Everything counts
But nothing counts like the
Countess II FL Automated Cell Counter
®
The new Countess II FL Automated Cell Counter is a benchtop cell assay
platform equipped with state-of-the-art optics and image analysis software
for rapid assessment of cells in suspension. With three-channel flexibility—
brightfield and two optional fluorescence channels—researchers can count
cells, monitor fluorescent protein expression, and measure cell viability.
®
•Flexibility—choose the fluorescence optical filters for your application
•Value—reusable counting slide reduces consumable costs
•Precision—autofocus designed to improve accuracy and reduce variability
•Simple—easy-to-use touch-screen interface
“I would describe the new
Countess II FL Automated
Cell Counter as accurate,
consistent, and effortless.”
®
Ravi Pathak, PhD
Baylor College of Medicine
2
Life Technologies | Countess II FL Automated Cell Counter
™
®
Clumpy cells?
Not a problem with the Countess II FL Automated
Cell Counter. Whether using automated or manual cell
counting methods, obtaining accurate cell counts in
samples containing clumpy cells is difficult to achieve.
The advanced counting algorithms of the Countess II FL
Automated Cell Counter can identify clear delineations
of cell boundaries within clumps of cells, thereby giving
precise, accurate cell counts even with clumpy samples
(Figure 1).
®
®
Figure 1. Accurate cell counts—even with clumpy samples—on the
Countess II FL Automated Cell Counter. NIH/3T3 cells were mixed 1:1 in
suspension with a 0.4% solution of Trypan Blue Stain (Cat. No. T10282),
and a 10 μL sample was added to a chamber slide (Cat. No. C10228) and
inserted into a Countess II FL Automated Cell Counter. The segmentation
and counting algorithms of the Countess II FL counter are able to resolve
clumps of cells and score them as live or dead using the trypan blue
exclusion method.
®
®
®
Reusable or disposable slides
The Countess II FL Automated Cell Counter offers
the option of using a reusable slide for counting cells,
thereby significantly reducing long-term consumable
costs. For those who prefer the convenience of
disposable slides, the same slides that are used with
the original Countess Automated Cell Counter can
also be used with the the Countess II FL Automated
Cell Counter (Figure 2).
®
®
®
Precise, accurate counts
The Countess II FL Automated Cell Counter contains
advanced autofocusing and counting algorithms
to quickly and accurately identify and count cells
within a population. Upon insertion of the slide, the
Countess II FL counter will automatically focus
on the cells to avoid user variation associated with
manual focusing mechanisms (Figure 3).
Figure 2. Reusable slide available for the Countess II FL Automated Cell
Counter.
®
3.00E+06
®
®
2.50E+06
2.00E+06
Countess® II FL
1.50E+06
Hemocytometer
1.00E+06
5.00E+05
0.00E+00
NIH/3T3
HeLa
HL-60
Figure 3. Precise, accurate cell counts with the Countess II FL Automated
Cell Counter. NIH/3T3, HeLa, and HL-60 cells were mixed 1:1 in suspension
with a 0.4% solution of Trypan Blue Stain (Cat. No. 10282) and counted
using the trypan blue exclusion method. 10 μL were added to a chamber
slide (Cat. No. C10228) and inserted into a Countess II FL Automated
Cell Counter (blue bar) or hemocytometer and counted manually under a
microscope (purple bar). Each bar is an average of 10 counts, with standard
deviation as shown. The autofocusing and advanced counting algorithms of
the Countess II FL Automated Cell Counter give a more precise, accurate
cell count compared to manual counting.
®
®
®
Life Technologies | Countess II FL Automated Cell Counter
™
®
3
Fluorescence counting that fits your experiments
The Countess II FL Automated Cell Counter uses the same fluorescent light
cubes that power the EVOS fluorescence imaging systems. EVOS light
cubes integrate LEDs and high-performance filters in a novel illumination
system that delivers precise control, minimal maintenance, and exceptional
reliability. With interchangeable light cubes, the Countess II FL Automated
Cell Counter was designed to evolve with your changing laboratory needs. As
your experiments change, the fluorescence capabilities of the Countess II FL
counter can be upgraded with new EVOS light cubes.
