Thermo Scientific Invitrogen Countess II FL Скачать руководство пользователя страница 4

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Cell count and cell viability assays ...................... 23

Count cells in brightfield . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

Capture and count . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Next steps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

View results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Results screen for brightfield . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Identify objects counted . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Advanced

™

screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Identify live and dead cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Graph count results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

View graph . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Gate count results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Adjust screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Gate count results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Save as new protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

Edit and save profile as new protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

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CHAPTER 5

Fluorescence assays ..................................... 31

Count cell fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Count procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Next steps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

View results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

Results screen for cell fluorescence assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

Identify objects counted . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

Advanced

™

screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

Identify cells counted in fluorescence assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

Graph count results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

View graph for cell fluorescence assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

Gate count results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

Adjust screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Gate count results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

Save as new protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

Edit and save profile as new protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

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CHAPTER 6

Dilution calculator ........................................ 41

Calculate dilution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Dilution calculator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Calculate dilution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Contents

Countess

™

II FL automated cell counter User Guide

Содержание Invitrogen Countess II FL

Страница 1: ...t for use in diagnostic procedures Countess II FL automated cell counter USER GUIDE For fluorescence and brightfield applications Catalog Number AMQAF1000 Document Part Number MP10826 Publication Number MAN0010644 Revision E 0 ...

Страница 2: ... 2019 Updated trypan blue handling instructions Document converted to XML D 0 30 June 2017 Add info about edited profile indicator save profile from results screen dilution calculator and reports C 0 01 September 2015 Remove instructions for Countess II update UI for the new SW version rebrand B 0 12 December 2014 Correct technical specification for cell size A 0 08 September 2014 New user guide I...

Страница 3: ...components 11 CHAPTER 3 Getting started 12 Installation 12 Operating environment 12 Install the instrument 12 Turn ON the instrument 13 Load profile 14 Profiles screen 14 Automatic instrument functions 14 Count parameters 15 Load a profile 16 Add edit a profile 16 Display of profile names 18 Prepare sample 19 Recommendations 19 Load Countess chamber slide 19 Load Countess II FL reusable slide 20 S...

Страница 4: ...PTER 5 Fluorescence assays 31 Count cell fluorescence 31 Overview 31 Count procedure 31 Next steps 33 View results 34 Results screen for cell fluorescence assays 34 Identify objects counted 34 Advanced screen 34 Identify cells counted in fluorescence assays 35 Graph count results 36 View graph for cell fluorescence assays 36 Gate count results 37 Adjust screen 37 Gate count results 37 Save as new ...

Страница 5: ...intenance 56 Instrument care 56 General guidelines for care 56 Power supply 56 Clean the Countess II FL automated cell counter 56 Introduction 56 Clean the touch screen 57 Clean the instrument case 57 Decontaminate the instrument 57 Set nominal focus 57 Overview 57 Set nominal focus 58 APPENDIX A Troubleshooting 60 APPENDIX B Product specifications 63 Technical specifications 63 Physical character...

Страница 6: ...rument safety 74 Operating the instrument 74 Safety precautions 75 Cleaning or decontaminating the instrument 75 Removing covers or parts of the instrument 75 Chemical safety 76 Chemical waste safety 76 Chemical waste hazard 76 Chemical waste safety guidelines 77 Waste disposal 77 Electrical safety 77 Biological hazard safety 78 Safety and electromagnetic compatibility EMC standards 79 Documentati...

Страница 7: ...resources are available on the Countess Technical Resources page Go to www thermofisher com countess to access protocols application notes and tutorials Text and keyboard conventions Text and keyboard conventions used in this user guide are listed below For safety alert words and symbols used in this document see Safety alert words on page 8 Convention Use Bold Bold text indicates user action For ...

Страница 8: ...strument operation accurate installation or safe use of a chemical CAUTION Indicates a potentially hazardous situation that if not avoided may result in minor or moderate injury It may also be used to alert against unsafe practices WARNING Indicates a potentially hazardous situation that if not avoided could result in death or serious injury DANGER Indicates an imminently hazardous situation that ...

