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Summary of Contents for ThermoFisher Scientific invitrogen Countess 3
- Page 1 Countess 3 Automated Cell Counter ™ USER GUIDE Catalog Number AMQAX2000 Publication Number MAN0019566 Revision E.0 For Research Use Only. Not for use in diagnostic procedures.
- Page 2 Life Technologies Corporation | 22025 20th Ave SE Ste. 100 | Bothell, Washington 98021 USA For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition. Revision history: MAN0019566 E.0 (English) Revision Date Description 18 September Update latest draft of product manuals with latest screens and user workflow for SW#3. 2023 16 September Adding network-drive saving and Operational Qualification (OQ) functionalities.
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Page 3: Table Of Contents
Contents ■ CHAPTER 1 Product information ..........6 Contents . - Page 4 Contents ■ CHAPTER 5 Cell count and cell viability assays ....... . 22 Count cells in brightfield .
- Page 5 Contents ■ CHAPTER 9 Maintenance ............43 Instrument care .
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Page 6: Contents
Product information IMPORTANT! Before using this product, read and understand the information in the “Safety” appendix in this document. Contents The Countess 3 Automated Cell Counter is shipped with the components listed below. ™ Component Quantity Countess 3 Automated Cell Counter 1 each ™... -
Page 7: Upon Receiving The Instrument
Chapter 1 Product information Upon receiving the instrument Upon receiving the instrument Examine 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 “Install the instrument”... -
Page 8: Chapter 2 Installation
Installation Operating environment • Place the instrument on a level surface away from vibrations emanating from other pieces of equipment. • Allow at least 5 cm (2 in) free space at the back of the instrument to allow for proper ventilation and prevent overheating of electronic components. -
Page 9: Sign In To The Instrument With A New Thermo Fisher ™ Connect Platform Account
Chapter 2 Installation Sign in to the instrument with a new Thermo Fisher Connect Platform account ™ 1. Turn on the instrument by flipping the power switch on the back of the instrument to the ON position. See “Exterior instrument parts” page 7. The instrument will walk you through an initial Out-of-Box Experience to set up the instrument for first use. - Page 10 Chapter 2 Installation Sign in to the instrument with a new Thermo Fisher Connect Platform account ™ 3. Choose a method to connect. • Press Mobile device. Scan the QR code on your instrument using the Instrument Connect app on your mobile device. Follow the steps shown on the page. •...
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Page 11: Sign In To The Instrument With An Existing Thermo Fisher ™ Connect Platform Account
Chapter 2 Installation Sign in to the instrument with an existing Thermo Fisher Connect Platform account ™ Sign in to the instrument with an existing Thermo Fisher ™ Connect Platform account 1. Press (Sign in) from the Home screen. 2. Select your Username from the drop-down box. Enter your PIN. 3. -
Page 12: Chapter 3 Protocols
Protocols Protocols screen Protocols allows you to create and save customized protocols. Each custom protocol defines the count parameters (size, brightness, and circularity) and enables storing channels to be captured, calculator default values, and save settings for a consistent and streamlined workflow. •... -
Page 13: Protocol Functions
Chapter 3 Protocols Protocol functions Protocol functions The Actions screen allows you to edit, import, export, and delete protocols. Edit can be used to modify the currently selected protocol, while Import, Export, and Delete can be used on multiple protocols. Count parameters Count parameters are adjusted in the Edit Protocol screen using the Gating sliders. -
Page 14: Load A Protocol
Chapter 3 Protocols Load a protocol Load a protocol 1. Press (Protocols). 2. Press the desired protocol to select, then press Load. 3. To return to the previous screen without loading the new protocol, press Cancel or (Back). The instrument will keep the saved protocol, but will return to the previous screen without loading Note: The protocol name will be displayed in the upper left corner of the count screen. -
Page 15: Edit A Protocol
Chapter 3 Protocols Edit a protocol 5. Press Save to save the new protocol. 6. Press Cancel or (Back) to return to the Protocols screen without saving. Edit a protocol 1. Press (Protocols). 2. Select an existing protocol, then press Actions. Note: The default protocol contains default count settings and cannot be edited. 3. -
Page 16: Import A Protocol
Chapter 3 Protocols Import a protocol Import a protocol 1. Press (Protocols). 2. Press Actions. 3. Press Import. 4. Press the import location Cloud or USB. a. For Cloud, press Sign in to log in to your Thermo Fisher Connect account. If the Cloud is not active and disconnected, press Network settings to connect to a network, then press (Back) to try and sign in again. - Page 17 Chapter 3 Protocols Export a protocol 4. Select one or more protocols from the list. 5. Press Next. 6. Press the export location Cloud or USB. a. For Cloud, press Sign in to log in to your Thermo Fisher Connect account. If the Cloud is not active and disconnected, press Network settings to connect to a network, then press (Back) to try and sign in again.
