Determination of cell growth curve
[Determination of Cell Growth Curves] Master the principles and methods of drawing cell growth curves.
Strusion of Cell Growth Curve]
The growth curve is a common method of determining the number of cellular growth, and is an important indicator to determine cell viability. It is the basics of cultured cell biological characteristics. One of the parameters. A after the cell passage, after a short suspension, the wall is subjected to a long short, and the long-lasting exponential growth period is entered. After the cells reaches the saturation density, the growth is stopped, enters the platform, and then degenerates. In order to accurately describe the dynamic changes in the number of cells during the entire process, the cells are required to count the cells. Typically count for 6 days. Generally, 3 parallel repetitions are set up each experiment.
1, Experimental Material:
HELA Cell line.
2, experimental reagents:
DMEM medium (penicillin, streptomycin, 10% serum), 0.25% trypsin solution, cener blue staining solution.
3, Experimental Instrument:
Ultra-clean workbench, CO2 incubator, inverted microscope, microtubric, blood cell count plate, culture bottle, high spirits, pipette, waste Cylinders, cover slides, 24-well plates.
1, a cellulated cell, and counts the cell suspension with medium. According to the result of the cell count, in accordance with 5 × 104 ml of cultured in a 24-well plate, 21-well cells were co-inoculated. 1 ml hole. The remaining three holes can be used in the biochemical control.
After 2,24h, the number of cells was recorded daily, and 3-well cells were taken each time, the count was performed. Calculate the average. Continuous count 7D. The cells were added to a trypsin to add a quantity medium, mixing with a cell suspension. Take a small amount of suspension with the ceramic phenol blue 11 mix. Take a clean blood cell count plate, cover the cover sheet, and add the cell suspension from both sides of the cover. Between the count plates and the cover sheet. Note that the drip is not bubbles, and it is not too much, otherwise the cover is floated and the cell count is not allowed.
3, under low mirror, living cells are not colored, and the colored cells colored by the platform are dark blue, and it is unhealthy or dying cells, and cannot count. Find the square on the count plate, calculate the total number of total cells in the quarterg, and the larger sides are calculated only, and the cell density calculation formula in the upper and lower cell suspensions is: Cell number 1 (L u003d (4) (4) X104X2
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