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Record of cell counts Please note that these counts have all been done with Tryp

ID: 197780 • Letter: R

Question

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 hemocytometer

Explanation / 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