How to Change Cell Culture Media



to change the media in a tea seventy-five flasks begin by placing the appropriate media in the warm water bath to rise the temperature of the media the water bath should be at 37 degrees Celsius this should take about five to ten minutes once the media has warmed up to the appropriate temperature begin by preparing two biosafety cabinets before working in the biosafety cabinet make sure you're wearing a laboratory coat and gloves make sure the blower switch and light switch are both turned on and raise the sash to the marked level line the alarm sounding is normal and should stop after a few seconds if the alarm is still sounding make sure that the size is low the sash level line the biosafety cabinet is an area where perform sterile cell work therefore it is necessary to maintain proper sterile technique spray 70 percent isopropyl alcohol on your gloves and rub them together until dry spray the outside window of the glass and armrests and wipe down the glass and armrests with a cuboid then throw the kimwipe away next continue to spray as a propyl alcohol in the inside of the biosafety cabinet workspace and the inside window this is a standard procedure for any work in the biosafety cabinet and it is necessary to begin all procedures in this way take the media bottle out and spray the entire bottle with isopropyl alcohol and wipe it down with a cuboid immediately after wiping it down place the media bottle under the hood it is important to note that anything that is to be used on cells must be sterile the sterile objects are never to be opened or exposed anywhere outside the biosafety cabinet take out your cheesy 75-plus with cells from the incubator be very careful not to contaminate anything when placing or removing items from the incubator do not cough or sneeze any better to ensure extra caution do not talk in the incubator doors open you will need two ten milliliter individually CIO's pipettes give them a quick spray and wipe them with your hands and place them in the loot begin by twisting the cap of the T 75 claws place the cap face down in the back of the engine where ever there is an open container be careful not to wave your arms or hands over the continue now grab the ten milliliter pipette open it halfway and attached it to the pipette gun adjusting the pipette while it's still in its casing that you can read them the leader markings remove it the wrapping to being extra careful not to touch the tip of the pipette carefully place the pipette into the flask making sure the tip of the pipette does not come in contact with anything else in the hood aspirate all the media using the top button of the pipette gun take the pipette out and discard the media in the appropriate way speaker leaving the bottom button of the pipette gun remove the pipette and place it in the appropriate waste container open the media bottle and attach a 10 millimeter pipette to the pipette gun and draw up ten milligrams of one place the cap back on the media bottle to avoid contamination release the media into the TV 75-plus gently to avoid serving the attached cells inside discard the pipette place the cat back on the glass and tighten up the media bottle cap place the plastic back inside intubated seen up the biosafety cabinet is free the work areas using x cup of alcohol and wiping it down with two light close the stash

21 thoughts on “How to Change Cell Culture Media

  1. You should need to edit the title or add in description section about the cell type whether it is attached or suspension cell line. Because if it was suspension cell line, all the cell would be discarded as well by removing the media at first place.

  2. Why are you wasting your time spraying the pipette packaging? You aren't
    going to get it close to being completely sterile, and that wrapper
    should never be over any of your open containers, anyway.

  3. great video indeed 🙂
    am just start doing my phd work …. and this video will help me 🙂
    sterile work = great data/result 🙂 cheers 🙂

  4. The video is educative and interesting. However, if you are not of maintaining the sterility or not too sure of your working environment please revert to using flame at every point of your work.

  5. what kind of bracelet was he wearing? is it sterile? I think not… for shame, sir. for shame.. 🙁 not 😀

  6. KS LEE you mean open the lid slightly of the T-75, after placing in the incubator for air flow? It may be the T-75 where the cap has a filter, so you do not have to loosen it.
    Cheers

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