Countess™ Automated
Cell Counter
Catalog no. C10227
Rev. Date: 15 September 2009
MP10227
User Manual
ii
Table of Contents
Product Contents......................................................................................................... iv
Safety Information ........................................................................................................v
Product Specifications ............................................................................................... vii
Description of Countess
Introduction.......................................................................................... 1
Product Overview.........................................................................................................1
Methods................................................................................................3
Getting Started...............................................................................................................3
General Guidelines .......................................................................................................9
Using the Countess
Cleaning and Maintenance........................................................................................19
Updating and Calibrating the Countess
Troubleshooting ..........................................................................................................22
Countess
™
Automated Cell Counter Error Codes .................................................24
™
Automated Cell Counter ...........................................viii
™
Automated Cell Counter .....................................................10
™
Automated Cell Counter..................20
Appendix ............................................................................................ 25
Accessory Products.....................................................................................................25
Technical Support .......................................................................................................26
Purchaser Notification................................................................................................27
iii
Product Contents
Countess™
Automated Cell
Counter
Upon
Receiving the
Instrument
The contents of the Countess™ automated cell counter are listed
below:
Component Quantity
™
Countess
Power Cord with 4 adaptor cords 1
(for U.S./Canada/Taiwan/Japan, Europe, or UK)
Countess
Trypan blue stain (0.4%) 2 × 1 mL
Countess
Quick reference card (QRC) 1
The complete instruction manual is available on our website at
www.invitrogen.com/countess.
See page vii for specifications and description of the Countess™
automated cell counter.
Examine the instrument carefully for any damage incurred
during transit. Ensure that all parts of the instrument including
accessories listed above are included with the product. Any
damage claims must be filed with the carrier. The warranty does
not cover in-transit damage.
See page 3 to install the instrument.
Upon receipt, store trypan blue stain at room temperature.
automated cell counter 1
™
cell counting chamber slides 1 box of 50
™
USB drive 1
iv
Safety Information
y
Safety
Precautions
Review and follow the safety instructions below.
Do not install the instrument in heavy humidity such as a
greenhouse or an incubator to avoid a danger of electric
shock. If water or other material enters the instrument, the
adaptor, or power inlet, disconnect the power cord and
contact a service person. For operating environment, refer to
Product Specifications (page vii).
Do not touch th
Always ensure that the power supply input voltage matches
the voltage available in your location.
This instrument is air-cooled so its surfaces may become hot
during operation. When installing the instrument, leave a
space of more than 10 cm (4 inches) around it and do not
place any objects between the instrument and the wall.
Do not install the instrument on a slant or a place prone to
vibrations, which induces the risk of instrument malfunction
or damage of the instrument.
Never insert an
this could result in electrical shock, personal injury, and
equipment damage.
Plug the power cord firmly into the wall outlet and AC
adapter.
To avoid potential shock hazard, make 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
USB drive).
e main plug or power cord with wet hands.
objects into the air vents of the instrument as
Continued on next page
v
Safety Information, Continued
Safety
Precautions,
continued
Symbols
Do not use metal objects when assembling or dissembling a
battery.
Do not heat the battery or expose it to heat. For operating
environment, see Product Specifications (page vii).
If the instrument emits smoke, disconnect the power cord
from the wall outlet and contact a service person.
The symbols used on Countess™ automated cell counter are
explained below.
Used on the instrument to indicate a warning. Consult the
manual to avoid possible personal injury or instrument damage.
Protective earth (Ground)
WEEE (Waste Electrical and Electronic Equipment) symbol
indicates that this product should not be disposed of in unsorted
municipal waste. Follow local municipal waste ordinances for
proper disposal provisions to reduce the environmental impact
of WEEE. Visit www.invitrogen.com/weee for collection and
recycling options.
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 specified by Invitrogen.
This product conforms to UL61010-1/CSA C22.2 No. 61010-1
“Safety Requirements for Electrical Equipment for Measurement,
Control, and Laboratory Use, Part I: General Requirements.”
Instruments bearing the TUV symbol are certified by TUV
Product Services to be in conformance with the applicable safety
standards for the US and Canada.
vi
Product Specifications
Countess™
Automated Cell
Counter
Specifications
Environmental Conditions
Operating Power 100–240 VAC, 1.5 A
Frequency 50/60 Hz
Electrical input 12 VDC, 1.0 A
Installation site: Indoor use only
Operating Temperature: 10–35°C
Maximum Relative Humidity: 20–80%
Altitude: <2,000 m
Transient Category: Installation categories II
Pollution Degree: 2
Degree of Protection: IPX0
Instrument Specifications
Instrument Type: Benchtop cell counter
Processing Time: <1 minute
Cell Sample Range: 1 × 10
Software: Countess
www.invitrogen.com/countess
Instrument Dimensions: 27 cm (w) × 20 cm (d) ×
19 cm (h)
Weight: 2.1 kg (4.6 lbs)
The Countess
(2.3X objective and 3.1 Mega pixel).
