Primary cytoskeleton staining method
Microwire display method steps:
1. Rinse the primary cultured cells of the coverslips with PBS solution for 3 times each time for 30 s;
2. Immobilize primary cells with 2% formaldehyde/PBS for 3 min;
3. Treated with 0.5% trinitrotoluene/PBS 3 times for 10 min each time;
4. Rinse PBS 3 times;
5. Rhodamine-labeled phalloidin (1:10) was reacted for 15 min at room temperature;
6. Rinse PBS 3 times;
7. 60% glycerin + fluorescent anti-quenching agent seal;
8. Observation by fluorescence microscope;
Micropipe display method:
1. Rinse the primary cells of the coverslip 3 times with PBS solution for 30 s each time;
2. Fix with 3% formaldehyde/PBS for 20 min at room temperature;
3. Rinse again with PBS solution for 3 times, each time for 1 min, and finally blot excess PBS solution with filter paper;
4. Put in cold acetone (-10-20 ° C) for 7min;
5. Rinse 3 times with PBS for 1 min each time, and finally blot the excess PBS solution with filter paper;
6. Drip a primary anti-tubulin antibody at a concentration of 0.1-0.2 mg/ml into the center of a clean disc with a diameter of 6 cm, and let the small cover cells face down on the antibody solution and cover the plate at 37 ° C. Placed in the incubator for 45 min - 1 h;
7. Slowly add PBS to the dish at a constant rate, so that the cover sheet floats on the liquid surface, pinch it out with tweezers, and process according to step 3;
8. Drop a drop of fluorescently labeled secondary antibody at a concentration of 0.1-0.2 mg/ml into the center of the clean dish, and thereafter proceed as in step 6;
9. Follow steps 7 and 3;
10. Drop 90% glycerol/PBS at pH 8.5 on a glass slide and cover the primary cover of the small cover sheet on the large cover slip;
11. The cover sheet is placed under a fluorescence microscope;
Royal Jelly is also called Bee Milk. The fresh royal jelly is slightly ropy milk paste substance; it is the excretive mixture of nutrition gland and maxilla gland of the head of little worker bee. The worker bees use this to feed the 1-3 days` worker bee larva and drone larva, 1-5.5 days` queen bee larva and queen bee in the oviposition period. The royal jelly is the biologic product containing very complicated active elements which contains almost all the nutrition elements required by the growth of the human body.
Royal Jelly
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