Related Manuals for Thermo Scientific Invitrogen Countess 3 FL
Summary of Contents for Thermo Scientific Invitrogen Countess 3 FL
- Page 1 ™ Countess 3 FL Automated Cell Counter USER GUIDE Catalog Number AMQAF2000 Publication Number MAN0019567 Revision B.0 For Research Use Only. Not for use in diagnostic procedures.
- Page 2 Life Technologies Corporation | 22025 20th Ave SE Ste. 100 | Bothell, WA 98021 For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition. The information in this guide is subject to change without notice. DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
-
Page 3: Table Of Contents
Contents ■ CHAPTER 1 Product information ..........7 Contents . - Page 4 Contents ■ CHAPTER 5 Cell count and cell viability assays ....... . 24 Count cells in brightfield .
- Page 5 Contents ■ CHAPTER 8 Save results ............41 Save screen .
- Page 6 Contents ■ ™ APPENDIX D Countess 3 FL Automated Cell Counter and accessories . . . 63 Accessory products ............63 ■...
-
Page 7: Chapter 1 Product Information
Product information IMPORTANT! Before using this product, read and understand the information in the “Safety” appendix in this document. Contents ™ The Countess 3 FL Automated Cell Counter is shipped with the components listed below. Component Quantity ™ Countess 3 FL Automated Cell Counter 1 each Power cord with 4 adapters (for U.S./Canada/Taiwan/Japan, Europe, or UK) 1 each... -
Page 8: Product Description
Chapter 1 Product information Product description Product description ™ The Countess 3 FL Automated Cell Counter is a fully automated, 3-channel cell counter and assay ™ platform that uses EVOS light cube technology, state-of-the-art optics, and image analysis algorithms to analyze fluorescently labeled cells or trypan blue stained samples in suspension. ™... -
Page 9: Exterior Instrument Parts
Chapter 1 Product information Exterior instrument parts Exterior instrument parts ™ Touchscreen display: The 7-inch capacitive display is the main user interface of the Countess 3 FL Automated Cell Counter. It contains the buttons for all instrument functions and displays data from the cell count. Front USB port: Allows you to transfer and save the cell count data and image to an external computer for record keeping and printing purposes. -
Page 10: 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 11: Sign In To The Instrument With A New Connect Account
Chapter 2 Installation Sign in to the instrument with a new Connect 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” on page The instrument will walk you through an initial Out-of-Box Experience to set up the instrument for first use. - Page 12 Chapter 2 Installation Sign in to the instrument with a new Connect 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. •...
-
Page 13: Sign In To The Instrument With An Existing Connect Account
Chapter 2 Installation Sign in to the instrument with an existing Connect account Sign in to the instrument with an existing Connect account 1. Press Sign in from the Home screen. 2. Select your Username from the drop-down box. Enter your PIN. 3. -
Page 14: Chapter 3 Templates
Templates Templates screen Templates allows you to create and save customized templates. Each custom template defines the count parameters (size, brightness, circularity, and fluorescence intensity) for a consistent and streamlined workflow. • Templates can be accessed from the Home and Gating screens. •... -
Page 15: Template Functions
Chapter 3 Templates Template functions Template functions The Actions screen allows you to edit, import, export, and delete templates. Edit can be used to modify the currently selected template, while Import, Export, and Delete can be used on multiple templates. Count parameters Count parameters are adjusted in the Edit template screen using the Gating sliders. -
Page 16: Load A Template
Chapter 3 Templates Load a template Load a template 1. Press Templates. 2. Press the desired template to select, then press Load. 3. To return to the previous screen without loading the new template, press Cancel or Back. The instrument will keep the saved template, but will return to the previous screen without loading Create a template 1. -
Page 17: Import A Template
Chapter 3 Templates Import a template 3. Press Edit. The Edit screen for the selected template opens. 4. Press the appropriate channel (BF or other dye) and edit the desired parameters (see “Count parameters” on page 15) to create a custom template. 5. -
Page 18: Export A Template
Chapter 3 Templates Export a template b. For USB, insert your USB drive into the slot on the front of the instrument. 5. Press Next. 6. Select one or more templates from the list. 7. Press Import. 8. Once complete, press Done. Export a template 1. -
Page 19: Delete A Template
Chapter 3 Templates Delete a template b. For USB, insert your USB drive into the slot on the front of the instrument. 7. Press Export. 8. Once complete, press Done. Delete a template 1. Press Templates. 2. Press Actions. 3. Press Delete. 4. -
Page 20: 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 21: Load The Countess ™ Reusable Slide
Chapter 4 Slide preparation and operation Load the Countess ™ Reusable 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. - Page 22 Chapter 4 Slide preparation and operation Load the Countess ™ Reusable Slide 3. Gently pipet 10 µL of the sample into the sample inlet, allowing capillary action to draw the sample into the counting chamber. A properly loaded counting chamber should have a thin, even film of fluid under the cover slip.