Countess
Material: Polymethyl methacrylate
(PMMA)
Dimensions: 75 mm (w) × 25 mm (d) ×
1.8 mm (h)
Chamber Depth: 100 μm
Chamber Volume: 10 μL
Countess™ USB Drive: 1 Gigabyte
™
automated cell counter is equipped with a camera
™
Cell Counting Chamber Slide Specifications
4
–1 × 107
™
Software
vii
Description of Countess™ Automated Cell
Counter
Front View of
Countess
™
Automated
Cell Counter
The Countess™ automated cell counter is a benchtop automated
cell counter that performs cell count and viability measurements
using trypan blue stain.
The front view showing various parts of the Countess
automated cell counter is shown below.
Power button
The Power Button is used to turn the instrument on and off. The
red status light indicates that the instrument is off; the green
status light indicates that the instrument is on.
Touchscreen display located in the front of the instrument
contains buttons for all the functions needed and displays data
from the cell count.
Slide port
™
The Slide port is used to insert the Countess
chamber slide containing the sample with trypan blue into the
counter for analysis.
USB port
The USB port allows you to transfer and save the cell count data
and image to your computer for record keeping and printing
purposes. The USB drive supplied with the instrument or any
other standard USB drive is inserted into the USB port for data
transfer. See page 16 for Transferring Data to a PC.
cell counting
™
Slide
Port
USB port
Touchscreen
viii
Power
button
Continued on next page
Description of Countess™ Automated Cell
Counter,
Rear and Side
View of
Countess
Automated
Cell Counter
™
Rear View Side View
Continued
The rear and side view showing various parts of the Countess™
automated cell counter.
Power Inlet
Connect the counter to an electrical outlet using the supplied
power cord and the appropriate plug, based on the electrical outlet
configuration in your country.
Image Adjustment (Focus) Knob
The Image Adjustment (Focus) knob is used to adjust the image
quality to obtain better contrast between live (bright centers) and
dead (dark blue centers) cells. This is important to obtain accurate
cell counts and viability measurements. See page 14 for details.
Foc
us
Lock Knob
The Focus lock knob is used to lock the Image Adjustment (Focus)
knob once the image is optimized. There is no need to use the
Focus lock knob, but is available for convenience, if you are
measuring multiple samples of the same cell type.
Power
inlet
Focus
knob
Focus
Continued on next page
lock
ix
Description of Countess™ Automated Cell
Counter,
User Interface
Countess™
Cell Counting
Chamber Slide
Continued
The touch screen user interface of the Countess™ automated cell
counter is used to operate the instrument and consists of:
The touch screen buttons to operate the instrument.
The Digital Display shows the image of cells and sample
data.
Touch
Screen
Button
The Countess™ cell counting chamber slides are plastic, disposable
enclosed chambers that hold the sample in two separate chambers
(A and B) for replicates. The cell counting occurs in the central
location of the counting chamber. The entire volume of cells
counted is 0.4 μL, the same as counting four (1 mm × 1 mm
squares in a standard hemocytometer.
)
x
Introduction
Product Overview
Countess™
Automated
Cell Counter
The Countess™ automated cell counter uses state-of-the-art optics
and image analysis to automate cell counting. The Countess
automated cell counter is a benchtop counter designed to measure
cell count and viability (live, dead, and total cells) accurately and
precisely, using the standard trypan blue technique.
Using the same amount of sample that you currently use with the
hemocytometer, the Countess
than a minute per sample for a typical cell count and is compatible
with a wide variety of eukaryotic cells and provides information
on cell size.
The Countess
interface, and provides the option to save and print cell count data
using the Countess™ software (download from
www.invitrogen.com/countess) and USB drive supplied with the
instrument or available separately.
The Countess
Countess
chambers to hold the sample to allow you to measure two different
samples or perform replicates of the same sample. The cell
counting occurs in the central location of the counting chamber
and the volume counted is 0.4 μL, the same as counting four
(1 mm × 1 mm) squares in a standard hemocytometer.
See page viii for details on various parts of the counter.
™
cell counting chamber slides that contain two enclosed
™
™
automated cell counter takes less
™
automated cell counter offers an intuitive user
™
automated cell counter is supplied with disposable
Continued on next page
1
Product Overview, Continued
p
System
Overview
Features
The Countess™ automated cell counter performs viability and cell
counting measurements using the trypan blue method of dead-cell
staining combined with advanced image analysis.
