Thermo Scientific Invitrogen Countess II FL User manual

For Research Use Only. Not 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

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.
Revision history: Pub. No. MAN0010644
Revision Date Description
E.0 07 June 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
Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these products, you accept
the terms and conditions of all applicable Limited Use Label Licenses.
TRADEMARKS: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.
©2019 Thermo Fisher Scientific Inc. All rights reserved.

Contents
■CHAPTER 1 About this guide ............................................ 7
Audience ....................................................................... 7
User documentation ............................................................. 7
Text and keyboard conventions .................................................... 7
User attention words ............................................................ 8
Safety alert words ............................................................... 8
■CHAPTER 2 Product information ....................................... 9
Product contents ................................................................ 9
Upon receiving the instrument ................................................ 9
Register your instrument .................................................... 9
Product description ............................................................ 10
Countess™II FL automated cell counter ...................................... 10
Instrument exterior 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
Slide operation ................................................................ 20
Countess™cell counting chamber slide ....................................... 20
Countess™II FL reusable slide .............................................. 21
Countess
™
II FL automated cell counter User Guide
3

■CHAPTER 4 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
■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
■CHAPTER 6 Dilution calculator ........................................ 41
Calculate dilution .............................................................. 41
Dilution calculator ......................................................... 41
Calculate dilution .......................................................... 41
Contents
4
Countess
™
II FL automated cell counter User Guide

■CHAPTER 7 Save results ............................................... 44
Save count results ............................................................. 44
Save screen ............................................................... 44
Save procedure ............................................................ 44
Report ........................................................................ 47
Report file ................................................................ 47
■CHAPTER 8 Instrument settings ...................................... 50
Overview ...................................................................... 50
Instrument settings screen ................................................. 50
Software update ............................................................... 50
Guidelines for software update .............................................. 50
Update the Countess™II/II FL software ....................................... 51
Date/Time ..................................................................... 52
Set the date and time ....................................................... 52
Change light cube .............................................................. 54
Install or change EVOS™light cube ........................................... 54
■CHAPTER 9 Maintenance .............................................. 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 characteristics .................................................... 63
Technical specifications .................................................... 63
Optics .................................................................... 63
Analysis slide ............................................................. 63
EVOS™light cubes .............................................................. 64
LED illumination ........................................................... 64
EVOS™light cubes ......................................................... 64
Contents
Countess
™
II FL automated cell counter User Guide
5

■APPENDIX C Ordering information .................................... 66
Countess™II FL automated cell counter and accessories ............................ 66
Accessory products ............................................................ 66
■APPENDIX D CSV file format ........................................... 68
CSV file format, explained ....................................................... 68
Overview ................................................................. 68
■APPENDIX E Safety ..................................................... 71
Safety alert words .............................................................. 71
Electrical symbols ............................................................. 72
Safety symbols ................................................................ 72
Environmental symbols ......................................................... 73
Safety labels on instruments .................................................... 74
General instrument 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
■Documentation and support ............................................. 80
Customer and technical support ................................................. 80
Limited product warranty ....................................................... 80
Contents
6
Countess
™
II FL automated cell counter User Guide

About this guide
Audience
This user guide is for laboratory sta operating, maintaining, and analyzing data
using the Countess™ II FL Automated Cell Counter.
User documentation
The guides listed below are available for the Countess™ II FL Automated Cell
Counter.
Guide Pub. No.
Countess
™
II FL Automated Cell Counters User Guide MAN0010644
Countess
™
II and Countess
™
II FL Automated Cell Counters Quick
Reference Card (QRC)
MAN0010826
Additional resources are available on the Countess™ Technical Resources page. Go to
www.thermosher.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 example:
Press More.
►Right arrow symbol (►) indicates a menu choice, and separates
successive commands you execute or select from a drop-down or
shortcut menu. For example:
Select More ►Adjust.
1
Countess
™
II FL automated cell counter User Guide
7

