Cellometer® Auto 2000
Contents
1. Adjust Send Results to Printer Sample 9 Select Details to review cell images and counted cell images View Details of Counting Results 10 Select Next Sample or Assay and Settings when done Begin Next Sample 2011 Nexcelom Bioscience LLC User Interface Assay Selection Screen B Ia GE Tei ERlGreen Red cell concentration between 0 25 25 1105 cells mi Current Assay Indicates which Assay is currently selected to analyze a cell sample Edit Assay Edit the Assay settings Import Assay Import additional assays Sample ID Enter a Sample ID Dilution Factor Enter a Dilution Factor Assays Available List of Assays available for selection Preview Preview the cell sample image Load Load and analyze previously saved images Settings Edit instrument and UI settings Help Access help options Left Right Arrows Page left right to access all assays available O ON GG P OQ N eh _ oo 8 2011 Nexcelom Bioscience LLC Preview Screen Sample ID new sample Dilution Factor 2 00 AUR 7 Current View Coarse or Fine Focus Pan naar IT LE Rm o Darker Feelt ef Count Curreri Image Return lo Asia select image lo view Current Assay Indicates which Assay is currently selected to analyze a cell sample Assays amp Settings Return to Assay Selection Screen Sample ID Enter a Sample ID Dilution Factor Enter a Dilution Factor Focus Fine and coarse f2. After taking both green and red fluorescent images all fluorescent cells in each channel are counted and the concentration of live green fluorescent and dead red fluorescent cells as well as viability are determined Immune Cells Low RBC AO PI CS2 0106 or equivalent Nucleated immune cells after isolation in samples with some red blood cells PBMC Cafter ficoll separation splenocyte without lysing RBC Acridine orange is a nuclear stain that emits in the green range and is used to stain live cells Propidium iodide is a fluorescent stain that only penetrates dead cells and emits in the red range After taking both green and red fluorescent images all fluorescent cells in each channel are counted and the concentration of live green fluorescent and dead red fluorescent cells as well as viability are determined Low Concentration of Cells AO CS1 0108 Isolated primary cells or cell lines with cell concentration between 0 25 2 5 x 10 cells ml Cell Samples are prepared with acridine orange a nuclear stain that emits in the green range After taking green fluorescent images all fluorescent cells are counted and their concentration is determined Brightfield images of the sample can be taken but are not used for counting Stem Cells Primary Cells Cell Lines AO PI CS2 0106 or equivalent Stem cells primary cells cell lines cell samples from dissoc
3. contact Nexcelom 1 978 327 5340 or support nexcelom com or your local dealer After unpacking the instrument plug the Cellometer Power Cable into the back of the instrument The Auto 2000 is pre configured with the Cellometer Auto 2000 software No additional setup or configuration is required 2011 Nexcelom Bioscience LLC 13 Tutorials Overview The following tutorials are intended as a guide to performing various cell counting assays using the Auto 2000 General sample preparation hints are included for each tutorial as well as instrument and software operation instructions Each of the assays can also be performed using the sample images included in the software as a demonstration of the Auto 2000 Sample images for each cell counting assay can be found at C Program Files Nexcelom Assay_lmages Staining Solutions Trypan Blue Stock solution 0 2 in PBS Use 1 1 with cell sample Dilution factor 2 AO CS1 0108 Stock solution 10ug mlL in PBS Use 1 1 with cell sample Dilution factor 2 PI CS1 0109 Stock solution 100ug mL in PBS Use 1 1 with cell sample Dilution factor 2 AO PI CS2 0160 Stock solution 5ug mL AO 100ug mL Pl in PBS Use 1 1 with cell sample Dilution factor 2 Installed Assays and Descriptions Cell Line Total Cell Concentration No Stain Cell line or primary cells without debris with viability greater than 98 After taking brightfield images of a cell sample all cells are counted to de
4. 31 6107020 Fax 49 0 9131 6107099 e mail info peglab de www peqlab de Nexcelom Bioscience LLC 360 Merrimack Street Building 9 Lawrence MA 01843 USA Phone 1 978 327 5340 FOX 1 978 327 534 Email support nexcelom com www nexcelom com All content copyright 2011 Nexcelom Bioscience LLC Document 8001334 Rev A
