead Luna USB Drive 4 Gigabytes LUNA II Printer Paper 10 pk min 700 prints 63 www logosbio com Size each each 1 box 10 boxes 20 boxes 2x1ml 2x1ml each each lyo biosystems Chapter 9 Purchaser Notification 9 1 Limited Use Label License Research Use Only The purchaser of this product should use this product only for research for the sole benefit of the purchaser By use of this product the purchaser agrees to be bounded by the terms of this limited use statement whether the purchaser is a for profit or a not for profit entity If the purchaser is not willing to accept the conditions of this limited use statement and this product is unused the Company will accept return of the product with a full refund The purchaser cannot re sell or otherwise transfer a this product b its components or c m
27. he Start Up screen as below review protocol settings Then the following Settings screen will be displayed Protocol DEFAULT Settings Date 06 Nov 2014 17 30 Last Calibration 31 Jul 2014 17 05 Calibrated Value 0x029D Last Update 06 Nov 2014 16 16 Software Version 1 4 0 Last Calibration 06 Nov 2014 17 29 Calibrated Value 13830 4 2236112 78 8543 2420964 65536 800 480 In the Settings screen users can see the followings www logosbio com Home image by Pressing Home image you can go to the Start Up screen Current protocol and date in the upper right corner Date and values of last calibration Date and software version of latest software update 18 lyo biosystems The Settings menu allows you to perform the followings Calibrate the touchscreen by pressing Touch Calibration Change options for cell counting by pressing Counting Options Set the date and or time by pressing Date Time Re calibrate the instrument by pressing Calibrate Update the software by pressing Update Software www logosbio com ljos biosystems 2 4 Calibrating the Touchscreen Touch Calibration This function can be used when the response of touchscreen is abnormal On the Settings screen press Touch Calibration as below Protocol DEFAULT Settings Date 06 Nov 2014 17 30 Last Calibration 31 Jul 2014 17 05 Calibrated Value 0x029D Last Update 06 Nov 2014 16 16 Software V
28. ill be helpful for optimal detection of specific type of cells Roundness Roundness refers to the roundness of the objects in the image Since the shape of cells may vary and not complete sphere adjusting Roundness enables optimal detection of cells The instrument with higher Roundness counts objects with more roundness as cells and excludes objects with less roundness for counting as cells LUNA II with lower Roundness will be suitable to count cells with irregular shape since it detects objects with less roundness as cells 32 lyo biosystems www logosbio com Min amp Max Cell Size Average size of cells also may vary from cell type to cell type With this parameter users can optimize the instrument to efficiently detect their own cells The value can be adjusted with 1 um of increase per step Declustering Level Cultured mammalian cells may form clumps during culture or handling Declustering function of LUNA II provides efficient detection of clumped cells LUNA II provides 4 alternates None Low Medium and High This function is helpful to count sticky cells or rod shaped spores High level declustering takes more time to analyze 33 ljos biosystems www logosbio com 3 2 Setting the Protocols In the Start Up screen press protocol as below review protocol settings In the Protocol screen users can identify all the parameters as below A Protocol DEFAULT Date 05 Sep 2014 10 08
29. nd disconnect the power cable before cleaning or performing any other maintenance Ensure that water and other solutions do not enter any part of the instrument during cleaning 7 2 1 Cleaning the surface With a soft and damp cloth wipe the surface of the instrument Use some distilled water or alcohol for dampening the cloth After cleaning immediately dry the cell counter with a dry cloth Do not wet the instrument by pouring or spraying water or other liquids directly on the instrument In particular power related parts should never become wet in order to avoid electrical shock or damage 7 2 2 Cleaning the touchscreen Gently wipe off the touchscreen with a soft cloth lightly moistened with an authorized LCD cleansing detergent Since excessive force or pressure on the touchscreen can cause damage be gentle and cautious during cleaning Wipe the touch screen dry immediately 59 lyo biosystems www logosbio com 7 2 3 Decontaminating with alcohol When the instrument needs to be decontaminated use a soft cloth lightly moistened with 70 alcohol to wipe the outer surface Never pour or spray alcohol or any other solution directly onto the instrument this may cause severe damage to the instrument or give an electric shock to users Note do not use an abrasive solution or a bleach solution that can cause scratches on the outer surface or the touchscreen 7 3 Calibrating the Touchscreen see section 2 4 7 4 Calibrating LUN