Cell sample is mixed with trypan blue and loaded into a Countess™
cell counting chamber slide. The camera acquires cell images from
the sample on the slide and the image analysis software
automatically analyzes acquired cell images, and measures cell
count and viability using the trypan blue stain.
A single, sample measurement within a minute provides the
following data:
Live and dead cell concentration/mL
Total cell concentration/mL
Viability (% live cells to total cells)
Mean diameter
Cell images
Graphical data representation
Important features of the Countess™ automatic cell counter are:
User-friendly, benchtop design for simple, fast, automated cell
count and viability measurements within a minute
Provides data on cell size and is compatible with a wide variety
of eukaryotic cells without the need for any special changes
between large or small sizes
Measures cell concentrations ranging from 1 × 10
cells/mL and cells with sizes ranging from 5 μm to 60 μm
Uses dis
osable counting chamber slides that eliminate washing
steps and cross contamination between samples
Saves and prints cell count data including images using the
Countess
™
software and Countess™ USB drive
Presents comprehensive data with graphical reports and as a
.CSV (comma separated value) file for sample comparisons
Allows you to gate and analyze cells based on cell size
4
to 1 × 107
2
Getting Started
Installing the
Countess
Automated
Cell Counter
1. After unpacking the instrument, place the instrument on a flat,
™
level, dry surface.
2. Plug one end of the supplied power cord into the Countess
instrument. To the other end attach the appropriate plug
adaptor, based on the electrical outlet configuration in your
country.
3. 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.
To register your Countess
www.invitrogen.com/countess. Enter the instrument serial
number, your name, and your contact details. Registering your
instrument ensures that you receive notifications of software
upgrades and fast service.
4. When you are ready to use, start the Countess
pressing the Power button.
Methods
™
automated cell counter, visit
™
instrument by
Continued on next page
™
3
Getting Started, Continued
p
Optional:
Installing the
Countess
Software
Start-Up
Screen
The Countess™ instrument is designed for stand-alone use and does
not require the use of an external computer.
™
If you wish to archive data and generate reports, you must transfer
data to your computer, and use the Countess
and
rint reports (see instructions, below). Alternatively, data stored
in the .CSV file may be transferred to your computer by the USB
drive and imported into any spreadsheet program, without the need
for Countess
™
software.
Computer requirements (Countess
Macintosh Operating systems)
USB port (1.1 or later, 2.0 is recommended)
Windows XP/2000/Vista
1. Visit www.invitrogen.com/countess and follow instructions
on the page to install the Countess
2. When the software is installed, the Countess
counting software icon appears on your desktop.
3. To download data, use any USB drive to capture data from
your experiment and transfer data to your computer as
described in Transferring Data to a PC (page 16).
When the instrument is turned on, the Start Up screen is
displayed. Here you can proceed immediately to cell counting
(page 9), set up the instrument for cell or bead
screen brightness.
Start Up Screen
™
software to generate
™
is not compatible with
™
software.
™
automated cell
count, or adjust the
Screen Brightness
Use the scroll buttons to adjust the screen brightness.
Continued on next page
4
Getting Started, Continued
Settings
Screen
Cell Mode
Parameters
Screen
Press Settings from the Start-Up screen to display Settings.
The Settings menu allows you to set up the following:
Parameters (see below and next page for details)
Calibration to calibrate the instrument (page 20)
Update to install new firm
available
Date and Time to set up date and time (page 8)
Count mode to operate the ins
(choose Cells) or bead counting (choose Beads)
Press Parameters from the Settings screen to display Cell Mode
Parameters Screen.
The Countess
the majority of cultured cell types which can be accurately counted
without changing any of the default parameters. The Parameters
function allows you to change the image analysis algorithm for
specific or mixed cell types, and the specific parameters must be
determined empirically.
™
instrument comes with pre-set parameters to match
ware versions as they become
trument for cell counting
Continued on next page
5
Getting Started, Continued
Cell Mode
Parameters
Screen,
continued
The Cell Mode Parameters are described below:
Sensitivity (refers to the contrast of the objects from the
background). Adjusting the sensitivity higher makes the
instrument more sensitive to objects; useful for cells that do not
stain well with trypan blue while adjusting the sensitivity
lower makes the instrument less sensitive and is useful if there
is a lot of background.
Minimum cell size is used to select the lowest particle size to
include in the count. The size is set in 1 micrometer
increments. The lower range of the Countess
5 μm. However, you can select a minimum cell size as low as
2 μm; when a cell size outside of the usual range is selected, the
numbers indicating particle size are shown in red.
