How to judge the state of cells in the cell culture flask

 The cell culture flask is a square cell culture vessel generally used for the culture of adherent cells. When culturing cells, we need to grasp the state of cells to determine the effect of cell culture. Cell viability is an important indicator.

When culturing cells in cell culture flasks, there are always some cells in the population that die for various reasons, and the percentage of live cells in the total cells is called cell viability. Usually we will use the following methods to detect:

1. The dyeing exclusion method is also a more commonly used method.

(1) Trypan blue method: dead cells are stained blue, live cells are not stained;

(2) Nigrosine method: dead cells are stained black, live cells are not stained;

(3) Eosin Y method: Under the light microscope, the nucleus is blue-black and the cytoplasm is light red. Apoptotic cells are scattered singly in tissues, with dense and condensed nuclear chromatin, nuclear pyknosis, nuclear fragmentation, and the formation of apoptotic bodies.

 FuDau T75 Cell Culture Flasks

2. Fluorescence exclusion method: cells with strong viability can emit strong yellow-green fluorescence; cells with weak viability also emit weak fluorescence; dead cells have no fluorescence.

3. Color reaction between intracellular enzymes and specific reagents:

(1) MTT colorimetric experiment: succinate dehydrogenase can decompose MTT to produce blue crystalline formazan granules that accumulate in and around cells, and the amount is proportional to the number of cells and to cell viability.

(2) Clone formation test: an effective method to measure the proliferation ability of single cells. Clone formation rate = (number of clones/number of inoculated cells) × 100%

The state of cells in the cell culture flask can be judged by the detection of cell viability. The specific method to choose depends on the actual needs.