Record of cell counts Please note that these counts have all been done with Tryp
ID: 197780 • Letter: R
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Record of cell counts Please note that these counts have all been done with Trypan Blue. Therefore, all cultures have been diluted two-fold (equal volume of cell suspension equal volume of Trypan Blue). Take this into Day Total cytometerVolumeDilution PBS Live Cells/ml Viability (live cells only) Fuchs Fuchs Fuchs Fuchs Fuchs Fuchs Fuchs Fuchs Fuchs Fuchs lml 1ml 1ml lml lml lml lml lml lml 1ml NA NA NA 95 5*1044 7.7*1044 3*1045 #1015 8.3*105 1.9*10*6 6.59*10 6 2.46*10 6 1.95*10 6 25 89 4 132 4 1.5 415 643 659 410 389 16 19 10 98 24 95 15 86 account when determining cell number. Any dilution listed in table is an additional dilution. Live and dead cells listed below represent the total number of cells in 5 large (1 mm) counting squares using a Fuchs-Rosenthal hemocytometerExplanation / Answer
Day
Hemocytometer
Total volume
PBS
Dilution Factor
Live
Average number of cells in one square= Live count/5
Dead
Total cell count= Live + dead
% Viability
Cells/ml Viable
1
Fuchs
1 ml
NA
1
NA
NA
NA
NA
95
1* 105
2
Fuchs
1 ml
0
1
25
5
3
28
25/28*100=
89.28
5*2*1*10000=
1* 105
3
Fuchs
1 ml
0
1
77
15.4
9
86
77/86 *100=
89.53
15.4*2*1*10000=
3.08* 105
4
Fuchs
1 ml
0
1
132
26.4
4
136
132/136*100=97
26.4*2*1*10000 =
5.28*105
5
Fuchs
1 ml
1.5
2.5
222
44.4
9
231
222/231*100=
96.10
44.4*2*2.5*10000
= 2.22*106
7
Fuchs
1 ml
2
3
415
103.75
16
431
415/431*100=
96.29
103.75*2*3*10000
=
6.22* 106
9
Fuchs
1 ml
3
4
643
128.6
19
662
643/662*100=
97.13
128.6*2*4*10000=
1.028* 107
11
Fuchs
1 ml
10
11
659
131.8
14
673
659/673*100=
97.91
131.8*2*11*10000=
2.899 * 107
13
Fuchs
1 ml
6
7
410
82
24
492
410/492*100=
83.33
82*2*7*10000
=
1.148* 107
15
Fuchs
1 ml
5
6
389
77.8
86
475
389/475*100=
81.9
77.8*2*6*10000=
9.33* 106
Dilution factor
Dilution 1: 2= 1ml Sample+1ml PBS.
Dilution factor is 2.
Applying same principle to all dilution factor calculations.
% Viability= Live cell count/Total cell count * 100
Total number of Viable cells= Total number of viable cells per square * Dilution factor *2*10000 cells/ml
You need to multiply both by 2 dilution factor (as it was a two fold dilution) and the additional dilution factor.
Day
Hemocytometer
Total volume
PBS
Dilution Factor
Live
Average number of cells in one square= Live count/5
Dead
Total cell count= Live + dead
% Viability
Cells/ml Viable
1
Fuchs
1 ml
NA
1
NA
NA
NA
NA
95
1* 105
2
Fuchs
1 ml
0
1
25
5
3
28
25/28*100=
89.28
5*2*1*10000=
1* 105
3
Fuchs
1 ml
0
1
77
15.4
9
86
77/86 *100=
89.53
15.4*2*1*10000=
3.08* 105
4
Fuchs
1 ml
0
1
132
26.4
4
136
132/136*100=97
26.4*2*1*10000 =
5.28*105
5
Fuchs
1 ml
1.5
2.5
222
44.4
9
231
222/231*100=
96.10
44.4*2*2.5*10000
= 2.22*106
7
Fuchs
1 ml
2
3
415
103.75
16
431
415/431*100=
96.29
103.75*2*3*10000
=
6.22* 106
9
Fuchs
1 ml
3
4
643
128.6
19
662
643/662*100=
97.13
128.6*2*4*10000=
1.028* 107
11
Fuchs
1 ml
10
11
659
131.8
14
673
659/673*100=
97.91
131.8*2*11*10000=
2.899 * 107
13
Fuchs
1 ml
6
7
410
82
24
492
410/492*100=
83.33
82*2*7*10000
=
1.148* 107
15
Fuchs
1 ml
5
6
389
77.8
86
475
389/475*100=
81.9
77.8*2*6*10000=
9.33* 106
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