Measurement of cell division index
Measurement of cell division index
1. Principle
The ability of cultured cells to grow, divide and multiply in vitro can be expressed by the division index. It has a certain relationship with the growth curve. For example, as the division index continues to increase, the cell enters the exponential growth phase.
The division index refers to the percentage of dividing cells in the cell population. It is an important indicator for determining the cell cycle and an important basis for selecting cells in different experimental studies.
2. Instruments, supplies and reagents
1. Instruments and supplies: CO2 incubator, ordinary microscope, cell culture blood, coverslip, pipette
2. Reagents: culture medium, pancreatin, methanol, glacial acetic acid, Giemsa stain
3. Operation steps
1. Digest the cells and connect the cell suspension to a Petri dish containing a coverslip.
2. Incubate in a CO2 incubator for 48 hours to grow cells on the coverslip.
3. Take out the cover sheet and operate in the following order:
Rinse with PBS for 3 minutes → methanol: glacial acetic acid = 3: 1 fixation solution for 30 minutes → Giemsa solution staining for 10 minutes → rinse with tap water.
4. After the coverslips are dried, they will be snapped back on the slides for microscopic examination.
5. Calculation:
Division index = number of dividing cells / total cells × 100%
4. Precautions Actions should be taken lightly to avoid the cells on the cover slip.
Attachment: Giemsa dye preparation:
Weigh 0.5 g of Giemsa powder, grind with a few drops of glycerin, and then add glycerin (to make the total amount of added glycerin 33ml). Incubate at 56 ° C for 90-120 minutes. Add 33ml of methanol and store in a brown bottle. This is Giemsa stock solution.
Dilute with PBS as required. Generally diluted 10 times.
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