Experimental steps:
1. Put the sterile Microscope Slides Laboratory in the cell culture dish and take it out after the cells grow to semi-confluent. You can also choose to dye directly in a petri dish or a culture plate, and there is no need to put in a sterile Slide Glass Microscope at this time;
2. After washing with PBS, fix with cold acetone for 10 minutes, and then wash with PBS 3 times after fixation. You can also choose to fix with 95% ethanol or formaldehyde, and it is best to place it at 4°C during fixation;
3. Prepare the incubation solution:
3% β-Sodium Glycerol: 5ml
2% sodium barbiturate: 5ml
2%CaCl2: 10ml
2%MgSO4: 1ml
Distilled water: 10ml
4. Put it in the incubation solution, incubate at 37°C for 4-6 hours, rinse with PBS 3 times;
5. Soak in 2% cobalt nitrate for 5 minutes and rinse with PBS for 3 times;
6. Soak in 1% ammonium sulfide for 2 minutes, rinse with PBS 3 times, dry naturally, and seal;
7. Observe the staining result under the microscope. The positive reaction in the cytoplasm showed gray-black particles or massive precipitates.
Precautions:
1. Indirect flushing should be done during flushing. If it is a microscope glass slide, pipette in the liquid around the slide. For indirect flushing, the petri dish or culture plate should be slowly piped around the culture dish or culture plate with a pipette tip, so that the cells are It is not easy to take off the film or blow it off.
2. Cobalt nitrate with 6 water, ammonium sulfate with 7 water, do not need to remove the weight of water when weighing, directly weigh. 1% Ammonium sulfide is directly mixed with the stock solution to make a 1% solution, the stock solution is regarded as 100%, and it is not calculated by sulfur.
Post time: Dec-17-2021