How to Freeze Cell Cultures


Uploaded by davidbanes on 29.11.2012

Transcript:
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
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using a sterile technique to media experience the media from the t twenty
five points
and an air
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one-and-a-half ml
pitfall
kit
and that they are abdicated angry eight itself
act that were there
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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
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that i think we were at the restraint both
program which is these are c
the cell
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slowly
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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...
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