Thank you! Hamocytometer grid # of cells/mil+ (Average) (DF) (10\") If multiple

Question

Thank you! Hamocytometer grid # of cells/mil+ (Average) (DF) (10") If multiple DFs, # of cells/ml-average) (DF1 ) (DF2) (DFn) ( 104) # of cells/pl.-(Average) (DF) (10') If multiple DFs # of cells/juL = (Average) (DFI) (DF2) (DFn) (10') (1) To prepare for cell seeding, you washed GBM cells (grown in T-75 flask) with RT 1X PBS and trypsinized the cells with 1mL of 0.25% Trypsin. EDTA for 5 min @ 37°C. You then neutralized trypsin solution with 5mL of cell culture media. You transferred the 5mL cells in suspension to a 15mL conical tube, transferred 5OuLto a 1.5ml microcentniuge tube, and mixed with 50? L trypan blue. You then transferred 10?L of the cell:trypan blue mixture to the hemocytometer and counted 750 cells from the four gray corner squares. Average # of cells DF . # ofcells per ?L . # ofcells per mL = The total # ofcells obtained from the T-75 flask so you took 10?L from the initial cell:trypan blue mixture and mixed with 100?L of trypan blue, counted again and obtained a total of 250 cells from the four gray corner squares on the hemocytometer. What would be the # of cells per ?L? (2) Then it occurred to you the cell stock is too concentrated which may affect count accuracy,

Explanation / Answer

Answer:

1) Based on the given information:

2) Based on the revised dilution scheme:

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