latest video : H&E Staining | Purpose of staining in histopathology | Human Touch in H&E Staining : ru-vid.com/video/%D0%B2%D0%B8%D0%B4%D0%B5%D0%BE-WBUriohMkSM.html
In my lab, we thaw the cryovial, then centrifuge it and remove the media using pipette. After that we add fresh medium in the vial resuspend with the cell pillets.Then transfer it to the 7 ml of media.Directly adding cells with dmso in our lab make our cells dead.Our cel line is HELA adn MDA-231.
This technique is widely accepted when it comes to thawing immortalized cell lines. However, when working with primary cells and stem cells, this procedure is not advised as the cells are very fragile and would benefit from not putting them through excessive stress.
The video is 13 years old, protocol has updated since then. Those days this was the method. Today the protocol is modified so follow the latest procedures. Overall, the basic principle will always remain same. ✌
Thank you for your informative video. I was wondering if 1mM PMSF is added to 10 ml of RIPA lysis buffer. It would be great if you could clarify the exact amount of RIPA lysis buffer.
Reading the protocol - the volume of Trizol needs to be titrated according to the volume of cells to be obtained, and whether the cells have been grown in monolayer or suspension. Trizol is quite powerful so can degrade the cells and RNA altogether. Don't use 1 mL Trizol uniformly - only the volume you need. The volume of Trizol affects the volume of Chloroform, Isopropanol and Ethanol to be added later in the reaction.
Hi, thanks for the video. Please, I have a question: I will like to ask a question about cell treatment experiment with a test drug for 72 h. In such an experiment, do I have to add my drug once and incubate for 72h then check the cell viability or I have to remove the drug solution and replace with the same drug and concentration after each 24h to avoid wrong readings due to degraded drug in wells or evaporation? Please any other person reading the comments can also reply me if you have the correct answer to my question. Thank you
I appreciate the video warning against late behavioral/social interventions in the "Wait and See" method. By then, it is a game of catch up of addressing side effects while trying to get to the root cause. Giving extra support may provide different mileage, but withholding has much more drastic implications.
Hello, I have two questions 1. After collecting my sample and adding Trizol, can i store it in -20C for a month? 2. Why we must change centrifugation 12000g to 7500g in etoh wash step? Can i perform this step with 12000g?
Does the T25 flask has the cap without holes when it arrived ? If yes could be used for cell culture especially if we loose the cap as you probably did before injcubation?