Страница 9: ...amine the instrument carefully for damage incurred during transit Ensure that all parts of the instrument including accessories listed above are included with the product Damage claims must be filed with the carrier the warranty does not cover in transit damage See Installation on page 12 for instructions on installing the instrument Visit www thermofisher com registercountess to register your ins...

Страница 10: ...licates of the same sample The instrument takes 10 seconds per sample for a typical cell count in the brightfield channel and is compatible with a wide variety of eukaryotic cells In addition to cell count and viability the Countess II FL Automated Cell Counter also provides information on cell size In addition to the brightfield channel the Countess II FL Automated Cell Counter can accommodate tw...

Страница 11: ...ess Cell Counting Chamber Slides and the glass Countess II FL Reusable Slides via interchangeable slide specific carriers For more information see Slide operation on page 20 4 Back panel The back panel of the Countess II FL Automated Cell Counter allows access to the optional EVOS light cubes and provides storage for the light cube tool and the reusable slide carrier The back panel is secured to t...

Страница 12: ...rn off the main power switch located on the back of the instrument Instrument exterior components on page 11 In case of an instrument malfunction turn the main power switch to the OFF position and disconnect the instrument from the wall outlet 1 Unpack the instrument and place the instrument on a flat level dry surface 2 Remove the thin plastic protector film from the touch screen display 3 Plug o...

Страница 13: ...rrent profile or to add a new profile to the instrument press the Profiles button in the upper left corner Profiles allow you to create customized count preferences i e gate counts based on cell size brightness circularity and or relative fluorescence intensity Load profile on page 14 4 To change instruments settings press the Instrument Settings button in the upper right corner Instrument setting...

Страница 14: ...loading a new profile from the Results screen applies the count preferences to the current counts results total cells viability etc and to all new counts If you change any setting that is saved as part of the protocol size brightness or circularity on the Results screen the profile name is appended with the symbol You can turn the Auto FL Threshold and Auto Lighting functions ON and OFF using the ...

Страница 15: ...ted Fluorescence intensity As you move the slider up the algorithm includes the objects that fluoresce more brightly in the count As you move the slider down only the dimmest of objects are counted Size brightness and fluorescence intensity sliders are range sliders To adjust the upper and lower boundaries without changing the data range drag the slider by its middle section i e the slider bar To ...

Страница 16: ... opens Note The Default profile contains default count settings and cannot be edited 3 Select or deselect the Auto FL Threshold checkbox to turn the Auto FL Threshold function ON or OFF Automatic instrument functions on page 14 Note This function is available only for instruments equipped with the optional EVOS light cubes 4 Select or deselect the Auto Lighting checkbox to turn the Auto Lighting f...

Страница 17: ...eters on page 15 Parameter sliders in the BF channel allow you to gate count results based on size brightness and circularity The parameter slider in the selected fluorescence channel allows you to gate count results based on relative fluorescence intensity in that channel c Repeat for the remaining channels as necessary Note You can further adjust the size brightness or circularity parameters for...

Страница 18: ...to the previous screen 7 To return to the previous screen without loading the new profile press the previous button The instrument will keep the saved profile but return to the previous screen without loading it The maximum display length for profile names on the screen is seven characters If the profile name is over seven display characters the name is shortened to the first seven characters with...

Страница 19: ...can be reduced by gentle heating at 37 C for 10 minutes 1 Prepare the sample by adding 10 µL of your cell suspension to 10 µL of 0 4 trypan blue stain Mix the sample mixture well by pipetting it up and down a few times 2 Gently pipet 10 µL of the sample into the half moon shaped sample loading area The sample is loaded into the chamber through capillary action 3 Let the sample mixture settle in th...

Страница 20: ... Cell Counting Chamber Slide or the Countess II FL Reusable Slide has a 10 µL sample capacity Do not overfill the slide chambers Slide operation The Countess II FL instrument accepts both disposable Countess Cell Counting Chamber Slides and glass Countess II FL Reusable Slides on interchangeable slide specific carriers 1 To use the plastic disposable Countess Cell Counting Chamber Slide with the C...