®
®
®
®
®
®
Full spectrum of fluorescence detection
With over 15 EVOS light cubes from which to choose,
the Countess II FL Automated Cell Counter is not
limited to detection of GFP- and RFP-expressing cells
(Table 1). Looking to measure the transfection efficiency
of cells transduced with an RFP-expression construct
with a blue nuclear counterstain? No problem. Simply
add the RFP and DAPI light cubes, and you can
effectively count the number of cells expressing the
RFP fusion within the total population and accurately
determine the transfection efficiency (Figures 4 and 5).
®
®
Figure 4. Two-color GFP and RFP fluorescent cell counting. 10 µL of HeLa
cells transduced with CellLight Histone 2B-RFP (C10595) and stained
with NucBlue Live ReadyProbes Reagent were added to a chamber
slide (C10228) and inserted into a Countess II FL Automated Cell Counter
equipped with DAPI (AMEP4650) and RFP (AMEP4652) light cubes. The
segmentation and counting algorithms of the new Countess II FL counter
identifies and counts the cells based on their fluorescence properties.
In this experiment, 86% of the HeLa cells within the population were
transduced with the CellLight Histone 2B-RFP construct.
®
®
®
®
®
®
Figure 5. Two-color GFP and DAPI fluorescent cell counting. 10 µL of HeLa
cells transduced with CellLight Mitochondria-GFP (C10600) and stained
with NucBlue Live ReadyProbes Reagent were added to a chamber
slide (C10228) and inserted into a Countess II FL Automated Cell Counter
equipped with DAPI (AMEP4650) and GFP (AMEP4651) light cubes. The
segmentation and counting algorithms of the new Countess II FL counter
identifies and counts the cells based on their fluorescence properties.
In this experiment, 72% of the HeLa cells within the population were
transduced with the CellLight Mitochondria-GFP construct.
®
®
®
®
®
®
4
Life Technologies | Countess II FL Automated Cell Counter
™
®
EVOS light cubes for the Countess II FL Automated Cell Counter
At the heart of the Countess fluorescence technology lie the proprietary EVOS LED light cubes. Each cube contains
an LED, illuminating optics, and filters. Light cubes are user interchangeable, auto-configured by the Countess
system with plug-and-play capability. The wide variety of light cubes available provides flexibility for multiplefluorescence research applications, like cell viability analysis (Figure 6).
®
®
®
®
®
Table 1. Common fluorescent light cubes available for the Countess II FL Automated Cell Counter
®
Light cube
Excitation (nm)
Emission (nm)
Common compatible dyes/fluorescent proteins
DAPI
357/44
447/60
DAPI, Hoechst, BFP
TagBFP
390/18
447/60
TagBFP
CFP
445/45
510/42
ECFP, Lucifer Yellow
GFP
470/22
510/42
GFP, Alexa Fluor 488, SYBR Green, FITC
YFP
500/24
524/27
EYFP, acridine orange (+DNA)
RFP
531/40
593/40
RFP, Alexa Fluor 546, Alexa Fluor 555, Cy 3, DsRed,
Rhodamine Red, dTomato
Texas Red
585/29
624/40
Texas Red , Alexa Fluor 568, Alexa Fluor 594,
MitoTracker Red, mCherry
Cy 5
®
628/40
692/40
Cy 5, Alexa Fluor 647, Alexa Fluor 660, DRAQ5
Cy 5.5
655/46
794/16
Cy 5.5
®
®
®
®
®
®
®
®
®
®
®
®
®
®
®
®
For a complete list of available common and specialty EVOS light cubes, go to lifetechnologies.com/evoslightcubes
®
Figure 6. Cell viability. Heat-treated HL-60 cells were combined with
non-treated cells at approximately a 15:85 ratio. To assess cell viability
of the population, cells were stained with a combination of NucBlue Live
and NucGreen Dead 488 stains. 10μL of cells were added to a chamber
slide and inserted into a Countess II FL Automated Cell Counter
equipped with DAPI (AMEP4650) and GFP (AMEP4651) light cubes. As
expected, 14% of the HL-60 cell population were dead as indicated by
positive NucGreen Dead 488 (green) staining.