5. Time Stamp Sample ID Automatically append the Sample ID with the date and time the count was performed 5 Include Instrument ID in File Automatically append the Instrument ID to the sample ID after the count is performed Auto Save 6 File File Dialog Window which allows the user to specify where the data txt file will be saved when Auto Save to Data File is selected Auto Save to Data File Automatically save the data into the Data txt file after a count is performed Create New File for Each Sample Automatically create a new data txt file for each sample and save the counting results to it after a count is performed Folder Opens File Dialog Window which allows the user to specify where the image folder will be saved when Save Raw Images or Save Counted Images is selected 2011 Nexcelom Bioscience LLC 10 Save Raw Images Automatically saves the raw images to the folder specified using the Folder command 11 Save Counted Images Automatically saves the counted images to the folder specified using the Folder command Auto Print 12 Auto Print Results Send to the default printer upon completion of count 13 Done Save Settings Save new settings and return to Assay and Settings main screen 2011 Nexcelom Bioscience LLC 19 Technical Information Specifications Environmental Requirements Typical biology lab environment Touch screen 4 Pin resistive with controller 10 4 inch format Memory RAM Memory 2 GB DDR2 Hard dri
6. een Red f EN 1 O a Sample ID new sample EERI a Dilution Factor 2 00 NI Assay Immune Cells with Low RBC Sample ID new sample Dilution Factor 2 00 Concentration Mean Diameter Total 357 cells 4 93x10 6 cells ml 6 3 microns Live 249 cells 3 44x10 6 cells ml 6 4 microns Dead 108 cells 1 49x10 6 cells ml 5 9 microns Viability 69 8 sample 7 Print aa CIE h E su CO View Details of Calculate and Adjust Send Results to Printer Begin Next Sample Counting Results Sample Current Assay Indicates which Assay is currently selected to analyze a cell sample Assays 8 Settings Return to Assay Selection Screen Sample ID Enter a Sample ID Dilution Factor Enter a Dilution Factor Results Displays cell counting results including total cell count live dead cell count and viability Details Go to the Details Screen Sample Adjustment Calculator Launch the sample adjustment calculator Useful for sample adjustment to get desired concentration or total cell number 8 Print Send the counting results to a network printer 9 Done Start a preview of a new cell sample using the same Assay settings zl O 0 P WD eh 2011 Nexcelom Bioscience LLC 11 Details Screen Sample ID new sample Vien Data Filo View Siro Histogram Current WHEE Ce Save om of Data Go Back lo Remuk Selec image lo wiew Current Assay Indicates which Assay is currently selected to analyze a cell sample Assays 8 S
7. ettings Return to Assay Selection Screen Sample ID Enter a Sample ID Dilution Factor Enter a Dilution Factor View Data File Opens data file to view or print View Size Histogram Opens separate window to display cell size histogram Save Copy of Data Saves the current counting results to the data file View Combined Image Cell image showing F1 and F2 fluorescent objects in a single merged image available only when an assay uses both Fl and F2 images 9 View Counted Image Shows cell image with green and red outlines to indicate cells counted 10 Current View Red rectangle indicates what area of the cell image is currently displayed 11 Pan Moves the area currently being shown within the current cell image 12 Zoom Zooms in or out of the current cell image 13 Select Image to Review View the brightfield Green or red fluorescence counted images 14 Return Return to Results Screen 15 Current Image Select location on slide to view counted image 0 Age o RWD 12 2011 Nexcelom Bioscience LLC Getting Started Setting up Cellometer Auto 2000 All Nexcelom products undergo a rigorous quality inspection prior to shipment and all reasonable precautions are taken in preparing them for shipment to assure safe delivery The instrument should be unpacked and inspected for mechanical damage upon receipt Mechanical inspection involves checking for signs of physical damage If damage is apparent or any components are missing please immediately