Страница 21: ...squeeze the tabs and pull the carrier completely out of the instrument Note You can store the slide carrier behind the access panel on the back of the instrument Instrument exterior components on page 11 1 To use the Countess II FL Reusable Slide unlatch the back panel of the Countess II FL Automated Cell Counter with the two captive turn fasteners that secure the back panel on the rear of the ins...

Страница 22: ...ument until it clicks and a spring pushes the slide out Grasp the slide and pull it out the rest of the way 6 Optional To count the second sample present on the reusable slide simply remove the slide from the carrier rotate and reinsert the slide into the carrier so that the second sample is aligned with the sample viewing hole Note You can store the slide carrier behind the access panel on the ba...

Страница 23: ...e slide into the slide port Instrument exterior components on page 11 making sure that the sample side is inserted completely into the instrument You will hear a soft click if the slide is pushed in correctly 5 When the slide is inserted the instrument automatically illuminates the sample sets the intensity of brightfield illumination and auto focuses on the cells Note To turn off the Auto Lightin...

Страница 24: ...pped with an EVOS light cube first press Adjust and then select brightfield white circle as the light source Set the exposure then press Done to return to the Capture screen 9 Press Capture Note If your instrument is equipped with an EVOS light cube make sure that only the BF brightfield checkbox is selected under Collect channels before capturing the image The instrument temporarily captures the ...

Страница 25: ...cation Chapter 6 Dilution calculator To permanently save the results press Save Chapter 7 Save results To perform a new count remove the slide and reinsert it second chamber first into the instrument or insert a new sample slide View results The Results screen for cell count and cell viability assays performed using the brightfield channel displays a composite image of the objects counted and the ...

Страница 26: ...button Live cells will be circled in green on the screen To identify the cells that are included in the count as dead press the Dead toggle button Dead cells will be circled in red on the screen Note You may select either or both options In the following example both Live and Dead buttons are pressed and live and dead cells are marked with green and red circles respectively 3 To unmark the cells i...

Страница 27: ...t parameters As you adjust the count parameters the count results and the graph will be automatically updated 4 To close the graph press the Graph button again Gate count results The Adjust screen for cell count and cell viability assays in the brightfield channel contains the controls for gating results based on size brightness and circularity You can adjust the count parameters before or after p...

Страница 28: ... Count parameters on page 15 7 When finished press Done to save the changes to count parameters and return to the Advanced screen Press Cancel to return to the Results screen without saving the changes 8 On the Advanced screen press Count to recalculate your results with the new count parameters 9 To save the changes to size brightness or circularity parameters to the current profile or to create ...

Страница 29: ...ofile with the edited parameters press the New protocol button The Select profile to edit screen opens Note By default the current profile button is selected on the Select profile screen If you are using the Default profile for the count no profile button is selected on this screen 3 Select the profile you wish to edit then press Import settings Note You can select only a previously saved or an em...

Страница 30: ...names on page 18 6 Optional If desired make additional changes to the profile and the count parameters as described in Add edit a profile on page 16 7 Click Save to save the new profile settings and return to the Results page for the last count The profile name will be displayed without the symbols Click Cancel to return to the Results page for the last count without saving the changes to the prof...

Страница 31: ...amber slide on page 19 Let the sample mixture settle for 30 seconds 3 Optional Press the Profiles button located on the upper left corner of the screen to open the Profiles screen and load the desired profile as described in Load profile on page 14 4 Insert the sample slide into the slide port Instrument exterior components on page 11 making sure that the sample side is inserted completely into th...

Страница 32: ... the desired Light source button The instrument displays the sample in the selected channel brightfield or fluorescent In the example below the sample is displayed in the GFP channel Note The light source buttons select the light channel brightfield and or fluorescence for sample illumination and are used when setting the exposure for the selected channel Steps 9 on page 32 11 on page 33 they do n...