30. ne laboratory use On the Settings screen press Date Time as below Setti Protocol DEFAULT ettings Date 06 Nov 2014 17 30 Last Calibration 31 Jul 2014 17 05 Calibrated Value 0x029D Last Update 06 Nov 2014 16 16 Software Version 1 4 0 Last Calibration 06 Nov 2014 17 29 Calibrated Value 13830 4 2236112 78 8543 2420964 65536 800 480 The following screen will be displayed Date Time YYYY Hour 4 09 ms 40 Press desired field to erase the current number Put number by pressing numbers on the right panel and press Apply to save changes Press lt in the upper left corner to move to the Settings screen 25 lyo biosystems www logosbio com 2 Calibrating LUNA II Calibrate Background calibration is a prerequisite for successful detection of cells The LUNA II Automated Cell Counter provides easy to use automatic self calibration of background On the Settings screen press Calibrate as below Setti Protocol DEFAULT ettings Date 06 Nov 2014 17 30 Last Calibration 31 Jul 2014 17 05 Calibrated Value 0x029D Last Update 06 Nov 2014 16 16 Software Version 1 4 0 Last Calibration 06 Nov 2014 17 29 Calibrated Value 13830 4 2236112 78 8543 2420964 65536 800 480 The following screen will be displayed S tti Protocol DEFAULT etungs Date 04 Sep 2014 14 40 Calibration Step 1 Remove the counting slide from the instrument and press sta
31. nt is correct at its publication However the Company makes no warranty of any kind regarding the contents of any publications or documentation as unintended or unexpected errors including occasional typographies or other kinds are inevitable In addition the Company reserves the right to make any changes necessary without notice as part of ongoing product development If you discover an error in any of our publications please report it to your local supplier or the Company 65 lyo biosystems www logosbio com The Company shall have no responsibility or liability for any special incidental indirect or consequential loss or damage resulting from the use or malfunction of the Instrument This limited warranty is sole and exclusive The Company makes no other representations or warranties of any kind either express or implied including for merchantability or fitness for a particular purpose with regards to this Instrument To obtain service during the Warranty Period contact your local supplier or the Company s Technical Support team OUT OF WARRANTY SERVICE Please contact your local supplier or the Company s Technical Support team in order to obtain out of warranty service If necessary repair service will be charged for replacement parts and labor hours incurred to repair the Instrument In addition the Purchaser is responsible for the cost of shipping the Instrument to and from the service facility and if nec
32. ocol Documentation www logosbio com Description Compact size with light weight saves space and is suitable for either laboratory table or biosafety cabinet With sophisticated optical components and counting algorithm LUNA II provides optimized and reproducible results every time Advanced liquid lens technology provides fast autofocusing without mechanical moving Autofocusing enables reliable cell counting and excludes human error The intuitive user interface based on a touch screen enables simple and easy operation Results for most cell lines are available within 10 without autofocusing or 15 with autofocusing seconds after pressing the Count An integrated thermal printer provides concise Cell Count Report facilitating record keeping LUNA II adopts an innovative counting slide made with T BOND technology without using hazardous organic solvents Measurements can be made for cells at concentrations ranging from 5 x 104 to 1 x 10 cells ml and for cells within the 3 60 um diameter range No external calculator is needed to calculate subsequent dilution Just plug in and it is ready for use with virtually no maintenance costs The captured image of cells can be downloaded onto a USB drive in TIFF Tag Image File Format for review or record keeping Different protocols up to 300 protocols can be saved with personalized parameters LUNA II provides a PDF Portable Document Format rep
33. of trypan blue stain Second load the cell suspension into the Luna Cell Counting Slide Third insert the slide into the slide port of the instrument and chose Autofocused Counting or adjust the focus by clicking arrows to get an appropriate cell image Last press the Count button and then the results of cell count and viability will be displayed on the screen The counting image can be downloaded onto a USB drive in TIF format for review and or record keeping The LUNA II Automated Cell Counter provides key data as below Number of live and dead cells ml Number of total cells ml Viability percentage live cells to total cells Cell images showing live cells as green circles and dead cells as red circles Histograms of cell size distributions The Luna Cell Counting Slide is disposable and specifically designed for the LUNA II Automated Cell Counter Each counting slide has 2 chambers labeled as A and B respectively so that one slide can be used for the same sample reading in duplicate or for 2 different samples if preferred lyo biosystems www logosbio com 1 2 Key Features Key features Small footprint Accuracy amp precision Autofocusing Easy to operate user interface Shortest time to results Built in printer Optional Innovative counting slide Cell concentration amp viability range Easy to use calculator Setup amp maintenance Counting image acquisition Individual prot