Maximum cell size is used to select the maximum particle size
to include in the count. The size is set in 1 micrometer
increments. The maximum size that can be set is 60 μm.
Circularity is used to determine the objects to include in the
measurement based on roundness. Increasing the value from
80% requires objects to be more round for inclusion in the
measurement. Decreasing the value from 80% allows objects to
be less round. Adjusting this may be useful if the cell type is
not particularly circular or perhaps oddly shaped.
After modifying any parameters:
Press Apply to make the changes or,
Press Default to restore default parameters or,
Press Save to create a protocol. Save the protocol name and
user name in the appropriate fields, and press Save again. The
protocol is now saved and appears highlighted in the protocol
menu. To use the highlighted protocol, press Apply.
™
instrument is
Continued on next page
6
Getting Started, Continued
Cell Mode
Parameters
Screen,
continued
Note: Once a protocol is saved, it is available for use at any time. To access
saved protocols, press Settings > Parameters > Load Protocol. Use the up
and down arrows to find your saved protocol. Once the protocol is located,
use the touch screen to highlight the protocol and press Apply. Once the
protocol of interest is selected, count cells using the standard procedure.
7
Getting Started, Continued
g
Date and
Time Setup
The date and time is already preset when you receive the
instrument. To reset the date and time, follow these steps:
1. Turn on the Countess
Power button. The Start up screen is displayed after a few
seconds.
2. Press Settings and then press Date and Time.
3. The Date/Time Properties screen is displayed. To select the
month and day, scroll to the appropriate month usin
keys and then press the day on the calendar.
Note: Use a pointed object, like a stylus or pipette tip, to push the small
buttons on the calendar.
™
automated cell counter by pressing the
the arrow
4. To select the time, scroll to the appropriate time and select
Automatically adjust clock for daylight saving, if needed.
Press Apply or OK to make the date/time changes. The
updated date/time is displayed on the top of the window.
Once the date/time is set, there is no need to set it each time the
instrument is turned on.
5. Press Close to exit the screen.
8
General Guidelines
Countess™
Cell Counting
Chamber
Slides
N
E
D
M
A
M
T
O
I
O
C
N
E
R
The Countess™ cell counting chamber slides supplied with the
counter or available separately from Invitrogen (page 25) are
specially desig
ned for use exclusively with the Countess
automated cell counter. Use of other slides result in inaccurate cell
counts and can damage the Countess
To obtain the best results, follow these recommendations:
Wear gloves during sample handling.
Do not touch the optical surfaces of the Countess
counting chamber slides. Hold the chamber slides by the
edges.
Use the Countess
temperature only (10–35ºC).
For accurate viability count results, ensure the counting area is
covered with cell suspension and count cells within 3 minutes
of mixing the cells with trypan blue solution as trypan blue is
toxic to cells. For best data with biological samples, we
recommend counting at least two samples and taking an
average.
The Countess
calibrated. To recalibrate your instrument, see page 20.
The Countess
of data. Save your data to the USB drive after each reading.
You may transfer the data to your PC, using the USB drive
immediately as described in Transferring Data to a PC (page
16).
After usi
ng Countess™ cell counting chamber slides,
appropriately dispose slides as biohazardous waste. Do not
reuse the chamber slides.
™
™
instrument.
™
cell
™
automated cell counter at room
™
automated cell counter is supplied pre-
™
automated cell counter memory holds one set
9
Using the Countess™ Automated Cell Counter
Materials
Needed
Cell Counting
Procedure
The Countess
™
automated cell counter is supplied
precalibrated. However, if you wish to calibrate the Countess
automated cell counter, see page 20.
Instructions are provided in this section for preparing the c
sample with trypan blue dye for use with disposable
Countess
count using the Countess
Visit www.invitrogen.com/countess
™
cell counting chamber slides for automated cell
™
counter.
for additional application
notes, technical notes, and a list of validated cell lines
including adherent, primary, and suspension cells that have
been tested using the Countess
™
Automated Cell Counter.
Cell sample
™
Countess
cell counting chamber slides (supplied with the
instrument or available separately, page 25)
Trypan blue stain (0.4%); supp
lied with the instrument or
available separately, page 25
Optional:
Countess
™
USB drive for data transfer, supplied with
the instrument or available separately (page 25)
1. Push the Power button to start the instrument. The Start-up
screen is displayed.
ell
™
10
™
2. The Countess
automated cell counter is preset to Cell mode.
If you have changed the mode to bead for bead counting (page
17) or calibration (page 21), be sure to change the mode to
Cells Mode prior to coun
ting cells by pressing Settings and
then Cells.