User attention words
Two user aention words appear this document. Each word implies a particular level
of observation or action as described below.
Note: Provides information that may be of interest or help but is not critical to the use
of the product.
IMPORTANT! Provides information that is necessary for proper instrument
operation, accurate installation, or safe use of a chemical.
Safety alert words
Four safety alert words appear in this document at points where you need to be aware
of relevant hazards. Each alert word—IMPORTANT, CAUTION, WARNING,
DANGER—implies a particular level of observation or action, as dened 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 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, if not avoided,
will result in death or serious injury. This signal word is to be limited to the
most extreme situations.
Except for IMPORTANT! safety alerts, each safety alert word in this document
appears with an open triangle gure that contains a hazard symbol. These hazard
symbols are identical to the hazard symbols that are axed to the instruments (see
Safety symbols in Appendix E).
Chapter 1 About this guide
User attention words
1
8
Countess
™
II FL automated cell counter User Guide

Product information
Product contents
The Countess™ II FL Automated Cell Counter is shipped with the following
components.
Component Quantity
Countess™II FL Automated Cell Counter (Cat. No. AMQAF1000) 1 each
Power Cord with 4 adaptor cords
(for U.S./Canada/Taiwan/Japan, Europe, or UK)
1 each
Countess™Cell Counting Chamber Slides (50 slides/box) 1 box
Countess™II FL Disposable Slide Holder 1 each
Countess™II FL Reusable Slide Holder 1 each
Countess™II FL Light Cube Removal Tool 1 each
Countess™II USB drive 1 each
Countess™II FL Automated Cell Counter Quick Reference Card 1 each
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 led with the carrier; the warranty does not cover
in-transit damage.
See “Installation“ on page 12 for instructions on installing the instrument.
Visit www.thermosher.com/registercountess to register your instrument. You will
be asked to supply the serial number, your name, and your contact details.
Registering your instrument ensures that you will receive notications of software
upgrades and information on new assays for use with the Countess™ II FL Automated
Cell Counter.
2
Upon receiving the
instrument
Register your
instrument
Countess
™
II FL automated cell counter User Guide
9

Product description
The Countess™ II 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 uorescently labeled cells or trypan
blue stained samples in suspension.
• The Countess™ II FL Automated Cell Counter oers an intuitive user interface,
and provides the option to save data and generate a report, which can then be
transferred to a PC using the USB drive supplied with the instrument or available
separately.
• The cells to be counted are loaded into the instrument either in disposable
Countess™ Cell Counting Chamber Slides or in glass Countess™ II FL Reusable
Slides (“Prepare sample“ on page 19). Each chamber slide contains two enclosed
chambers to hold the sample to allow you to measure two dierent samples or
perform replicates of the same sample.
• The instrument takes 10 seconds per sample for a typical cell count in the
brighteld 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 brighteld channel, the Countess™ II FL Automated Cell
Counter can accommodate two interchangeable EVOS™ uorescent light cubes
(“EVOS™ light cubes“ on page 64), enabling it to be used for multiple-
uorescence research applications.
• When equipped with EVOS™ light cubes, the Countess™ II FL Automated Cell
Counter can be used to perform uorescence assays for cells in suspension,
including simultaneous counts of cells stained with two dierent uorescent
dyes, GFP and RFP expression, apoptosis, and cell viability (live, dead, and total
cells). These assays are compatible with a wide variety of eukaryotic cells.
Countess™II FL
automated cell
counter
Chapter 2 Product information
Product description
2
10
Countess
™
II FL automated cell counter User Guide