34. ogosbio com Now users can see the Save as screen as below if Using keyboard in the screen put the desired name and press Save in the lower right corner if Now new protocol named James is displayed in the Protocol panel of Protocol screen EA Protocol New Protocol Date 05 Sep 2014 10 10 3 lyo biosystems www logosbio com 3 3 Selection of Protocol To change protocol to be used press the protocol button in the Set Up screen review protocol settings Select desired protocol by pressing the protocol name and press Load to apply selected protocol to use A Protocol New Protocol Date 05 Sep 2014 10 10 Medium 38 lyo biosystems www logosbio com LUNA II will display following sign on the screen fi Protocol DEFAULT Date 05 Sep 2014 10 09 DEFAULT New Protocol Selected protocol was loaded 0 Medium Now the instrument is ready to count cells with selected protocol IMPORTANT Selecting the protocol name does not mean changing protocol to be used To use selected protocol users should press Load 39 ljo biosystems vww logosbio com Chapter 4 Counting Cells 4 1 Preparing the Instrument Turn on or activate the LUNA II Automated Cell Counter See Section 7 1 Press count on the Start Up screen o5 review protocol settings LUNA II will display the Count screen Staining option Protocol DEFA
35. omated Cell Counter is calibrated before shipping you do not need to re calibrate before use However if re calibration is needed please refer to Section 5 3 Calibrating the Counter 6 Do not touch trypan blue solution with bare hands as it is a hazardous chemical After using the counting slides dispose of it as hazardous wastes Do not reuse the slides lyo biosystems www logosbio com Environment Conditions Operating Power Frequency Electrical Input Installation Site Operating Temperature Maximum Relative Humidity Altitude Pollution Degree www logosbio com 100 240 VAC 1 2A 50 60 Hz 12 VDC 3 3A Indoor use only 10 35 C 20 80 lt 2 000 m 2 ljos biosystems Chapter 1 Introduction 1 1 Product Overview The LUNA II Automated Cell Counter is a small fast and affordable image based cell counting device that automatically counts various kinds of cells for research purpose The LUNA II Automated Cell Counter helps measure the number as well as viability of cells live dead total cells with sophisticated optical components and advanced image analysis algorithms Due to several innovations introduced by Logos Biosystems LUNA II provides a state of the art cell counting device and eliminates the tedium and subjectivity of manual cell counting The LUNA II Automated Cell Counter can be used in a very simple procedure First mix 10 ul of the cell sample with 10 ul
36. ort which includes File name Date Cell count results Cell images and histograms 10 lyo biosystems 1 3 Product Contents The product package of the LUNA II Automated Cell Counter contains the following components Component Quantity LUNA II Automated Cell Counter with or without printer Power cord including an adapter 1 Luna Cell Counting Slides 1 box 50 slides for 100 counts Trypan Blue Stain 0 4 2x1ml Luna USB drive 1 After receiving the product package please immediately unpack it and check the components listed above to ensure that all parts are included and no damage has been occurred during shipping The warranty does not cover damage that may occur during shipping and handling Any damage claims must be filed with the carrier Note The LUNA II Automated Cell Counter is only for research purpose not for human or animal therapeutic or diagnostic use 11 lyo biosystems www logosbio com 1 4 Product Specifications 1 4 1 LUNA II Automated Cell Counter specifications Instrument Type Dimensions WxDxH Weight Cell Concentration Range Cell Diameter Range Cell Viability Range Image Resolution Image Type Software Documentation Processing Time Benchtop cell counter 16 x 18 x 28 cm 6 3 X 7 0 X 11 0 inch 1 6 kg 3 5 Ib without the external power adaptor 5 x 104 1 x 10 cells ml 3 60 um optimal 8 30 um 0 100 5 mega pixels 5 MP T
37. osbio com This software should be saved into the root directory of the USB drive Before starting download the new software into the Luna USB drive and connect the USB drive to the USB port of the instrument Press Update Software to start Setti Protocol DEFAULT ettings Date 06 Nov 2014 17 30 Last Calibration 06 Nov 2014 16 16 Calibrated Value 0x029D Last Update 06 Nov 2014 16 16 Software Version 1 4 0 l l Last Calibration 06 Nov 2014 17 29 Calibrated Value 13830 4 2236112 78 8543 2420964 65536 800 480 The following sign will be displayed on the touchscreen Protocol DEFAULT Ooa Settings Date 04 Sep 2014 14 43 Please insert USB drive containing the latest software version 30 lyo biosystems www logosbio com Press Start Software update will be proceeded automatically Do not turn off the instrument during update After finishing the current version of software and the date of last update will be changed automatically IMPORTANT The re calibration must be done after the firmware update Please see Section 2 7 31 ljos biosystems www logosbio com Chapter 3 Protocol Setting The LUNA II Automated Cell Counter provides a basic DEFAULT protocol which can be used for most common cell lines In addition LUNA II also provides tools to setup individual protocols All the parameters in the protocol can be modified by users and saved as alternative pro