Continued on next page
Using the Countess™ Automated Cell
Counter,
Cell Counting
Procedure,
continued
Continued
3. Add 10 μL of your sample to 10 μL supplied trypan blue stain.
Mix gently by pipetting up and down.
4. Add 10 μL of the sample mixture to the chamber ports on one
side of the Countess
chambers of the slide are labeled “A” and “B” for easy tracking
of your samples. You can count the cells from one chamber or
both chambers.
5. Insert the Countess
first into the slide inlet on the instrument, making sure that the
sample side A is inserted completely into the instrument. You
will hear a soft click, if the slide is pushed in correctly. Each
chamber is counted separately.
™
cell counting chamber slide. The two
™
cell counting chamber slide, sample side A
6. Press the Count Cells or Next Sample button.
Continued on next page
11
Using the Countess™ Automated Cell
Counter,
Cell Counting
Procedure,
continued
Continued
7. Adjust the image by pressing the Zoom button. Navigate by
pressing the location you like to see on the grid.
8. While viewing cells in the Zoom mode, use the Focus knob to
adjust the image.
Note: After you have counted the first sample, you may not have to
use the Focus knob again. If measuring multiple samples of the same
approximate size, you may lock the Focus knob. You are able to
unlock the knob later, to adjust the image.
12
Optimize the image for analysis such that:
Live cells have bright centers and dark edges (see image,
next page)
Dead cells have a uniform blue color throughout the cell
with no bright centers, (see image, next page)
Continued on next page
Using the Countess™ Automated Cell
Counter,
Continued
Cell Counting Procedure, continued
Correct Image
Live
Incorrect Image
Live
Dead
Dead
9. When you are satisfied with the image, press Count Cells.
Continued on next page
13
Using the Countess™ Automated Cell
Counter,
Cell Counting
Procedure,
continued
Continued
10. The instrument takes approximately 30 seconds to count each
sample, and the cell count for live, dead, and total cells, and
percentage viability is displayed on the screen. Record the cell
count, or insert a USB drive and press Save (see page 16).
11. After a cell count is performed, use the Details button to see
how each cell was counted. Objects are circled in blue, red, or
black. Blue indicates cells that are counted as live, red indicates
cells that are counted as dead, and black indicates objects that
are excluded from the count.
12. To see more details on the data as well as graphical
representation of the data, press the More Data button.
Note: The data window in More Data allows you to further analyze
acquired data by viewing a specific data set. The data window is set
using particle size that you specify. Once the window is set, the
concentrations and particles counted are recalculated to reflect the data
window you set. Use the arrows to view only the window of processed
data you wish to view. Once you have set the window, press Apply to
view
the information box with the recalculated data set specifically for
the data window.
14
Continued on next page
Using the Countess™ Automated Cell
Counter,
Cell Counting
Procedure,
continued
Continued
After reviewing the data, press the Close button to return to
the main screen.
The Calculator button allows you to quickly calculate
adjustments to the cell suspension to obtain a desired
concentration.
13. To count the cells in the other side of the slide chamber (side
B), remove the Countess
pushing in the slide slightly and then pulling the slide out.
Turn the slide around and reinsert into the slide inlet and
repeat the procedure.
™
slide after side A is counted by
14. The Countess
of data. To save your data for future analysis or archiving, you
must record the data or save after each reading. See page 16
for Transferring Data to a PC.
15. After recording or saving the data, remove and discard the
slide appropriately as biohazardous waste.
16. At this point, the Countess
for another sample. If you are not using the instrument, press
the Power button to turn off the instrument.
Note: If the touch screen is not responding, you can turn off the instrument
by pressing and holding the Power button for 4 seconds.
™
automated cell counter memory holds one set
™
cell counter instrument is ready
Continued on next page
15
Using the Countess™ Automated Cell
Counter,
Optional:
Transferring
Data to a
Computer
Continued
1. To archive your data or generate a printed report, insert
Countess
USB port
2. Save your data on the USB drive by pressing the Save button
on the main screen. The image and the counting data are
saved.
The numerical data is also automatically saved as a .CSV file
that can be opened with any spreadsheet program. To delete
all data from the .CSV file and start with a blank file, press
Start .CSV file button.
3. Enter the file name using the keypad buttons displayed on the
Save menu.
4. Transfer the Countess
You may open the .CSV file using a spreadsheet program. To
see the image and generate a report, use the Countess™
software.
5. Click on the Countess
on your PC (see Installing the Countess
6. Open a file using the Open button. Select the saved file you
wish to open.