- Page 23 Chapter 4 Slide preparation and operation Load the Countess ™ Reusable Slide 5. Remove the reusable slide holder from inside of the back panel. 6. Load the reusable glass slide into the reusable slide holder. 7. Insert the reusable slide holder containing the slide into the slide port and gently push into the instrument until it clicks into place.
-
Page 24: Chapter 5 Cell Count And Cell Viability Assays
Cell count and cell viability assays Count cells in brightfield Capture and count 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 25: Next Steps
Chapter 5 Cell count and cell viability assays Count cells in brightfield ™ ™ Note: For the Countess 3 FL Automated Cell Counter equipped with an EVOS light cube, first press Adjust, and then select BF (brightfield) as the light source. Set the exposure, then press Apply to return to the Setup screen. -
Page 26: 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). -
Page 27: Identify Cells Counted In Cell Count And Cell Viability Assays
Chapter 5 Cell count and cell viability assays Identify objects counted Identify cells counted in cell count and cell viability assays On the Results screen, Live or Dead cells can be selected. 1. To view Live cells, press on the right panel. This marks all Live cells in green. 2. -
Page 28: 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 29: Gate Count Results
Chapter 5 Cell count and cell viability assays Save as new template 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 Fluorescence Assays
Fluorescence assays Count cell fluorescence Overview ™ ™ Countess 3 FL Automated Cell Counter equipped with the optional EVOS light cubes can be used for a variety of fluorescent applications, including simultaneous counts of cells stained with two different fluorescent dyes, GFP and RFP expression, and apoptosis and cell viability assays. ™... - Page 31 Chapter 6 Fluorescence assays Count cell fluorescence 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 9-11); they do not determine which channels are used for capturing the image. 7.
- Page 32 Chapter 6 Fluorescence assays Count cell fluorescence 10. On the Setup screen, select the check boxes to the right of the channels you wish to capture. BF (brightfield) is checked by default and cannot be deselected. 11. Press Count. The instrument captures the images and displays the results (total concentration, percentage and concentration of cells counted in each fluorescence channel).
-
Page 33: Next Steps
Chapter 6 Fluorescence assays View results Next steps • To identify the brightfield objects (i.e., cells), press . For identifying fluorescence objects, press ™ the appropriate colored icon for the specific EVOS light cube (e.g., press for GFP, press TX Red). •... -
Page 34: Identify Objects Counted
Chapter 6 Fluorescence assays Identify objects counted 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. - Page 35 Chapter 6 Fluorescence assays Histogram count results 2. To view the distribution of cells based on relative fluorescence intensity, press Histogram, then select FL (fluorescence). The histogram displays the distribution of cells based on fluorescence intensity. 3. To remove the cells counted in a specific channel from the histogram, press the corresponding channel button on the histogram.
-
Page 36: Gate Count Results
Chapter 6 Fluorescence assays Gate count results Gate count results Gating screen The Gating screen for cell fluorescence assays contains the controls for gating count results based on size, brightness, circularity, and fluorescence intensity. You can adjust the count parameters before or after performing a count, and save these changes to the current template or as a separate template (“Save as new template”... -
Page 37: Save As New Template
Chapter 6 Fluorescence assays Save as new template Save as new template Edit and save a new template If you have made changes to the count parameters after performing a count, you may save your settings to a new template. 1. -
Page 38: Chapter 7 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 39: Cell Splitting Calculator
Chapter 7 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 40 Chapter 7 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 ™...
-
Page 41: Chapter 8 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 or link to a Thermo Fisher Connect account. To save your experiment, choose from the following options, in any combination: •... - Page 42 Chapter 8 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 symbol (@%&) key. To return to the keypad, press ABC. 4.
- Page 43 Chapter 8 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 Capture, Session, Yesterday, Past 3 days, Past 7 days, or All. After you make your selection, the instrument returns to the Save screen.
-
Page 44: Report
Chapter 8 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. - Page 45 Chapter 8 Save results Report Report from fluorescence 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 of cells for the total, FL1, FL2, and FL1 + FL2 channels.
-
Page 46: Chapter 9 Settings Overview
Settings overview Settings screen To access the Settings screen, press Settings on the Home page. Settings allows you to: • Perform software updates (see “Update the Countess ™ 3/3 FL Cell Counter software” on page • Set the date and time (see “Set the date and time” on page ™... -
Page 47: Software Update
Chapter 9 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 48: Set The Date And Time
Chapter 9 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 49: Change Your Cloud Region
Chapter 9 Settings overview Change your cloud region 7. Press any Time text box (Hours or Minutes) to open the Edit Time keypad. 8. Using the keypad, enter the time into the Hours and Minutes text boxes. 9. Select the time format you wish to use (12 Hours with AM or PM or 24 Hours. 10. - Page 50 Chapter 9 Settings overview Change your cloud region 3. Press Cloud region. Your current cloud region is displayed at the bottom of the screen. 4. Select the desired cloud region from the available choices. Press Change. 5. Once the change is complete, press Done. Note: Changing the cloud region will require relinking of all instrument accounts.