Страница 33: ...y objects counted on page 34 To see the distribution of cells counted through each channel in a graphical format press the Graph button Graph count results on page 36 To gate the results by object size brightness circularity or relative fluorescence intensity first press More to open the Advanced screen then press Adjust to go to the Adjust screen Gate count results on page 37 Note You can save th...

Страница 34: ...ell concentration in the sample slide which must be multiplied by any dilution factor present to calculate the original cell concentration This is in contrast to the cell counts in brightfield where the counting algorithm assumes a 1 1 dilution of the sample in trypan blue and displays the original cell concentration i e before the dilution in the Results screen Identify objects counted The Advanc...

Страница 35: ...esponding boundaries button Cells counted in the selected channel will be circled on the screen with the same color as the selected channel In the example below both the GFP and TxRed boundaries buttons are pressed and the cells counted in the GFP and TxRed channels are marked with green and red circles respectively 4 To unmark the cells counted in a specific channel press the corresponding bounda...

Страница 36: ...elect BF brightfield The graph displays the size distribution of the total cell count number of cells vs cell size in µm and the average size of the cells counted in each available fluorescence channel 2 To view the distribution of cells based on relative fluorescence intensity press the Graph button and then select FL fluorescence The graph displays the distribution of cells based on fluorescence...

Страница 37: ...luorescence intensity You can adjust the count parameters before or after performing a count and save these changes to the current profile or as a separate profile Save as new protocol on page 39 1 On the Results screen press More to open the Advanced screen 2 Optional To view your sample under a specific light source brightfield and or fluorescent select the desired Channels checkbox on the Advan...

Страница 38: ...desired fluorescence channel colored circles to adjust the threshold for fluorescence intensity using the fluorescence intensity slider Note The fluorescence channels available depend on the EVOS light cubes installed in the instrument 8 When finished press Done to save the changes to count parameters and return to the Advanced screen Press Cancel to return to the Results screen without saving the...

Страница 39: ...s press the New protocol button The Select profile to edit screen opens Note By default the current profile button is selected on the Select profile screen If you are using the Default profile for the count no profile button is selected on this screen because the Default profile cannot be edited 3 Select the profile you wish to edit then press Import settings Note You can select only a previously ...

Страница 40: ...page 18 6 Optional If desired make additional changes to the profile and the count parameters as described in Define count parameters for the fluorescent channels on page 17 7 Click Save to save the new profile settings and return to the Results page for the last count The profile name will be displayed without the symbols Click Cancel to return to the Results page for the last count without savin...

Страница 41: ...ress the cell type button located to the right of the current concentration then select the count result you wish to use for the dilution calculation from the dropdown The current concentration changes to reflect the results for the cell type selected Available Cell type options are For cell count and viability assays in brightfield Live cells or Total cells By default Live cells is selected 6 Dil...

Страница 42: ...imal separator integer part and one to the right fractional part If you do not enter the fractional part the software enters a 0 by default Press Enter or touch anywhere outside the number pad to close it b Press the exponent button then select the exponent value for the desired concentration By default the exponent is n 1 where n the exponent value of the current count The maximum selectable expo...

Страница 43: ...ume and closed the last popup window the bottom line of the dilution calculator displays the volumes of cell solution and buffer needed If you enter a combination of values that is not valid e g desired concentration is greater than the current concentration the results line remains on or return to the blank state and screen displays a warning message If you make any changes to any of the input ar...

Страница 44: ...program For more information on the CSV file format see Appendix D CSV file format Report Saves a printer friendly report of the results graph s and image in the selected format PDF PNG or JPEG For more information see Report on page 47 Note If you wish to save your results with the Graph showing the distribution of cells based on cell size or fluorescence intensity make sure that the desired grap...

Страница 45: ...er the file name using the alpha numeric keypad To enter symbols press the symbol key To return to the alpha numeric keypad press ABC 5 Press Enter to save the name and return to the Save screen To return to the Save screen without saving the name press the close button Chapter 7 Save results Save count results 7 Countess II FL automated cell counter User Guide 45 ...