7. Zoom in on the image obtained by clicking in the zoom grid,
(this screen also allows you to examine which cells have been
counted as live or dead).
8. Click on Prepare Report to obtain a printable version of the
image and data.
™
USB drive into the USB port.
™
USB drive to the USB port on your PC.
™
automated cell counting software icon
™
Software, page 4).
16
Continued on next page
Using the Countess™ Automated Cell
Counter,
Using the
Beads Mode
for Counting
Beads
Continued
The Countess™ automated cell counter can also be used to count
beads using the Bead mode of the instrument.
1. Push the Power button to start the instrument. The Start-up
screen is displayed.
2. Press Settings and then press Beads to place the instrument
into bead counting mode.
3. Add 10 μL of beads to 10 μL supplied trypan blue stain. Mix
gently by pipetting up and down.
4. Add 10 μL of the sample mixture to the chamber ports on one
side of the Countess
page 10. You can count beads from one chamber or both
chambers.
5. Insert the Countess
into the slide inlet on the instrument as shown on page 11,
ma
the instrument.
6. Press Count Beads button.
7. Adjust the bead image by pressing the Zoom button. Navigate
the fields by pressing the location you like to see on the grid.
Use the focus knob to adjust the image.
king sure that the sample side is inserted completely into
™
cell counting chamber slide as shown on
™
cell counting chamber slide with beads
Continued on next page
17
Using the Countess™ Automated Cell
Counter,
Using the
Beads Mode
for Counting
Beads,
continued
Continued
8. When you are satisfied with the image, press the Count Beads
button.
9. The instrument takes approximately 30 seconds to count each
sample and the bead count is displayed on the screen. Record
the bead count.
10. To count beads in the other side of the slide chamber (side B),
remove the slide after side A is counted, turn the slide around,
and reinsert into the slide inlet to repeat the counting
procedure.
11. After counting beads, place instrument into Cell Count mode
for counting cells by pressing Settings button and then
pressing the Cells button.
18
Cleaning and Maintenance
Cleaning and
Maintenance
Changing
Battery
Clean the surface of the Countess™ instrument with a damp cloth.
To clean the LCD screen, turn off the Countess
disconnect the power cable, and clean the LCD screen with a soft
cloth lightly moistened with LCD cleansing detergent. Cleaning
the screen with excessive force can damage the LCD the screen.
Wipe the screen dry immediately. Do not reuse the counting
chambers.
™
The Countess
automated cell counter does not need regular
maintenance. To troubleshoot problems with Countess
counter, contact Technical Support (page 26). Do not perfor
repairs or service on the Countess
the instrument.
To change the battery:
™
1. Turn off the Countess
automated cell counter and disconnect
the power cord.
2. Open the battery cover at the bottom of the Countess
automated cell counter.
™
instrument,
™
cell
™
instrument to avoid damaging
m any
™
3. Remove the used battery and insert a new battery (battery
type CR2032).
4. Close the battery cover. Reconnect the power cord.
5. Reset the date and time on the counter as described on page 8.
19
Updating and Calibrating the Countess™
p
Automated Cell Counter
Introduction
Updating the
Counter
Instructions are included below to update and calibrate the
Countess
™
automated cell counter to ensure its optimal
performance. The update feature allows you to update the counter
when new firmware versions are available. The calibration function
sets the background of the instrument to a solution of trypan blue
in the slide.
The Countess™ automated cell counter is supplied precalibrated.
However, if you have updated the Countess
™
firmware, or if you
choose to use a different formulation of trypan blue that is not
supplied with the instrument, you will need to calibrate the
Countess
™
automated cell counter.
1. Save any data from the .CSV file you wish to archive as follows:
Go to the save menu>Press View .CSV file>Press Save to USB
2. Start a new .CSV file as follows:
Go to the Save Screen>Press Start new .CSV>When prompted,
press OK to erase previous data and start new .CSV file.
3. Visit the Countess
™
website at www.invitrogen.com/countess
to download the firmware which is supplied as a .zip file to
your desktop.
4. Unzip the .zip file and save the firmware to a USB drive
supplied with the instrument.
5. Insert the USB drive into the USB port on Countess
6. Push the
Settings
7. Press
Power
.
Update
button to start the instrument and press
.
™
counter.
8. The instrument prompts you to update, press OK. The update
process takes a few minutes.
9. When the update is complete, you are prompted to restart the
instrument by pressing
10. Once the start u
Restart
.
screen is displayed, calibrate the instrument as
described on the next page.
20
Continued on next page
Updating and Calibrating the Countess™
Automated Cell Counter,
Calibrating
the Counter
1. Push the Power button to start the instrument. The Start-up
screen is displayed.
