Question

1. You have a T25 flask that is 90% confluent. You are asked to freeze all of the cells in your flask at concentration of 1 x 106 cells/ml of freezing mix (using 1 ml cell suspension per freezing cryovial). In order to harvest and count your cells, after PBS rinse, trypsinize the cells with 1 ml 0.5% trypsin/EDTA, when cells detached, quench the trypsin with 4 mls of media, mix well. Take a 0.1 ml sample of the cells to mix with 9.9 ml PBS, count using coulter counter. If sample read 3 times using coulter counter and reads were 5600, 6000, 5800 cells/ml; A. What is the average cell concentration for the sample? (1 point) B. What is the total number of cells harvested from the flask? (1 point) C. What chemical in freezing media prevents ice crystal formation? (1 point) D. How many ml of freezing media will you use to freeze all your cells? And, how many cryovials and what volume per vial? (2 points)
hey PLEASE HELP ME SOLVE FOR ALL PARTS OF THIS QUESTION. im struggling with all parts (a,b,c,&d) please explain all parts of the question. and circle the answer. thank you so much!!

Answer 1

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- The given total number of cells total sumber of calls harrested boud is having 5 so club. the so, 5ond (5000ml) is having 5000x580. -)2.01000,000 cleb. - C/ The given chemical freering Duso is used freering media to prevent the crystal formation, : Honce the cruation u proved Scanned with CamScanner