Страница 46: ...nd Report is saved as PDF To choose a different file format press the file type button The Choose file type screen opens Note Data can only be saved as a CSV file 8 Press to select the desired file type Available options are JPEG BMP PNG and TIFF After you make your selection the instrument returns to the Save screen To return to the Save screen without changing the file format press the close but...

Страница 47: ...ocation Report The Report function allows you to save a printer friendly report of the results graphs and images in the selected format PDF PNG or JPEG You can create reports using the Report dropdown as described in Save count results on page 44 Report file Chapter 7 Save results Report 7 Countess II FL automated cell counter User Guide 47 ...

Страница 48: ...l Live and Dead channels Below the results table the report contains the number of cells vs cell size graph Under the graph the report contains the brightfield count image with the live and dead cells identified by the green and red circles respectively At the bottom the report displays the profile information used to gate these images Chapter 7 Save results Report 7 48 Countess II FL automated ce...

Страница 49: ... graphs the report contains the count images with the brightfield image on the left and the fluorescence images on the right In the brightfield image the cells counted in the brightfield channel are identified by the white total count circles In the fluorescence image overlaid if there are two channels the cells counted are identified by the circles with the same color as the fluorescence channel ...

Страница 50: ...e file must be FAT32 formatted verify this before proceeding If necessary reformat the USB drive to FAT32 following the recommended procedure for your operating system Note Reformatting the USB drive will result in the loss of all files Back up the files in the USB drive prior to reformatting The software update file must be saved on the top level of the USB drive not within a folder or a subfolde...

Страница 51: ... USB drive into one of the USB ports of the instrument Instrument exterior components on page 11 4 Press the Instrument Settings button on the Home page Turn ON the instrument on page 13 to open the Instrument Settings screen Chapter 8 Instrument settings 5 Select Software Update from the Instrument Settings menu The instrument scans the USB drive for the latest software version 6 When prompted se...

Страница 52: ...ct the Date format you wish to use 4 Press any Date text box MM DD or YYYY to open the Edit Date keypad 5 Using the keypad enter the date into Month Day and Year text boxes pressing Enter after each entry 6 After you are finished entering the date press the close button to return to the Date Time screen Set the date and time Chapter 8 Instrument settings Date Time 8 52 Countess II FL automated cel...

Страница 53: ...ter after each entry 10 After you are finished entering the time press the close button to return to the Date Time screen 11 If you have selected the 12 Hour format select AM or PM 12 Press Save to set the Time and Date and return to the Instrument Settings screen Press Cancel to return to the Instrument Settings screen without saving your changes Chapter 8 Instrument settings Date Time 8 Countess...

Страница 54: ...Light Cube The instrument positions the light cube tray to enable light cube installation 3 When prompted power off the Countess II FL Automated Cell Counter using the power switch on the back of the instrument Instrument exterior components on page 11 4 Unplug the power cord from the Countess II FL Automated Cell Counter 5 Unlatch the back panel with the two captive turn fasteners indicated by re...

Страница 55: ...ws in Figure B and gently pull the light cube out of the device Note Always store the cube removal tool in the back panel for easy access 10 Install the back panel and secure it in its place with both turn fasteners 11 Plug the power cord back into the Countess II FL Automated Cell Counter 12 Turn off the Countess II FL Automated Cell Counter by flipping the power switch on the back of the instrum...

Страница 56: ...range for service IMPORTANT If you have any doubt about the compatibility of decontamination or cleaning agents with parts of the equipment or with material contained in it contact Technical Support or your local distributor for information Clean the Countess II FL automated cell counter Clean the Countess II FL Automated Cell Counter periodically to prevent buildup of dust and dirt that might red...

Страница 57: ... the instrument is plugged in Wipe the instrument case of the Countess II FL Automated Cell Counter using a soft lint free cloth moistened with 70 alcohol Wipe the instrument dry immediately after cleaning Avoid using a bleach solution because it may leave a residue of bleach crystals on the instrument Ensure that water or other cleaning solutions do not enter the power button the power inlet the ...