-
Page 51: Chapter 10 Change Evos ™ Light Cubes
™ Change EVOS light cubes ™ Install or change an EVOS light cube ™ ™ The Countess 3 FL Automated Cell Counter can accommodate up to two EVOS light cubes. Each ™ user-interchangable, auto-configured EVOS light cube contains an LED, collimating optics, and filters ™... - Page 52 Chapter 10 Change EVOS ™ light cubes ™ Install or change an EVOS light cube 7. Place the light cube into one of the empty slots in the back of the device. 8. Using the tool provided on the inside of the back panel (red arrow in Figure A), secure the light cube by tightening the two screws on the end of the cube (red arrows in Figure B).
-
Page 53: Chapter 11 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” on page • If liquid spills on the instrument, turn off the power immediately and wipe dry. Power supply Always use the correct power supply. -
Page 54: Clean The Cell Counter
Chapter 11 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 unplug the power cord before cleaning or decontaminating the instrument. -
Page 55: Decontaminate The Instrument
Chapter 11 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 56: Appendix A Troubleshooting
Troubleshooting Observation Possible cause Recommended action Uneven screen illumination Light cubes misaligned. Reset the light cube tray by selecting Change (screen is dark on one side, light cube from the Instrument Settings screen. bright on the other side) See Appendix C, “EVOS light cubes”. Autofocus does not focus Debris or other material Make sure there are no bubbles or debris visible... - Page 57 Appendix A Troubleshooting Clean the cell counter Observation Possible cause Recommended action Incorrect concentration for the Beads were not suspended in The beads can settle quickly in solution, which ™ Countess test beads solution or mixed correctly. will affect the concentration reading. Vortex the bead stock on high for a full 30 seconds to resuspend, and add 10 µL of the bead suspension to 10 µL of trypan blue without...
- Page 58 Appendix A Troubleshooting Clean the cell counter Observation Possible cause Recommended action Abnormally high percentage of Capture settings were not Gate out the debris using the size, brightness, dead cells or live cells counted optimal. and circularity sliders. as dead (continued) USB drive not recognized by the USB drive was not correctly...
-
Page 59: 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 60: 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 Countess ™ 3 FL Automated Cell Counter User Guide... -
Page 61: Appendix C Evos Light Cubes
EVOS light cubes LED illumination ™ The Countess 3 FL Automated Cell Counter utilizes an adjustable intensity LED light source provided ™ by the proprietary, user-interchangeable LED light cube (see “EVOS light cubes” on page 61). Because the LED light source is as close as possible to the objective, the number of optical elements in the channel is minimized. - Page 62 Appendix C EVOS light cubes EVOS ™ light cubes (continued) Light Cube Dyes ™ ™ ™ ™ RFP, Alexa Fluor 546, Alexa Fluor 555, Alexa Fluor 568, Cy3 ™ MitoTracker Orange, Rhodamine Red, DsRed ™ ™ ™ ™ Texas Red Texas Red , Alexa Fluor 568, Alexa Fluor...
-
Page 63: Appendix D Countess ™ 3 Fl Automated Cell Counter And Accessories
™ Countess 3 FL Automated Cell Counter and accessories ™ The following Countess 3 FL 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 FL Automated Cell 1 each AMQAF2000... - Page 64 Appendix D Countess ™ 3 FL Automated Cell Counter and accessories Accessory products (continued) Product Quantity Cat. No. Countess ™ Test Beads (1 × 10 beads/mL) 1 mL C10284 Trypan blue stain (0.4 %) 2 × 1 mL T10282 ™ Each box of Countess Cell Counting Chamber Slides contains 50 slides and 2 ×...
-
Page 65: Appendix E 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 66 Appendix E CSV file format definition Overview (continued) Category Column Name Description Cube 1 concentration Concentration of cells showing fluorescence in the first cube position Cube 1 (%) Percentage of the total cells in brightfield that show fluorescence in the first cube position Cube 1 cells counted Total number of cells counted in the first cube position...
- Page 67 Appendix E CSV file format definition Overview (continued) Category Column Name Description Live circularity min "Live" adjustment slider value for minimum circularity Live circularity max "Live" adjustment slider value for minimum circularity Dead size min Minimum size of "dead" cells in microns Dead size max Maximum size of "dead"...
- Page 68 Appendix E CSV file format definition Overview (continued) Category Column Name Description Cube 2 brightness Second (bottom) light cube adjustment slider value Fluorescence for maximum brightness Template name Template used for count General Software revision Current software version used for count Countess ™...
-
Page 69: Appendix F 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 70: Chemical Safety
Appendix F 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: ·... -
Page 71: Biological Hazard Safety
Appendix F 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 72: Appendix G 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 74 Countess 3 FL Automated Cell Counter_UG_MAN0019567-v3-GUID-2A84EC4A-BB10-453C- A0AD-5AA09BBA4C09-2021/01/28 19:40:13 en 20:14:40.199Z thermofisher.com/support | thermofisher.com/askaquestion thermofisher.com 28 January 2021...