38. r make sure that your USB drive is compatible with the instrument The version of the USB drive must be 2 0 Some types of USB drives are not detected or compatible with the instrument When there are too many saved files on the USB drive reading and writing by the counter may slow down Generally re calibration takes several minutes However it may take more time depending on the extent of background adjustment If the calibration takes more than 10 minutes reset the system by turning off and on using the power button located in front of the instrument Please contact service engineer if the calibration fails repeatedly 61 lyo biosystems Use the USB drive supplied with the instrument Or make sure that your USB drive is compatible Incompatible USB drive with the instrument The version of the USB drive must be 2 0 Some types of USB drives are not detected or compatible with the instrument More than one software Delete software with previous versions from the version USB drive before downloading new software First make sure that the USB drive works well and is compatible with the instrument Second download the file again onto the USB drive The Incorrectly saved or file should be located in the root directory Third damaged software ensure that the USB drive is inserted correctly Last try the update again If the problem continues contact your local supplier 62 lyo biosystems www lo
39. roduct specifications for a period of one 1 year Warranty Period from the date of purchase If the Instrument under this limited warranty fails during the Warranty Period the Company at its sole responsibility will 1 within and up to 30 calendar days of purchase refund the purchase price of the Instrument to the Purchaser if the Instrument is in original conditions or 2 after 30 calendar days of purchase only replace or repair the Instrument for up to the Warranty Period without issuing a credit In no event shall the Company accept any returned instrument including its components that might have been used or contaminated in some labs including but not limited to HIV or other infectious disease or blood handling labs This limited warranty does not cover refund replacement and repair incurred by accident abuse misuse neglect unauthorized repair or modification of the Instrument This limited warranty will be invalid if the Instrument is disassembled or repaired by the Purchaser In case that the Company decides to repair the Instrument not to replace this limited warranty includes replacement parts and labor for the Instrument This limited warranty does not include shipment of the Instrument to and from service location or travel cost of service engineer the costs of which shall be borne by the Purchaser Every effort has been made to ensure that all the information contained in this docume
40. rt _ A a a For the Calibration Step 1 the counting slide port should be emptied As directed on the screen remove the counting slide from the counting slide port of the instrument Press Start 26 lyo biosystems www logosbio com Following screen will be displayed with increasing red bar over the time S iti Protocol DEFAULT etungs Date 04 Sep 2014 14 40 Calibration Processing Please do not turn off the instrument during the calibration The Calibration Step 1 will take a few seconds Do not turn off the instrument During the process prepare diluted trypan blue solution by mixing 0 4 trypan blue stain with equal volume of either distilled water phosphate buffered saline PBS or plain medium Put 10 ul of diluted trypan blue solution into the chamber of new cell counting slide see Section 4 3 After completion of the Calibration Step 1 following screen will be displayed S tti Protocol DEFAULT ettings Date 04 Sep 2014 14 41 Calibration Step 2 Please insert a counting chamber containing 1 1 diluted trypan blue 0 2 concentration ED D D Now place the cell counting slide containing diluted trypan blue solution into the counting slide port Be sure the chamber containing the trypan blue solution to be placed inside the instrument Do not turn the cell counting slide upside down 27 ljo biosystems www logosbio com Press
41. s the image manually with various purposes The LUNA II Automated Cell Counter also provides manual focusing function Users can adjust the focus manually by simply clicking the arrow heads up or down as below with either activating or deactivating autofocus function Staining option Protocol DEFAULT With TB Date 05 Sep 2014 16 10 qM Ney Note If needed users can count the cells with deactivating autofocus function after manual focusing 55 lyo biosystems www logosbio com Chapter 6 Review the Previous Results The LUNA II Automated Cell Counter provides stand alone review function for previous results Users can easily review the previous results without additional devices To review the previous results the USB drive that contains the previous results should be connected to the USB port of the LUNA II Automated Cell Counter Users can select the Review function by clicking review in the Start Up screen review protocol settings The Review function provides two options Review Files and Previous Counts as below Protocol DEFAULT Date 05 Sep 2014 13 36 HL 60 05092014 1335 Total cell HL 60 05092014 1335 Hve cel Dead cell L2 notb Viability LINA II test Yw_0409201 9 size Total cell number LINA II test YW_04092 ove Live cell number Dead cell number Dilution factor LUNA II teat yw 0409201 luminex bead luna2 56 lyo biosystems