Instrument exterior components
1Touch-screen display: The 7‑inch capacitive touch-screen display is the main user
interface of the Countess™II FL Automated Cell Counter. It contains the buttons for all
instrument functions and displays data from the cell count.
2USB ports: The USB ports allow you to transfer and save the cell count data and image to
an external computer for record keeping and printing purposes. You can use the USB drive
supplied with the instrument or any other standard, FAT32-formatted USB drive for data
transfer. If desired, you can plug in a USB mouse into the rear USB port for instrument
control.
Note: The USB ports located in the front and the back of the instrument function the
same. However, the first USB drive connected will be the preferred saving location and
both USB drives cannot be accessed at the same time.
3Slide port: The slide port is used to insert the analysis slide containing the sample into the
counter.
The Countess™II FL instrument accepts both the disposable Countess™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.
4Back 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 the instrument by two captive ¼-turn
fasteners.
5Power switch: The ON/OFF rocker switch is the main power switch. It is not necessary to
use the power switch for day-to-day operation of the instrument.
6EVOS™light cubes: The EVOS™light cubes allow the Countess™II FL Automated Cell
Counter to analyze fluorescently labeled samples. The Countess™II FL Automated Cell
Counter can accommodate two fluorescent light cubes. For more information, see “EVOS™
light cubes“ on page 64.
7USB ports: See 2 above.
8Power input jack: The power input jack connects the instrument to an electrical outlet
through the supplied power cord and the appropriate plug, based on the electrical outlet
configuration in your country.
Chapter 2 Product information
Instrument exterior components
2
Countess
™
II FL automated cell counter User Guide
11

Getting started
Installation
• 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.
• Set up the instrument away from direct light sources, such as windows. Ambient
room lighting can enter the imaging path and aect the image quality.
• Operating temperature range: 4°–32℃ (40°–90°F).
• Relative humidity range: <80%.
IMPORTANT! Do not position the instrument so that it is dicult to turn o 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 at, level, dry surface.
2. Remove the thin plastic protector lm from the touch-screen display.
3. Plug one end of the power cord appropriate for your region into the instrument.
4. Plug the power cord into the electrical outlet. Be sure to use only the power cord
supplied with your instrument. Powering the instrument with an unapproved
power cord may damage the instrument.
3
Operating
environment
Install the
instrument
12
Countess
™
II FL automated cell counter User Guide

1. Turn on the instrument by ipping the power switch on the back of the
instrument (“Instrument exterior components“ on page 11) to the ON position.
The instrument initializes and displays the Home screen.
2. From the Home screen, you can proceed immediately to the assays by inserting a
slide (Chapter 4, “Cell count and cell viability assays“).
Alternatively, you can change or add a prole (Step 3 on page 13) or change
instrument seings (Step 4 on page 13).
3. To change the current prole or to add a new prole to the instrument, press the
Proles buon in the upper left corner.
Proles allow you to create customized count preferences (i.e., gate counts based
on cell size, brightness, circularity, and/or relative uorescence intensity) (“Load
prole“ on page 14).
4. To change instruments seings, press the Instrument Seings buon in the
upper right corner.
Instrument seings allow you to update the Countess™ II software, change the
date and time, and install or change up to two EVOS™ light cubes (Chapter 8,
“Instrument seings“).
Turn ON the
instrument
Chapter 3 Getting started
Installation
3
Countess
™
II FL automated cell counter User Guide
13

Load profile
Proles screen allows you to create and save up to 9 customized proles. Each custom
prole denes the count parameters (size, brightness, circularity, and uorescence
intensity) and automatic instrument functions (Auto Lighting and Auto FL
Threshold) for a consistent and streamlined workow.
• You can access the Proles screen from the Home, Capture, Results, Advanced™,
or Adjust screens.
• The current prole is displayed on the upper left corner of the Home, Capture,
Results, Advanced™, or Adjust screens.
• Automatic instrument functions (below) and count parameters (“Count
parameters“ on page 15) are dened in the Edit prole screen (see “Add/edit a
prole“ on page 16).
• The Default prole contains default count seings and cannot be edited.
• The count parameters specied in the selected prole are applied to all new cell
counts.
• If you have already performed a count, loading a new prole 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 seing that is saved as part of the protocol (size, brightness, or
circularity) on the Results screen, the prole name is appended with the (*)
symbol.
You can turn the Auto FL Threshold and Auto Lighting functions ON and OFF using
the Auto FL Threshold and Auto Lighting checkboxes in the Edit prole screen
(“Add/edit a prole“ on page 16).
•Auto FL Threshold: Automatically applies threshold in uorescence channels to
subtract background uorescence for improved analysis despite variable
background levels between samples. This function is available only for
instruments equipped with the optional EVOS™ light cubes.
•Auto Lighting: Automatically illuminates the sample in the brighteld for
increased sample-to-sample consistency and decreased user-to-user variability.
Profiles screen
Automatic
instrument
functions
Chapter 3 Getting started
Load profile
3
14
Countess
™
II FL automated cell counter User Guide