8. iated tissues with debris Acridine orange is a nuclear stain that emits in the green range and is used to stain live cells Propidium iodide is a fluorescent stain that only penetrates dead cells and emits in the red range After taking both green and red fluorescent images all fluorescent cells in each channel are counted and the concentration of live green fluorescent and dead red fluorescent cells as well as viability are determined 2011 Nexcelom Bioscience LLC Operation Reference Counting Options Use Celle Limit mama BS O Stop after 1 Images 1 Count All Use all 4 positions of slide used for cell images 2 Speed Count Have counting use Cell Limit or Image Limit a Use Cells Limit Check to stop counting after of user defined cells are counted and the next frame has finished counfing b Use Images Limit Stop counting after finishing user defined images that are less than 4 images 3 Done Save Settings Saves the current settings and returns to previous screen 2011 Nexcelom Bioscience LLC 17 Saving Options Save Options 1 Set Sample ID as Cell Type Automatically input the Sample ID to match the Cell Type parameter name being used for counting Auto Increment Sample ID Automatically append the Sample ID with an incremental numerical value Example CHO sample_001 3 Log User Name Require the user to enter in an ID that will be recorded with the data 4
9. ocus adjustment of the current cell image Exposure Adjust the exposure time of the current cell image Current View Red rectangle indicates what area of the cell image is currently displayed Pan Moves the area currently being shown within the current cell image Zoom Zooms in or out of the current cell image Select Image to Preview Select brightfield green or red fluorescence images to preview Count Starts counting of the current cell image Abort Preview Cancels current cell preview and returns Assay Selection Screen Current Image Select location on slide to preview CON OO PS Ga M ei wech _ OO N O s 2011 Nexcelom Bioscience LLC Counting Screen BRIGreen Red AO Live Cells 398 PI Dead Cells 108 Current Cound Laceby Process ounting Fluorescent 1 Image E Stop Counting Current Assay Indicates which Assay is currently selected to analyze a cell sample Assays amp Settings Return to Assay Selection Screen disabled while count is in progress Sample ID Enter a Sample ID disabled while count is in progress Dilution Factor Enter a Dilution Factor disabled while count is in progress Current Count Shows cell count numbers Counting Progress Visually indicates the current count progress Image Being Counted Indicates which image is currently being counted Stop Stops cell counting COON O VG bb ON ei 10 2011 Nexcelom Bioscience LLC Results Screen BR Gr
10. peglab Cellometer Auto 2000 User Manual 2011 Nexcelom Bioscience LLC Cellometer Auto 2000 User Manual Introduction 5 What is Cellometer Auto 2000 Si Quick Operation Instructions 5 User Interface Overview 8 Getting Started 13 Setting Up Cellometer Auto 2000 13 Tutorials 15 Overview ES Staining Solutions 15 Installed Assays and Descriptions 15 Operation Reference 17 Counting Options 17 Saving Options 18 Technical Information 21 Specifications 2l Getting Support 22 Warranty Information 22 2011 Nexcelom Bioscience LLC Introduction What is Cellometer Auto 2000 Cellometer Auto 2000 is a compact automated cell counting system utilizing dual fluorescence to detect count measure cell size and calculate cell concentration from a 20uL cell sample The basic principle of the Cellometer automatic cell counter is imaging cytometry Cells are loaded into the Disposable Counting Chamber and automatically spread into a thin layer by capillary action Cellometer Auto 2000 then captures images of cells in the counting chamber analyzes the number of cells sizes and fluorescence intensity of each cell and then converts this data into concentration size and viability The Cellometer Auto 2000 system consists of 2 main components 1 Cellometer Auto 2000 instrument with analyzing software 2 Disposable Counting Chambers Disposable counting chamber accommodates 2 individual samples and can be loaded
11. termine total cell concentration Cell Line Viability Propidium lodide PI CS1 0109 Cell line or cultured primary cells without debris Propidium iodide Is routinely used to determine cell viability Pl is a fluorescent stain that only penetrates dead cells and emits in the red range live cells are unaffected by Pl After taking both brightfield and red fluorescent images of a PI stained sample all cells from brightfield channel and dead cells from red fluorescent channel are counted to determine total and dead cell concentrations and compute the percent viability Cell Line Viability Trypan Blue Trypan Blue Cell line or cultured primary cells without debris Trypan blue is routinely used to determine cell viability Trypan blue penetrates and stains dead cells and leaves live cells unstained After taking brightfield images of the stained sample live and dead cells are counted to determine total live and dead cell concentrations as well as compute percent viability 2011 Nexcelom Bioscience LLC 15 Green Red RETS BR Green BR Green Red Immune Cells High RBC AO PI CS2 0160 or equivalent Nucleated cells in samples with large amount of red blood cells No RBC lysing Acridine orange is a nuclear stain that emits in the green range and is used to stain live cells Propidium iodide is a fluorescent stain that only penetrates dead cells and emits in the red range