Страница 58: ...ide Load Countess chamber slide on page 19 or the Countess II FL Reusable Slide Countess II FL reusable slide on page 21 Let the sample mixture settle for 30 seconds to ensure a uniform focal plane 3 Insert the sample slide into the slide port of the instrument Instrument exterior components on page 11 making sure that the side containing the sample is inserted completely 4 When the slide is inser...

Страница 59: ... to adjust focus or lighting 7 After finding the optimal focus press Set to set the nominal focus Once the focus has been set the Set button on the focus slider becomes inactive confirming that the focus setting has been stored Chapter 9 Maintenance Set nominal focus 9 Countess II FL automated cell counter User Guide 59 ...

Страница 60: ...page 57 Setting the nominal focus will improve autofocus consistency with future slides To set the nominal focus see Set nominal focus on page 57 Some cells appear on the image but are not included in the count For cell count and cell viability assays performed in the brightfield adjust the size brightness and circularity gates for both live and dead cells to include all of the cells in the count ...

Страница 61: ...dding trypan blue Pipet the bead and trypan blue mixture up and down several times to make sure it is well mixed and load 10 µL into the slide without delay Variable counts when performing replicate counts of the same slide If you are counting replicates of the exact same slide visually inspect that all cells are counted correctly in the image There may be a slightly different field of view each t...

Страница 62: ...he debris using the size brightness and circularity sliders USB drive not recognized by the instrument The USB drive must be FAT32 formatted to be recognized by the instrument If it is not reformat the USB drive to FAT32 Software update on page 50 Try another correctly formatted USB drive Unable to update the Countess software Make sure the USB drive is formatted to FAT32 If it is not reformat the...

Страница 63: ...Operating humidity 80 non condensing Processing time 15 seconds Sample concentration range 1 104 1 107 cells mL Particle cell diameter range 4 60 µm particles 7 60 µm cells Required sample volume 10 µL Firmware Countess Automated Cell Counting Platform Software USB Drive 4 Gigabytes Optics 3 channels brightfield and 2 slots for EVOS LED light cubes Camera 5 Mega pixels 2 5 Optical Magnification Ma...

Страница 64: ...scent light cubes for multiple fluorescence research applications The following table lists some of the common fluorescent and specialty EVOS light cubes available from Thermo Fisher Scientific For a complete list go to www thermofisher com evoslightcubes or contact Technical Support For instructions on changing the LED light cubes see Change light cube on page 54 Light cube Dye DAPI DAPI Hoechst ...

Страница 65: ...untess II FL Automated Cell Counter The Countess II Automated Cell Counter uses only brightfield illumination and does not support the EVOS light cubes Appendix B Product specifications EVOS light cubes B Countess II FL automated cell counter User Guide 65 ...

Страница 66: ...de Holder 1 each AMEP4746 Accessory products The following products can be used with the Countess II FL Automated Cell Counter and are available separately from Thermo Fisher Scientific For more information visit www thermofisher com or contact Technical Support Product Quantity Cat No Countess Cell Counting Chamber Slides 50 Slides 100 counts 1 box 1 C10228 Countess Cell Counting Chamber Slides 5...

Страница 67: ...Trypan blue stain 0 4 2 1 mL T10282 1 Each box of Countess Cell Counting Chamber Slides contains 50 slides and 2 1 mL vials of trypan blue 0 4 sufficient for 100 counts Appendix C Ordering information Accessory products C Countess II FL automated cell counter User Guide 67 ...

Страница 68: ...tal cells counted Total number of cells counted in the sample G Live concentration Concentration of just the live portion of the sample H Live cells counted Total number of live cells counted I Dead concentration Concentration of just the dead portion of the sample J Dead cells counted Total number of dead cells counted K Viability Percent viability of the sample based on trypan blue staining L Av...

Страница 69: ... min Minimum size of live cells in microns AA Live Size max Maximum size of live cells in microns AB Live Brightness min Live adjustment slider value for minimum brightness AC Live Brightness max Live adjustment slider value for maximum brightness AD Live Circularity Live adjustment slider value for circularity AE Dead Size min Minimum size of dead cells in microns AF Dead Size max Maximum size of...

