How to Freeze Cell Cultures

Today I'm gonna show you guys how to freeze yo cells maybe you don't need them right now, maybe you need them in the future, but in any case freezing your cells is a good idea because you have a bank to work from so my man Sam here is gonna show you what he's working on Alright so we're gonna be getting our freezing media its 10 percent DMSO 90 percent DMEM we get this, we hand it to Negin so she can sterilize it. You get your cells from the incubator its at 5% CO2, 37 degrees (Celsius) come check it examine the general cell morphology and confluency to determine whether the cell culture is suitable for freezing okay event redundant ship artist at ref held so lately uh... fantastic self c hoti it's good ended up here look great well kit using a sterile technique to media experience the media from the t twenty five points and an air friendships it one-and-a-half ml pitfall kit and that they are abdicated angry eight itself act that were there relief entrance exam below the trips into certain cells for approximately twenty seconds this will allow the self to start detaching from circus what do not work shirts and efficient occur to belong as this may inadvertently killed the cells strips in his attacks into the cells just a moment you can now see that the cells were much more rounded faulty you know that the cells should still be a cheery to the flats yourself it's attached centrifuges themselves into a pilot and the suspension of the sample will be required as removal of chips india aspiration is required as part of the freezing method okay uh... fix it and that freeze this t trainee packing took quite a while so i'm going to head want to know up freezing media so antoinette right what's polled surface at the plate to make sure it did all the stuff came out sarah paralympic games for us and viral we for about just itself an if so sent back a second opinion that i think we were at the restraint both program which is these are c the cell and very hasset snowman event it affect their innocent presley for play for us and that say state itself slowly mister frosting isn't up all one hundred percent super bowl it's slowly piece about some inside at a rate of about decrease at sears per minute lifestyles they should be god very rapidly to prevent excusal from h and i could potentially damaged yourselves thank you himself like that make things better slide registered it and and then take that to make the leaders of the treadmill trypan blue sits at the disease to see in the living cells this is compared against the debt cells and the cell cancer is able to give you arrived estimate of how many cells or present per milliliter in your samples inject n lekker leaders of the mixture into the cell culture cartridge take the cartridge and then inserted into the southwest machine golden start processing this all accounts she will then give you read out of how many cells are pretty mail leader in do sample for cleanup issue you remove any equipment emedia or other tools email used there's any media or other materials that require refrigeration back of their future dispose of any disposables inappropriate receptacles set up any ballots and becky answer pumps that may have been turned on during your work and finally sprayed on your work area with ethanol univ x's uphold wipes was the bsc churn up the lights and turn on the you'd be like what you guys i hope widespread aren't hammy happier increment old uh... he this