2. Press Settings and then press Calibration.
3. To recalibrate the Countess
solution with 10 μL of a standard buffer, such as phosphate
buffered saline (PBS). Mix thoroughly.
Continued
™
Counter, mix 10 μL trypan blue
4. Add 10 μL of the sample mixture to the chamber ports on one
side of the Countess
5. The instrument takes approximately a minute to calibrate.
6. After calibration is complete, press the Re-start button to restart
the instrument, and proceed to cell counting.
There is no need to recalibrate each time the instrument is turned on.
™
cell counting slide. Press Start.
21
Troubleshooting
Problem Cause Solution
Inaccurate cell
count
Sample handling
Do not insert the Countess
counting chamber slide upside-down
as this may introduce liquid into the
instrument that could damage it.
Do not reuse the Countess
counting chamber slides, as leftover
dye from the previous reading may
affect the next reading.
Do not use any other counting
chambers such as a glass
hemocytometer with the Countess™
counter as it results in inaccurate cell
count and may damage the
instrument.
Ensure that the sample covers the
entire counting area and the
Countess™ cell counting chamber
slide is inserted completely into the
counter.
Low and high
readings
The Countess™ automated cell counter is
designed to read samples from 1 × 10
cells/mL to 1 × 10
highest accuracy between 1 × 10
and 4 × 10
6
7
cells/mL.
If your sample is not in this range, you
may need to dilute the sample or add
more cells and read the sample again.
Poor image
quality
While viewing cells in the Zoom mode, use
the focus knob to adjust the image to
ensure that live cells have bright centers,
and dead cells have dark/blue centers.
Cell clumping Ensure the cells are not clumped.
--
To maintain instrument sensitivity, we
recommend that you calibrate the counter
each year as described on page 21.
™
cell
™
cell
cells/mL, with the
5
cells/mL
4
22
Continued on next page
Troubleshooting, Continued
Problem Cause Solution
Error codes --
Saving and
printing
problems
Instrument
not updating
firmware
Countess™
instrument
freezes during
operation
Errors during
updating the
instrument
Incorrect USB
drive
Accidentally
removed the USB
drive
May be using a
corrupted
firmware file or a
damaged USB
drive
Memory is full
Cannot find the
USB drive
Cannot find the
.exe file indicating
that the firmware
is saved
incorrectly on the
USB drive
More than one
firmware version
on the USB drive
See page 24 for a description of error
codes.
Use the USB drive supplied with the
counter or an USB 2.0 drive as some
types of USB drive are not detected or
recorded by the counter.
Do not save too many files in a USB
drive as the counter may slow down
to read the USB drive.
Do not remove the USB drive or turn
off the counter when updating.
Do not remove a USB drive when
saving or reading data as it may
damage the counter.
Download a new version of the firmware
from the website on a different USB drive
and try updating the firmware on the
Countess
Support if the problem persists.
Erase the .CSV file. After performing a
count, go directly to the save menu, press
Start new .CSV file when prompted press
OK to start new .CSV file.
Use the Countess™ USB drive supplied
with the instrument or a USB drive that
you routinely use with the instrument.
Ensure the firmware files (a total of 5 files)
are unzipped on the USB drive as the
instrument cannot access files required for
updating if they are in a folder.
To unzip the files, double click the
icon>click Extract.
Erase all previous firmware versions on
the USB drive and then save the new
firmware version on the USB drive.
™
instrument. Contact Technical
23
Countess™ Automated Cell Counter Error
Codes
Introduction
Position Reason Message Error code
This section describes the error codes displayed by the
Countess
Contact Technical Support (page 26) for details on error codes
and if you need
Camera driver loading fail 0101
™
automated cell counter when it encounters a problem.
to send the instrument back for servicing.
Driver
Preview Camera preview fail 0201
Memory
Camera driver open fail 0102
Camera driver initializing fail 0103
GPIO driver loading fail 0104
Image buffer memory allocation
fail
Camera memory allocation fail
System
Error
(E0101)
Reboot the
device or
refer to
service
personnel.
0301
0302
24
Appendix
Accessory Products
Additional
Products
The following products can be used with the Countess™
automated cell counter and are available separately from
Invitrogen.
For more information, visit www.invitrogen.com
Technical Support (page 26).
Product Quantity Catalog no.