12. through either port Pipette 20uL of cell sample into one of the ports with any standard single channel pipette Cellometer Auto 2000 comes with a starter set of 75 slides Slides can be ordered directly from Nexcelom or your authorized Nexcelom dealer 2011 Nexcelom Bioscience LLC 5 Quick Operation Instructions 1 Select an Assay NA Coll lino total cell conc No stain AQMP CS2 0106 or equivalent Cell line or cultured primary cells witho 7 Nudeated cels in samples with Large debas win viability greater Man 98 eat amount of red blood celts No RBC lysing 2 Input Sample ID and Dilution Factor Sample ID new sample Dilution Factor 2 00 3 Prepare Sample load disposable counting chamber and insert into instrument Cellometer Auto 4 Click Preview 5 Adjust Focus Coarse Focus 2011 Nexcelom Bioscience LLC 6 Select Green and Red images to preview and adjust exposure if needed 7 Click Count to begin counting process Count Current Image 8 Review Counting Results BR Green Red Sample ID new sample G Dilution Factor 2 00 Results Assay Immune Cells with Low RBC Sample ID new sample Dilution Factor 2 00 Concentration Mean Diameter Total 357 cells 4 93x10 6 cells ml 6 3 microns Live 249 cells 3 44x10 6 cells ml 6 4 microns Dead 108 cells 1 49x10 6 cells ml 5 9 microns Viability 69 8 Calculate and
13. ve size 8 GB compact flash BIOS 2 MB flash External Connectors 1 USB 2 0 1 Ethernet 1 Power input plug External button On off 2 color button Orange Standby Blue Ready to Count Camera inch interline 1 4 MP monochrome Pixel size 4 65 UM square 2011 Nexcelom Bioscience LLC 21 Getting Support We provide free consulting for your lab on cell type assay type and sample conditions The goal is to find the most suitable cell counting analysis solution for your lab Our team of Application Specialists are trained biologists with comprehensive understandings of cell counting viability assay methods and best practices Please contact us when you need to discuss specific types of cells sample conditions and applications We are available between 8 30am and 5 00pm Eastern US time For immediate help please call us at 1 978 327 5340 or email support nexcelom com for applications and other technical information sales nexcelom com for placing a purchase order or inquiries on a purchase order info nexcelom com for general inquiries Warranty Information Nexcelom warrants that Nexcelom instrumentation products shall for a period of 12 twelve months from the date of purchase be free of any defect in material and workmanship The sole obligation of this warranty shall be to either repair or replace at our expense the product at manufacturers option The original sales receipt must be supplied for warranty repair Products
14. which have been subjected to abuse misuse vandalism accident alteration neglect unauthorized repair or improper installation will not be covered by warranty Any Product being returned is to be properly disinfected and packaged in original packing if possible Damage sustained in shipping due to improper packing will not be covered by warranty A valid Return Material Authorization Number RMA is required for all warranty repairs For RMA instructions please contact our customer service department at 978 327 5340 or email support nexcelom com License Agreement This agreement states the terms and conditions upon which Nexcelom Bioscience LLC Nexcelom offers to license to you the software together with all related documentations The Software is licensed to you for use only in conjunction with Nexcelom s family of products Limitation of Liability Hardware and Software Cellometer branded automatic cell counting instruments software and consumables are intended for research use only In no event shall Nexcelom be liable for any damages whatsoever including without limitation incidental direct indirect special or consequential damages damages for loss of business profits business interruption loss of business information arising out of the use or inability to use this Software Consumables or related Hardware 22 2011 Nexcelom Bioscience LLC peglab Carl Thiersch Str 2b 91052 Erlangen Germany Tel 49 0 91
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