®
®
®
®
Life Technologies | Countess II FL Automated Cell Counter
™
®
5
Table 2. Cell lines validated on the Countess II FL Automated Cell Counter
®
6
Cell type
Animal
Organ
A431
Human
Skin
Adipocytes
Human
Adipose-derived stem cells
Aortic smooth muscle
Human
Smooth muscle
CHO-M1WT2
Chinese hamster
Ovary
CHSE
Chinook salmon
Embryo
COLO-205
Human
Colon
COS-7
African monkey
Kidney
HEK-293
Human
Kidney
HeLa
Human
Cervix
HepG2
Human
Liver
HL-60
Human
Blood
J774A.1
Mouse
Blood
Jurkat
Human
Blood
K562
Human
Bone marrow
MCF7
Human
Breast
MRC-5
Human
Lung
NIH/3T3
Mouse
Embryo
PC-12
Rat
Adrenal gland
Pulmonary artery endothelial cells
Human
Blood vessel
Pulmonary artery smooth muscle
Human
Smooth muscle
SF-21
Insect
Ovary
U266
Human
Blood
U-2 OS
Human
Bone
Umbilical vein endothelial cells
Human
Blood vessel
Life Technologies | Countess II FL Automated Cell Counter
™
®
Table 3. Countess II FL Automated Cell Counter technical specifications
®
Processing time:
<10 seconds
Sample concentration range: 1 × 10 to 1 × 10 cells/mL
4
Technical specifications
7
Particle/cell diameter range: 10–60 μm
Required sample volume:
10 μL
Firmware: Countess Automated Cell Counting Platform
®
USB drive: Software
Optics
Optics:
Camera:
Instrument type:
Instrument dimensions:
Weight:
3 channels (brightfield and 2 slots for EVOS LED light cubes)
®
5 megapixels, 2.5x optical magnification
Benchtop cell counter and suspension cell–based assay platform
9" (W) × 5½" (D) × 9" (H)
228.6 mm (W) X 139.7 mm (D) x 228.6 mm (H)
8 lb
Operating power: 100–240 VAC, 0.58 A max
Physical characteristics
Frequency:
50/60 Hz
Electrical input:
12 VDC, 2 A
Installation site:
Indoor use only, Class A environments (i.e., nonresidential or
light industrial); Pollution degree 2.
Operating temperature: 10–40˚C
Operating humidity:
Material:
Reusable analysis slide
Dimensions:
Chamber volume:
Material:
Disposable analysis slide
<80% (noncondensing)
Glass
Slide: 32 mm (W) × 76 mm (D) × 3.9 mm (H)
Coverslip: 20 mm (W) x 26 mm (L) x 0.5 mm (H)
10 μL
Poly(methyl methacrylate) (PMMA)
Dimensions: Slide: 25 mm (W) × 75 mm (D) × 1.7 mm (H)
Chamber volume:
10 μL
Life Technologies | Countess II FL Automated Cell Counter
™
®
7
Need a simple, brightfield cell counter? Check out the Countess II Automated Cell Counter.
®
Feature
Countess II
Countess II FL
Autofocus
✔
✔
Touch-screen
✔
✔
Brightfield counting
✔
✔
Trypan blue counting
✔
✔
Disposable slide
✔
✔
®
®
Reusable slide
✔
Fluorescence compatible/upgradable
✔
Uses EVOS light cubes
✔
®
Find out more at lifetechnologies.com/countessII
Ordering information
Product
Quantity
Cat. No.
Countess II FL Automated Cell Counter
1
AMQAF1000
Countess II Automated Cell Counter
1
AMQAX1000
Countess Cell Counting Chamber Slides
50
C10228
Countess II FL Reusable Slide
1
A25750
Countess II FL Reusable Slide Holder
1
AMEP4746
EVOS Light Cube, DAPI
1
AMEP4650
EVOS Light Cube, TagBFP
1
AMEP4668
EVOS Light Cube, YFP
1
AMEP4654
EVOS Light Cube, GFP
1
AMEP4651
EVOS Light Cube, RFP
1
AMEP4652
EVOS Light Cube, Texas Red
1
AMEP4655
EVOS Light Cube, Cy 5
1
AMEP4656
EVOS Light Cube, Cy 5.5
1
AMEP4673
®
®
®
®
®
®
®
®
®
®
®
®
®
®
®
®
Find out more at lifetechnologies.com/countessII
For Research Use Only. Not for use in diagnostic procedures. © 2014 Thermo Fisher Scientific Inc. All rights reserved.
All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Cy is a
registered trademark of GE Healthcare. DRAQ5 is a registered trademark of Biostatus Limited. CO012822 1214