Countess™ Cell Counting
Chamber Slides, 50 Slides
(100 counts)
Countess™ Cell Counting
Chamber Slides, 500 Slides
(1000 Counts)
Countess™ Cell Counting
Chamber Slides, 1250 Slides
(2500 Counts)
Countess™ Cell Counting
Chamber Slides, 2500 Slides
(5000 Counts)
Countess™ Cell Counting
Chamber Slides, 5000 Slides
(10,000 Counts)
Countess™ power cord with
four adapter cords
Countess™ USB drive 1 each C10286
Trypan blue stain (0.4 %) 2 × 1 mL T10282
Countess™ test beads 1 mL C10284
or contact
1 box C10228
10 boxes C10312
25 boxes C10313
50 boxes C10314
100 boxes C10315
1 each C10285
25
Technical Support
Web
Resources
Contact Us
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E-mail:
tech_support@invitrogen.com
Visit the Invitrogen Web site at www.invitrogen.com for:
Technical resources, including manuals, vector maps and
sequences, application notes, MSDSs, FAQs, formulations,
citations, handbooks, etc.
Complete technical support contact information
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dg. 6F
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siness Park
26
Purchaser Notification
Limited Use
Label License
No. 5:
Invitrogen
Technology
The purchase of this product conveys to the buyer the nontransferable right to use the purchased amount of the product
and components of the product in research conducted by the
buyer (whether the buyer is an academic or for-profit entity).
The buyer cannot sell or otherwise transfer (a) this product (b) its
components or (c) materials made using this product or its
components to a third party or otherwise use this product or its
components or materials made using this product or its
components for Commercial Purposes. The buyer may transfer
information or materials made through the use of this product to
a scientific collaborator, provided that such transfer is not for
any Commercial Purpose, and that such collaborator agrees in
writing (a) not to transfer such materials to any third party, and
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Purposes means any activity by a party for consideration and
may include, but is not limited to: (1) use of the product or its
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the product or its components for therapeutic, diagnostic or
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Technologies Corporation will not assert a claim against the
buyer of infringement of patents owned or controlled by Life
Technologies Corporation which cover this product based upon
the manufacture, use or sale of a therapeutic, clinical diagnostic,
vaccine or prophylactic product developed in research by the
buyer in which this product or its components was employed,
provided that neither this product nor any of its components
was used in the manufacture of such product. If the purchaser is
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than for research use please contact Out Licensing, Life
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outlicensing@lifetech.com
Continued on next page
27
Purchaser Notification, Continued
Instrument
Warranty
Invitrogen (a part of Life Technologies Corporation) warrants to the
original purchaser (“Purchaser”) that the Instrument (“Instrument”)
will be free from defects in materials and workmanship for a period
of one (1) year from the date of delivery. Invitrogen agrees, as its sole
responsibility under this limited warranty, and upon prompt notice
of a defect, to repair, replace or credit the purchase price, at its
discretion, any Instrument discovered to be defective within the
warranty period.
Invitrogen will credit the account within 30 days of purchase, if the
instrument is in original condition. After 30 days, Invitrogen will
only replace the instrument for up to a year and no credit will be
issued. Invitrogen will not accept any returned instrument that was
used in HIV or other infectious disease labs.
This warranty does not include repair, replacement, or refund
necessitated by accident, abuse, neglect, misuse, unauthorized repair,
or modification of the Instrument. The warranty will be voided if the
instrument is disassembled or a customer attempted to repair the
instrument. In the event that Invitrogen determines that the
Instrument is in need of repair and not replacement, this Standard
Warranty includes replacement parts and labor for the Instrument.
This Standard Warranty does not include shipment of the Instrument
to and from service location or travel cost of service engineer, the
costs of which shall be borne by the Purchaser.
This Warranty and the remedies set forth herein are exclusive and
in lieu of all other express or implied warranties (including
implied warranties of merchantability, fitness for a particular
purpose and non-infringement), and no other warranties shall be
binding upon Invitrogen. In no event shall Invitrogen be liable for
any special, incidental or consequential damages resulting from
the use or malfunction of this Instrument or the system With
which it is used, even if such damages could be anticipated by Life
Technologies Corporation.
To obtain service during the warranty period, contact Invitrogen
Technical Support for further instruction.
OUT OF WARRANTY SERVICE
Contact Invitrogen Technical Support. We will be happy to assist you
by phone at no charge. Repair service, if needed, will be billed
depending on the parts replaced and labor hours needed to repair
your instrument. You will be billed for shipment of the instrument to
the recommended service facility.
28
©2009 Life Technologies Corporation. All rights reserved.
For research use only. Not intended for any animal or human
therapeutic or diagnostic use.
Corporate Headquarters
5791 Van Allen Way
arlsbad, CA 92008
C
T: 1 760 603 7200
F: 1 760 602 6500
E: tech_support@invitrogen.com
For country-specic contact information, visit our web site at www.invitrogen.com
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