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Page 18: Chapter 4 Slide Preparation And Operation
Slide preparation and operation The Countess 3 and 3 FL cell counters accept disposable Countess Cell Counting Chamber Slides ™ ™ and glass Countess Reusable Slides on interchangeable, slide-specific carriers. ™ Recommendations To obtain the best results, follow these recommendations: •... - Page 19 Chapter 4 Slide preparation and operation Load Countess Cell Counting Chamber Slide ™ 3. Let the sample mixture settle in the chamber for 30 seconds, and then insert the slide into the slide adapter. You will hear a soft click once the slide is pushed in correctly. Note: The instrument reads one side of the slide at a time, so insert the sides appropriately.
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Page 20: Load The Countess ™ Reusable Slide
Chapter 4 Slide preparation and operation Load the Countess Reusable Slide ™ Load the Countess Reusable Slide ™ 1. Before loading your sample into the Countess Reusable Slide, place a cover slip on the counting ™ chamber, making sure the cover slip is clean and free of grease. 2. - Page 21 Chapter 4 Slide preparation and operation Load the Countess Reusable Slide ™ 7. (Optional): To count the second sample present on the reusable slide, simply remove the slide from the holder, rotate, and reinsert the slide into the holder so that the second sample is aligned with the sample viewing hole.
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Page 22: Chapter 5 Cell Count And Cell Viability Assays
Cell count and cell viability assays Count cells in brightfield Rapid capture To bypass adjusting individual parameters before the brightfield count, you can select Rapid capture at the top of the Home screen. This will automatically set the parameters and perform a count as soon as the slide is inserted into the instrument. - Page 23 Chapter 5 Cell count and cell viability assays Count cells in brightfield 5. (Optional): Set exposure and/or focus. The light source slider controls the LED intensity, camera gain, and exposure time, and allows you to adjust the image brightness. The auto-focus function searches the nominal focus, or Z-point (depth) to provide fine focus to the sample.
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Page 24: Next Steps
Chapter 5 Cell count and cell viability assays Count cells in brightfield Autosaving will occur when the count is completed if you have Autosave enabled from the Save Options screen. Next steps • To identify the objects (i.e., cells) counted as Live, press . -
Page 25: View Results
Chapter 5 Cell count and cell viability assays View results View results Results screen for brightfield 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 results of the cell count and cell viability calculations (total concentration, percentage and concentration of Live and Dead cells, and percentage of Aggregates). - Page 26 Chapter 5 Cell count and cell viability assays Identify objects counted 3. To view Aggregates of cells, press on the right panel. This marks all the Aggregate cells with a yellow circle. Note: You may select one, two, or all three options. The detailed image below shows Live (green), Dead (red) cells, and Aggregates (yellow) are selected.
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Page 27: Histogram Count Results
Chapter 5 Cell count and cell viability assays Histogram count results Histogram count results View histogram For cell count and cell viability assays performed in the brightfield channel, you can view the distribution of cells (live and/or dead) based on size in a graphical format. Note: You can view the histogram on the Results and Gating screens. -
Page 28: View Scatter Plot
Chapter 5 Cell count and cell viability assays Gate count results View scatter plot For cell count and cell viability assays performed in the brightfield channel, you can view the distribution of cells (live and/or dead) based on size in a scatter plot. Note: You can view the scatter plot on the Results and Gating screens. -
Page 29: Gate Count Results
Chapter 5 Cell count and cell viability assays Save as new protocol Gate count results 1. On the Results screen, press Gating in the bottom right corner to open the Gating screen. 2. (Optional) Press (Histogram) to view the distribution of cells (live and/or dead) based on size as you gate the count results. -
Page 30: Chapter 6 Calculators
Calculators Pre-Dilution Calculator The Pre-Dilution Calculator is used to account for diluting a very concentrated sample before loading it into a sample slide for counting. For a brightfield count, this calculator also has an option to account for trypan blue use. 1. -
Page 31: Cell Splitting Calculator
Chapter 6 Calculators Cell Splitting Calculator 3. Using the keypad, enter the appropriate volumes and concentrations needed for your experiment. Press Enter after filling in both text boxes. 4. Press Calculate. The result will appear at the top of the Results screen. Cell Splitting Calculator The Cell Splitting Calculator is used to determine the amounts of sample and buffer needed to reach a desired concentration. - Page 32 Chapter 6 Calculators Cell Splitting Calculator 2. Click in one of the text fields. 3. Using the keypad, enter the appropriate volumes and concentrations needed for your experiment. Press Enter after filling in the text boxes. 4. The calculation will automatically update below the text fields. When completed, press Done. Countess 3 Automated Cell Counter User Guide ™...