Count parameters are adjusted in the Edit prole screen using the parameter sliders.
Parameter sliders correspond to a single channel, which is selected using the channel
selection radio buons located above the sliders.
•Size: As you move the slider up, the algorithm includes larger objects in the
count. As you move the slider down, only the smaller objects are counted.
•Brightness: As you move the slider up, the algorithm includes the brighter
objects in the count. As you move the slider down, only the dimmest of objects
are counted.
•Circularity: As you move the slider up, the algorithm includes more objects with
shapes other than circular in the count. As you move the slider down, only the
objects that are perfect circles are counted.
•Fluorescence intensity: As you move the slider up, the algorithm includes the
objects that uoresce more brightly in the count. As you move the slider down,
only the dimmest of objects are counted.
•Size, brightness, and uorescence 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 adjust only the upper or the lower boundary, move the upper or the lower
handle in the desired direction. This will also change the range of values within
which the cells are counted.
• The circularity slider only sets a single threshold value; cells that fall below the
set value are counted, and cells that are beyond this range are excluded.
To adjust the threshold for circularity, drag the slider in the desired direction.
Count parameters
Chapter 3 Getting started
Load profile
3
Countess
™
II FL automated cell counter User Guide
15

1. Press the Proles buon located on the upper left corner of the screen to open
the Proles screen.
2. Press the desired prole to select, and then press Load.
The instrument will load the count parameters specied in the selected prole
and return to the previous screen.
3. To return to the previous screen without loading the new prole, press the
previous buon.
The instrument will keep the saved prole, but return to the previous screen
without loading it.
1. Press the Proles buon located on the upper left corner of the screen to go to
the Proles screen.
2. To add or edit a new prole, select an empty or an existing prole, and then
press Edit. The Edit screen for the selected prole opens.
Note: The Default prole contains default count seings 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 function
ON or OFF (“Automatic instrument functions“ on page 14).
Define count parameters for the brightfield channel:
To dene the new count parameters in the brighteld channel:
1. From the Trypan Blue selection box, select the Live or Dead radio buon.
2. Adjust the size, brightness, and circularity thresholds using the corresponding
parameter slider (“Count parameters“ on page 15).
Load a profile
Add/edit a profile
Chapter 3 Getting started
Load profile
3
16
Countess
™
II FL automated cell counter User Guide

Define count parameters for the fluorescent channels
1. To dene the new count parameters for uorescence assays (available only for
instruments equipped with the optional EVOS™ light cubes):
a. From the FL selection box, select the desired channel using the
corresponding radio buon.
The available options are BF (brighteld) and up to two uorescence
channels, depending on the light cubes installed (GFP and TxRed in the
following example).
b. Dene the new count parameters for the selected channel using the
corresponding parameter slider (“Count parameters“ 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 uorescence channel allows you to gate
count results based on relative uorescence intensity in that channel.
c. Repeat for the remaining channels, as necessary.
Note: You can further adjust the size, brightness, or circularity parameters
for the selected prole before or after performing a cell count, as needed.
You can then save these additional changes to the count parameters to the
current prole or as a separate prole directly from the Advanced™ screen
(see “Save as new protocol“ on page 29)
Chapter 3 Getting started
Load profile
3
Countess
™
II FL automated cell counter User Guide
17