42. that the input voltage is compatible with the instrument s power supply voltage For optimal operation place the instrument on a flat surface and avoid any vibration Turn on the instrument only after connecting both ends of its power cord to the wall outlet as well as the instrument Always turn off the instrument before disconnecting the power cord and or moving the instrument Ensure that the power cord is firmly plugged into the power inlet the wall outlet and AC adapter When the instrument is operating for a long time its temperature can become too high Please be careful that the instrument s temperature does not become too high during long and continuous operation times When operating leave enough space around the instrument so there is enough room for air circulation and cooling Do not disassemble the instrument in any event If the instrument is out of order or dropped or broken please contact a service person Disassembling the instrument invalidates its warranty Use only authorized components adaptor power cord and USB drive If the instrument emits smoke disconnect the power cord immediately from the wall outlet and contact a service person Used counting slides must be disposed as biohazard wastes lyo biosystems lt Symbols used in this User Manual gt The WEEE Waste Electrical and Electronic Equipment symbol indicates that users of this instrument have the responsibility of returning and disposing of
43. tocol Users can save up to 300 protocols These functions provide personalized protocol for optimal use 3 1 Parameters in the Protocol In the protocol menu the LUNA II Automated Cell Counter provides following parameters Parameter Range DEFAULT Dilution Factor 1 100 2 Noise Reduction 0 10 5 Roundness 0 100 60 Min Cell Size 3 59 um 3 Max Cell Size 4 60 um 60 Declustering Level None Low Medium High Medium Dilution Factor The value for dilution factor in the DEFAULT protocol is preset as 2 for With Trypan Blue However users can modify this value according to the dilution of the original sample The Dilution Factor can be adjusted either by a scale of 1 or a scale of 10 between 2 to 10 and 10 100 respectively The Dilution Factor is used to automatically calculate the concentration of cells in original sample from the cell counting result Adjusting Dilution Factor will be helpful for users handling high density cells such as fermented CHO cells In such cases serial dilutions and repeated counting will be needed with appropriate Dilution Factor Noise Reduction Noise Reduction means the decrease of the background for counting With higher Noise Reduction the instrument dose not detect faint signals of weakly stained objects With lower Noise Reduction the instrument can detect objects with faint signals Since staining intensity of cells with trypan blue may vary from cell to cell adjusting Noise Reduction w
44. wer button located on the front of the instrument Initializing screen after company logo will be displayed a few seconds as below Company logo LUNA II Automated Cell Counter ver 1 0 9 21 MAY 2014 Initializing screen of the LUNA II Automated Cell Counter In the initializing screen the current version of LUNA II software can be identified in the right lower corner Initializing screen will be followed by LOADING sign as below 15 ljos biosystems www logosbio com LOADING sign of the LUNA II Automated Cell Counter The Start Up screen will be displayed on the touchscreen as shown below in 2 2 See Section 7 1 for Turn on and Turn off the instrument 16 lyo biosystems www logosbio com 2 2 Start Up Screen The Start Up screen will show 5 compartments Power icon count review protocol and settings Power review protocol settings Main menu The Power icon in the Start Up screen can be used to turn off the instrument See Section 7 1 for further information to turn on off the instrument 17 lyo biosystems www logosbio com 2 3 Settings Generally there is no need to change the settings of the instrument since they are preset at the time of manufacture If users need to reset the date time or other options users can adjust or change the options parameters in the Settings menu described below Press settings located on the right of t
45. ze 13 8 um Total 176 cells Live 143 cells Dead 33 cells Dil Factor 2 Protocol Protocol name DEFAULT Noise reduction 5 Roundness 60 Min cell size 3 Max cell size 60 Size gating 3 60 um 52 ljos biosystems www logosbio com 4 8 After Counting Calculation for Subsequent Experiments The LUNA II Automated Cell Counter provides a built in dilution calculator Users easily calculate the amount of cell suspension to dilute for subsequent experiments Press Dilution to activate the Dilution Calculator as below Protocol DEFAULT Date 05 Sep 2014 13 32 Total cell concentration 8 16x10e5 cells mL Live cell concentration 6 63x10e5 cells mL Dead cell concentration 1 53x10e5 cells mL Viability 81 2 Avg size 13 8 um Total cell number 176 cells Live cell number 143 cells Dead cell number 33 cells Dilution factor 2 The Dilution Calculator initially shows the measured concentration of total cells from cell counting Users can select the Current Concentration of Total Live or Dead cells by pressing the Black Sign under the value as below Put the appropriate numbers into the blanks of the Desired Concentration and Final Volume that you want to obtain Then click Calculate in the lower right corner of screen Current Concentration 8 2 x10e 5 mL Total Desired Concentration Final Volume Calculate 53 lyo biosystems www logosbio com Chapter
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