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Page 33: Chapter 7 Save Results
Save results Save screen The Countess 3 and 3 FL cell counters allow you to save your data and images using a USB flash ™ drive, a link to a Thermo Fisher Connect account, or a link to a network drive. To save your experiment, choose from the following options, in any combination: •... - Page 34 Chapter 7 Save results Save procedure 2. On the Results screen, press Save to view the Save screen. 3. To assign a name to your count, press the Sample name text field and enter the name. To enter symbols, press the @%& key. To return to the keypad, tap the ABC key. 4.
- Page 35 Chapter 7 Save results Save procedure 7. To choose a different file format, press the desired file type. Available options are JPEG, PNG, and TIFF. For CSV files, you can choose from the selections of Current Count, Session, Today, Yesterday, Previous 7 days, Previous 30 days, or All. After you make your selection, the instrument returns to the Save screen.
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Page 36: Report
Chapter 7 Save results Report Report Report file The Report function allows you to save a printer-friendly report of the results, graphs, and images in PDF format. Report from brightfield count • The top section of the report contains a table with the results as displayed on the Results screen, showing the concentration of the sample and the percentage and number for the total, Live, and Dead channels, along with the percentage of Aggregates. -
Page 37: Chapter 8 Settings Overview
Settings overview Settings screen To access the Settings screen, press Settings on the Home page. Settings allows you to perform various functions: • Perform software updates (see “Update the Countess 3/3 FL Cell Counter software” page 38) ™ • Set the date and time (see “Set the date and time” page 39) Connect to a wireless network Network connecting options... -
Page 38: Software Update
Chapter 8 Settings overview Software update Software update Guidelines for software update • Software updates can be done via your Connect account. When connected to the internet, the system will automatically check for and download software updates to the instrument. Note: Using the Cloud will require a Wi-Fi dongle be installed in the instrument. •... -
Page 39: Set The Date And Time
Chapter 8 Settings overview Set the date and time 6. Select the update location. Press Next. If the Cloud option is disconnected, press Network settings to proceed to reconnect to your network. Note: Using the Cloud will require a Wi-Fi dongle be installed in the instrument. 7. - Page 40 Chapter 8 Settings overview Set the date and time 5. Press inside the Date text box to open the Edit Date keypad. 6. Using the keypad, enter the date into the Month, Day, and Year text boxes. Press Enter. 7. Press the Time text box to open the Edit Time keypad. Countess 3 Automated Cell Counter User Guide ™...
- Page 41 Chapter 8 Settings overview Set the date and time 8. Using the keypad, enter the time into the Hours and Minutes text boxes. 9. Press the drop-down menu under Time to select your time zone from the list. If needed, press Search to use a keyboard to type in your search term (e.g., pst, cst, etc.). Countess 3 Automated Cell Counter User Guide ™...