2. To assign a name to the new prole or to change the name of the existing prole,
press the Prole name text box. The alpha-numeric keypad opens.
3. Type in the desired prole name using the alpha-numeric keypad.
To enter symbols, press the symbol (@%&) key. To return to the alpha-numeric
keypad, press ABC.
4. Press Enter to save the name and return to the Edit prole screen.
To return to the Edit prole screen without saving the name, press the close
buon.
5. Press Save to save the new prole, and then press Close in the conrmation
screen to return to the Proles screen.
To return to the Proles screen without saving, press Cancel.
6. On the Proles screen, press Load. The instrument will load the count
parameters specied in the selected prole and return to the previous screen.
7. To return to the previous screen without loading the new prole, press the
previous buon. The instrument will keep the saved prole, but return to the
previous screen without loading it.
• The maximum display length for prole names on the screen is seven characters.
• If the prole name is over seven display characters, the name is shortened to the
rst seven characters with “…” at the end. For example, “MyProle” is shortened
to “MyPro…”.
• If you change any seing that is saved as part of the protocol (size, brightness, or
circularity) on the Results screen, the prole name is appended with the “(*)”
symbol.
For example, “Count” becomes “Count (*)”.
• When a prole with name of over four or ve display characters is edited, only
the rst four or ve characters is displayed and the name is appended with “…
(*)”.
For example, “MyProle” becomes “MyPro…(*)”.
Display of profile
names
Chapter 3 Getting started
Load profile
3
18
Countess
™
II FL automated cell counter User Guide

Prepare sample
To obtain the best results, follow these recommendations:
• Ensure that the cell sample is homogeneously mixed.
• The measurement range extends from 1 × 104–1 × 107 cells/mL, but the optimal
range is 1 × 105–4 × 106 cells/mL.
• For accurate results in cell viability assays, ensure that the counting area is
covered with the cell suspension and count the cells immediately after staining
per the assay protocol.
• Do not press the optical surfaces of the chamber slides. Hold the slides by the
edges.
• Take care to avoid forming bubbles in the sample.
• Sterile ltering and centrifugation can be used to remove precipitates common
within trypan blue solutions. Alternatively, avoid mixing and vortexing trypan
blue stock solutions to allow precipitates to remain at the boom of tube, thereby
promoting more accurate cell counts. Also, precipitates 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 pipeing 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 sele in the chamber for 30 seconds, and then insert the
slide into the slide port (“Instrument exterior components“ on page 11). You will
hear a soft click, if the slide is pushed in correctly.
4. To remove the slide, push the slide gently into the instrument until it “clicks”
and a spring pushes the slide out. Grasp the slide and pull it out the rest of the
way.
Note: After using the Countess™ Cell Counting Chamber Slides, appropriately
dispose of them as biohazardous waste. Do not reuse the disposable chamber
slides.
Recommendations
Load Countess™
chamber slide
Chapter 3 Getting started
Prepare sample
3
Countess
™
II FL automated cell counter User Guide
19

1. Before loading your sample into the Countess™ II FL Reusable Slide, place a
cover slip on the counting chamber, making sure the cover slip is clean and free
of grease.
2. 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 lm of uid under the cover slip.
3. After using the Countess™ II FL Reusable Slide, rinse the glass slide and cover
slip with water, and then clean with 70% ethanol. Use Kimwipes™ laboratory
tissues to clean and dry the slides, as needed.
Note: Each chamber in the Countess™ Cell Counting Chamber Slide or the
Countess™ II FL Reusable Slide has a 10-µL sample capacity. Do not overll 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-
specic carriers.
1. To use the plastic, disposable Countess™ Cell Counting Chamber Slide with the
Countess™ II FL Automated Cell Counter, insert the slide carrier (black, see
image) into the slide port of the instrument until it clicks into place.
Note: The Countess™ II FL Automated Cell Counter is shipped with the
disposable slide carrier already installed
Load Countess™II
FL reusable slide
Countess™cell
counting chamber
slide
Chapter 3 Getting started
Slide operation
3
20
Countess
™
II FL automated cell counter User Guide