Страница 70: ...ity AQ Cube 1 Brightness min First top light cube adjustment slider value for minimum brightness AR Cube 1 Brightness max First top light cube adjustment slider value for maximum brightness AS Cube 2 Brightness min Second bottom light cube adjustment slider value for minimum brightness AT Cube 2 Brightness max Second bottom light cube adjustment slider value for maximum brightness Appendix D CSV f...

Страница 71: ...e aware of relevant hazards Each alert word IMPORTANT CAUTION WARNING DANGER implies a particular level of observation or action as defined below IMPORTANT Provides information that is necessary for proper instrument operation accurate installation or safe use of a chemical CAUTION Indicates a potentially hazardous situation that if not avoided may result in minor or moderate injury It may also be...

Страница 72: ...ected to earth ground before any other electrical connections are made to the instrument Indicates a terminal that can receive or supply alternating current or voltage Indicates a terminal that can receive or supply alternating or direct current or voltage Safety symbols The following table describes the safety symbols that may be displayed Each symbol may appear by itself or in combination with t...

Страница 73: ...13 2005 Symbol Description Do not dispose of this product as unsorted municipal waste Follow local municipal waste ordinances for proper disposal provisions to reduce the environmental impact of waste electrical and electronic equipment WEEE European Union customers Call your Customer Service representative for equipment pick up and recycling See www thermofisher com for a list of customer service...

Страница 74: ... pas retirer les capots dont l ouverture nécessite l utilisation d outils L instrument ne contient aucune pièce réparable par l utilisateur Toute intervention doit être effectuée par le personnel de service qualifié venant de Thermo Fisher Scientific DANGER Class 3B visible and or invisible laser radiation present when open Avoid exposure to beam DANGER Rayonnement visible ou invisible d un faisce...

Страница 75: ... sure that the power cord is properly grounded Be sure to position the equipment such that it is easy to disconnect the instrument Turn off the instrument before unplugging the power cord and or moving the instrument If the instrument is broken or dropped disconnect the power cord and contact a service person Do not disassemble the instrument Use only authorized accessories adaptor power cord and ...

Страница 76: ... contains spills or leaks from the primary container Both containers must be compatible with the waste material and meet federal state and local requirements for container storage After emptying a waste container seal it with the cap provided Characterize by analysis if needed the waste generated by the particular applications reagents and substrates used in your laboratory Ensure that the waste i...

Страница 77: ...practices and local state provincial or national environmental and health regulations If potentially hazardous waste is generated when you operate the instrument you must Characterize by analysis if necessary the waste generated by the particular applications reagents and substrates used in your laboratory Ensure the health and safety of all personnel in your laboratory Ensure that the instrument ...

Страница 78: ...ad and follow the guidelines in these publications ATTENTION BIOHAZARD Les échantillons biologiques tels que les tissus les fluides corporels et le sang des humains et d autres animaux ont la possibilité de transmettre des maladies infectieuses Suivre tous les règlements municipaux provinciaux provincial et ou nationales en vigueur Porter des lunettes de protection approprié des vêtements et des g...

Страница 79: ...m CSA CSA America ANSI ASME ASSE ASTM NSF and UL European safety and EMC standards The CE Mark symbolizes that the product conforms to all applicable European Community provisions for which this marking is required Operation of the instrument is subject to the conditions described in this manual The protection provided by the instrument may be impaired if the instrument is used in a manner not spe...

Страница 80: ... protocols Certificates of Analysis Safety Data Sheets SDSs also known as MSDSs Note For SDSs for reagents and chemicals from other manufacturers contact the manufacturer Limited product warranty Life Technologies Corporation and or its affiliate s warrant their products as set forth in the Life Technologies General Terms and Conditions of Sale at www thermofisher com us en home global terms and c...

Страница 81: ......

Страница 82: ...thermofisher com support thermofisher com askaquestion thermofisher com 7 June 2019 ...

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