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Page 42: Change Your Cloud Region
Chapter 8 Settings overview Change your cloud region 10. Select the time format you wish to use (12 Hours with AM or PM or 24 Hours. 11. Press Apply to set the Date and Time and return to the Instrument Settings screen. Press Cancel or (Back) to return to the Instrument Settings screen without saving your changes. -
Page 43: Chapter 9 Maintenance
Maintenance Instrument care General guidelines for care • Use the appropriate cleaning solutions for each component, as indicated in the cleaning procedures in “Clean the cell counter” page 44. • If liquid spills on the instrument, turn off the power immediately and wipe dry. Power supply Always use the correct power supply. -
Page 44: Clean The Cell Counter
Chapter 9 Maintenance Clean the cell counter Clean the cell counter Introduction Clean the Countess Automated Cell Counter periodically to prevent buildup of dust and dirt that might ™ reduce its performance and cause contamination. CAUTION! To avoid electrical shock, always turn OFF the Countess Automated Cell Counter and ™... -
Page 45: Decontaminate The Instrument
Chapter 9 Maintenance Clean the cell counter Decontaminate the instrument • Wipe the instrument case of the Countess 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. •... -
Page 46: Chapter 10 Installation/Operational Qualification
Installation/Operational Qualification The installation and/or operational qualification ensures that the instrument is installed and performing within the manufacturer specifications. The qualification requires a license, thumb drive, and Countess 3 Standard Slide (Cat. No. A51876, shipped with IQ/OQ purchase). ™ Generate and activate a license 1. -
Page 47: Execute The Installation Qualification (Iq)
Chapter 10 Installation/Operational Qualification Execute the Installation Qualification (IQ) 12. Press Activate. The license is now activated and will remain valid for 2 months on the instrument. Press Next to advance to the OQ testing. Follow the OQ instructions under Execute the Operational Qualification (OQ) on page 52. - Page 48 Chapter 10 Installation/Operational Qualification Execute the Installation Qualification (IQ) 2. Enter the Protocol Execution Date, Instrument Serial Number, Lab Name, Lab Address, Contact Name, Email, and Telephone Number. 3. Select the reason (from the drop-down menu) for completing the IQ/OQ service . Countess 3 Automated Cell Counter User Guide ™...
- Page 49 Chapter 10 Installation/Operational Qualification Execute the Installation Qualification (IQ) 4. Ensure that you have access to the appropriate user guide and verify that you have read the site requirements. 5. Verify that the Service identification and Site requirement verification sections have been completed.
- Page 50 Chapter 10 Installation/Operational Qualification Execute the Installation Qualification (IQ) 6. Confirm that the automatically-filled sales order number (Document Number) is correct. 7. Verify that all of the system components are unpacked and available. Confirm that packing components are as described in the user guide. Countess 3 Automated Cell Counter User Guide ™...
- Page 51 Chapter 10 Installation/Operational Qualification Execute the Installation Qualification (IQ) 8. Verify that all required IQ tests are complete. Enter Initials and Date, then press Save. 9. Press Download to download the license to execute OQ on the instrument. Save the license to a USB drive.
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Page 52: Execute The Operational Qualification (Oq)
Chapter 10 Installation/Operational Qualification Execute the Operational Qualification (OQ) 10. Activate the OQ workflow. 11. Follow the OQ instructions in the next section, Execute the Operational Qualification (OQ), starting with step 5. Execute the Operational Qualification (OQ) Each entry throughout the OQ process is recorded in a final PDF report saved at the end of the OQ protocol. - Page 53 Chapter 10 Installation/Operational Qualification Execute the Operational Qualification (OQ) 8. Perform a series of 6 counts of the standard slide (3 counts on each side). Insert side A of the standard slide and press the appropriate count number (Perform Count 1, Perform Count 2, or Perform Count 3).
- Page 54 Chapter 10 Installation/Operational Qualification Execute the Operational Qualification (OQ) 9. Total concentration results are displayed (not editable). Pass or Fail is automatically selected based on the recorded count concentrations shown above the counts. Press Next. 10. The mean, standard deviation, and coefficient of variation (Cv) are displayed (not editable). Pass or Fail is automatically selected based on the number shown under Cv.
- Page 55 Chapter 10 Installation/Operational Qualification Execute the Operational Qualification (OQ) 14. Select the appropriate light cube from the list of available cubes. Press Next. 15. Perform a series of 6 counts of the standard slide (3 counts on each side). Insert side A of the standard slide and press the appropriate count number (Perform Count 1, Perform Count 2, or Perform Count 3).
- Page 56 Chapter 10 Installation/Operational Qualification Execute the Operational Qualification (OQ) 18. Confirm the tests are successfully completed by pressing Yes and entering your User Initials and the Date. Press Save. 19. Enter the name for the Qualification report and select the save location for the report and attachments.
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Page 57: Appendix A Troubleshooting
Troubleshooting Observation Possible cause Recommended action Abnormally high percentage of Focus was not correct for the Ensure that the cells are focused correctly so dead cells or live cells counted sample. that live cells have bright centers and dead as dead cells are dark throughout. - Page 58 Appendix A Troubleshooting Execute the Operational Qualification (OQ) Observation Possible cause Recommended action Some cells appear in the image Brightfield or fluorescence BF-based counts rely on the BF image to be but are not included in the (only in the Countess in focus. Adjust the focus and recount with ™...
