JRD409 cell adhesion experiment

Cell adhesion experimental experimental principle Cell adhesion experiments are usually divided into two categories, namely, cells and cell adhesion and cells adhere to matrix. Many cells in the body, such as epithelial cells are fixed to somewhere; other cells, such as white blood cells, active motion, need to continuously regulate cell adhesion. Cell adhesion changes also play an important role in tumor metastasis. The malignant tumor has the ability to isolate from primary tumor and proliferation in vivo, suggesting that these cells have changed mutual identification and adhesion mechanism. Experiments for determining cell adhesion: Mechanical determination of cell adhesion, emission nucleating nucleutin method, cell adhesion, cell separation experiment, and tumor cell aggregate formation experiment.
Experimental step
reagent: FN fibronectin, BSA reagent, 96-well plate, MTT or CCK-8 reagent
1. Pred-taking 96-well plate with 10 u0026 micro; g / ml , 70 u0026 micro; l / well, after 4 ° C, PBS washed 3 times, and then 1% BSA was closed at 37 ° C 1 h, PBS wash 3 times
2. The cells to be tested into the logarithmic growth During the period, digestion cells, suspension cells with serum-free medium, count the blood cell count plate, adjusted in 96-well plates of the pre-patter of 5 * 105 / mL, 5000 cells per well, each set of 3 Repair. 37 ° C Incubate 1 H, PBS wash the unneained cell.

Method 1: 3.5% glutaraldehyde in 4 ° C fixed 0.5 h, PBS wash 3 times; 0.1% crystalline purple color, room temperature stationary 0.5 h, PBS Wash 3 all over, 100 u0026 mu; l 10% acetic acid, 5 ~ 10 MIN, use the enzyme gauge instrument to detect 595 NM absorbance value to indicate how much adhesive cells.
Method 2: Add 100 u0026 Micro; L methanol in accordance with each well, fix 15 min each hole to 100 u0026 micro; l Jimsa staining, dyeing 15 min, PBS washing dry liquid inverted microscope randomly 5 Quantity of the number of skeletons of adhesive cells and statistical results.
Method 3: MTT method or CCK-8 detects the amount of cells.

Customers need to provide
1 customers provide experimental grouping situations and experimental background information, there are specific requirements, please explain in advance;
2 Cell strains (also provided by us), intervention factor raw materials; 3 intervention drugs and concentrations.


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