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Countess Cell Counter FASEB 2009
Countess Cell Counter FASEB 2009
Abstract Figure 1 – Counting variation Figure 2 – Comparison of inter- and intra- Figure 3 – Automated cell counting viability determination Results
between traditional glass and user error for both manual and across multiple cell types
The hemocytometer is the most widely used The experiments shown were used to compare
disposable plastic hemocytometers automated cell counting methods the performance of manual cell counting using a
device for determining cell concentrations,
requiring consistent criteria and tenacity to
Viability Comparison hemocytometer to the CountessTM Automated
Hemocytometer Counting Comparison
obtain measurements correctly and Inter- User Comparison Cell Counter.
100
Primary Cells:
reproducibly. As an alternative, the Countess™ 1.40E+06 Bead Standard
1.40E+06 Adipocyte •Accuracy of the instruments was determined by
Automated Cell Counter employs digital
range of cell concentrations. Using a Student’s bead sample by three different, trained users. Each •Inter-user variation of total cell count was
slip at the correct depth. Three preparations and This experiment was used to judge the accuracy of the CountessTM instrument
t-test, the results showed the CountessTM chamber was counted by each of the users. The same shown to be less than 1% using the CountessTM
counts were performed for each type using a viability readings. Ten cell lines were counted on the Countess™ Automated Cell
instrument measured cell concentrations and work flow was followed for the hemocytometer counts. instrument and as much as 5.5% for trained
standard latex bead sample (Coulter bead standard, Counter – 4 primary lines (keratinocyte, HPAEC, HASMC, and adipocyte), 5 adherent
viability as accurately and precisely as the The error bars show differences in user count results, users counting manually.
9.57x105±10%) following the manufacturer’s cell lines (C2C12, A431, MRC-5, MMM, and COLO-205), and 1 cultured suspension
disposable hemocytometer. Additionally, the suggested protocol. The results show a difference of while the column heights show differences in sample line (K562). For each cell line five samples were prepared using different ratios of Conclusions
effective concentration range for the 30% and 5% from the expected value for the glass preparation for each instrument. As shown in the chart live cells and heat killed cells, to represent theoretical viability points ranging from
CountessTM instrument was two times greater and disposable hemocytometers, respectively. Error above, the variation in total bead counts between users is 0% to 100% live cells. One chamber slide was prepared for each sample and
than the hemocytometer, and the viability bars show one standard deviation above and below as much as 5.5% when using the hemocytometer, and Due to the critical nature of accurate cell
counted three times each using the CountessTM instrument. Counts of ten squares
range matched the hemocytometer. The the mean concentration for each hemocytometer less than 1% when using the Countess™ Automated Cell counts in biochemical and cellular assays, it is
using disposable hemocytometers were performed for each sample.
CountessTM Automated Cell Counter overall type. These data, combined with ease of use Counter. Sample preparation difference was approximately important that the cell counts be easy to
produces results much more rapidly without comparisons, resulted in the use of only the 12% for the CountessTM instrument, and approximately perform, be accurate, and give reproducible
the problem of operator tedium and fatigue or disposable hemocytometer in subsequent 15% for each of the hemocytometers. results. We have shown that when compared
compromised accuracy and precision. hemocytometer comparison experiments. to manual counting using a disposable
hemocytometer, the CountessTM instrument
Figure 4- Accuracy and Precision- Automated cell counting Figure 5-Effective concentration range-Automated cell counting compared to a gives more precise and as accurate results. The
compared to a traditional hemocytometer traditional hemocytometer concentration range of the CountessTM
instrument extends much higher than what a
Counting Comparison- Keratinocytes Viability Comparison- Keratinocytes A serial dilution series of SF-9 cells hemocytometer user could count by eye. This
A B Dilution Series: SF-9 Cells
was made starting with a fresh removes the need for dilutions prior to
Countess
1.20E+06 Countess 99.00
Instrument 4.00E+06 Countess 1 highly concentrated cell sample. counting, thereby improving work flow. Cell
Instrument
1.00E+06 Hemocytometer 97.00 Hemocytometer Countess 2 Three CountessTM slide chambers viability readings were shown to be equivalent
Concentration (cells/mL)
95.00 Measured Concentration (cells/mL) 3.50E+06 Countess 3 were counted three times each for to those determined visually and quantified
8.00E+05 Hemocytometer 1 all of the samples in the dilution using a hemocytometer. Inter-user variation is
% Viability
3.00E+06
6.00E+05
93.00
Hemocytometer 2 series. A manual count of ten also markedly decreased by using the
91.00
2.50E+06
Hemocytometer 3 squares was performed using a CountessTM instrument. These experiments
4.00E+05
89.00 disposable hemocytometer three were performed using over 25 different cell
2.00E+05 87.00
2.00E+06
1.20E+06
times for each sample below 1x106 types, including primary cell lines, with similar
1.00E+06 cells/mL. Due to the density of the results. The Countess™ automated cell counter
1.50E+06
0.00E+00 85.00 8.00E+05 cells, it was difficult to manually streamlines the enumeration of cells to obtain
Instrument Instrument
1.00E+06
6.00E+05
count the samples using the quicker and more reliable results which reduces
The Countess™ Automated Cell Counter hemocytometer for cell
Three separate Countess™ counting chambers were loaded with primary keratinocyte user fatigue and tedium. As is evident in these
4.00E+05
uses the trypan blue dye exclusion method concentrations above 1x106 experiments, automated cell counting improves
2.00E+05
cell samples and counted three times each for a total of nine measurements. These 5.00E+05
to calculate cell concentration and provide 0.00E+00