Your colleague in the laboratory was able to collect all his

Your colleague in the laboratory was able to collect all his raw data from a growth curve on the Hela S3 cell line, but had to leave unexpectedly and has asked you to perform cell calculations and prepare a growth curve to show your professor. This what your colleague did: he seeded cells into a 24-well plate (allows for replicate sampling and it is much easier to handle one 24-well plate rather than 24 flasks), and on each of the days indicated in the table on the following page, trypsinized and counted live/dead cells from three individual wells to allow for statistical accuracy. The information in the data table includes: day counted, media volume, dilutions, and number of live and dead cells counted (these numbers represent the average count from the three wells counted each day) with a Neubauer hemocytometer. You need to:

a) Perform all necessary calculations to determine cell number (in this case cells/ml equals total cell number). Show all work.

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 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 Neubauer hemocytometer.

b) Make a Growth curve

Day	Hemocytometer	Total Volume	PBS Dilution	Live	Dead	% Viability	Cells/ml (live cells only)
1	Neubauer	1ml	NA	NA	NA	95	1x10^5
2	Neubauer	1ml	0	25	3
3	Neubauer	1ml	0	64	6
4	Neubauer	1ml	0	120	8
5	Neubauer	1ml	1.5	230	6
7	Neubauer	1ml	2	460	15
9	Neubauer	1ml	3	673	10
11	Neubauer	1ml	10	610	15
13	Neubauer	1ml	6	401	45
15	Neubauer	1ml	5	361	100

Solution

Day Hemocytometer Total Volume PBS Dilution Live Dead % Viability Cells/ml (live cells only) 1 Neubauer 1ml NA NA NA 95 1x10^5 2 Neubauer 1ml 0 25 3 89 5 * 10^5 3 Neubauer 1ml 0 64 6 88 12.8 * 10^5 4 Neubauer 1ml 0 120 8 93 24 * 10^5 5 Neubauer 1ml 1.5 230 6 97 80.5 * 10^5 7 Neubauer 1ml 2 460 15 96 184 * 10^5 9 Neubauer 1ml 3 673 10 98 336.5 * 10^5 11 Neubauer 1ml 10 610 15 97 732 * 10^5 13 Neubauer 1ml 6 401 45 89.9 320.8 * 10^5 15 Neubauer 1ml 5 361 100 78 252. 7* 10^5