- Page 59 Appendix A Troubleshooting Execute the Operational Qualification (OQ) Observation Possible cause Recommended action Variable counts for the same Pipetting was done incorrectly. If you are pipetting different samples from the sample of cells same cell sample, the variability could be due to pipetting or mixing. Use recently calibrated pipettors and make sure that the cells are well suspended by pipetting up and down several times before...
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Page 60: Appendix B Technical Specifications
Technical specifications Physical characteristics Instrument type Benchtop cell counter and suspension cell-based assay platform Instrument dimensions 9.6 (W) × 6.7 (D) × 9.4 inches (H) Weight 8 lbs Operating power 100–240 VAC, 1.0 A MAX Frequency 50/60 Hz Electrical input 12 VDC, 3 A Installation site Indoor use only, Class A Environments (i.e., non-residential or light industrial);... -
Page 61: Analysis Slide
Appendix B Technical specifications Analysis slide Analysis slide Material Poly(methyl methacrylate) (PMMA) Dimensions 25 mm (W) × 75 mm (D) × 1.7 mm (H) Chamber volume 10 µL Networking requirements Configure the system behind a firewall. If outbound traffic is limited, the following firewall exceptions are required to support system features: Firewall exception requirements Port... -
Page 62: Appendix C Countess ™ 3 Automated Cell Counter And Accessories
Countess 3 Automated Cell ™ Counter and accessories ™ The following Countess 3 instruments and instrument accessories are available from Thermo Fisher Scientific. For more information, visit www.thermofisher.com or contact Technical Support. Product Quantity Cat. No. Countess 3 Automated Cell Counter 1 each AMQAF1000 ™... -
Page 63: Appendix D Csv File Format Definition
CSV file format definition Overview A comma-separated values (CSV) file stores tabular data (numbers and text) in plain-text form. Plain text means that the file is a sequence of characters, with no data that has to be interpreted as binary numbers. - Page 64 Appendix D CSV file format definition Overview (continued) Category Column Name Description Focus motor value Motor position in relation to focal position General number BF light intensity Brightfield light intensity value from 0-100% BF LED intensity LED intensity in brightfield mode Live size min Minimum size of "live"...
- Page 65 Appendix D CSV file format definition Overview (continued) Category Column Name Description Template name Template used for count General Software revision Current software version used for count Countess 3 Automated Cell Counter User Guide ™...
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Page 66: Appendix E Safety
Safety WARNING! GENERAL SAFETY. Using this product in a manner not specified in the user documentation may result in personal injury or damage to the instrument or device. Ensure that anyone using this product has received instructions in general safety practices for laboratories and the safety information provided in this document. -
Page 67: Chemical Safety
Appendix E Safety Chemical safety Chemical safety WARNING! GENERAL CHEMICAL HANDLING. To minimize hazards, ensure laboratory personnel read and practice the general safety guidelines for chemical usage, storage, and waste provided below. Consult the relevant SDS for specific precautions and instructions: · Read and understand the Safety Data Sheets (SDSs) provided by the chemical manufacturer before you store, handle, or work with any chemicals or hazardous materials. -
Page 68: Biological Hazard Safety
Appendix E Safety Biological hazard safety Biological hazard safety WARNING! Potential Biohazard. Depending on the samples used on this instrument, the surface may be considered a biohazard. Use appropriate decontamination methods when working with biohazards. WARNING! BIOHAZARD. Biological samples such as tissues, body fluids, infectious agents, and blood of humans and other animals have the potential to transmit infectious diseases. -
Page 69: Appendix F Documentation And Support
Documentation and support Customer and technical support Visit thermofisher.com/support for the latest service and support information. • Worldwide contact telephone numbers • Product support information – Product FAQs – Software, patches, and updates – Training for many applications and instruments •... - Page 70 Countess 3 Automated Cell Counter_UG_MAN0019566-v4-GUID-E155E429- EBAF-4CA8-9CE4-7F018EEE3A3E-2022/08/15 19:02:37 en 18:16:42.969+01:00 thermofisher.com/support | thermofisher.com/askaquestion thermofisher.com 18 September 2023...
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