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Author Topic: Carbodiimide Fixation  (Read 4709 times)

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Offline rtcarpenter

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Carbodiimide Fixation
« on: August 27, 2008, 12:57:45 PM »
i have a synthetic molecule which is internalized by receptor mediated endocytosis. the problem is that it's not fixable by aldehydes (it doesn't have any free amines) and tends to diffuse after fixation/permeabilization. It does have some carboxyl groups on it, so i was thinking about trying a carbodiimide fixation for it. Does anyone have any advice/experience with carbodiimide fixation of cell suspensions? thanks!

Carbodiimide Fixation
« on: August 27, 2008, 12:57:45 PM »

Offline CanuckPhD

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Re: Carbodiimide Fixation
« Reply #1 on: August 28, 2008, 02:01:40 AM »
I'm not very familiar with Carbodiimide fixation (In fact I had to look it up to remind myself how it worked). However, there are several other fixation methods to try such as acetone, ethanol and methanol. All of these works as simple coagulation fixatives, therefore lack amine group is not a worry.

How are you growing the cells? If you use any of these fixatives in tissue culture dishes make sure that the dishes are resistant to them. I have been caught adding acetone to TC dishes and watching the plate and my cells dissolve into a mush.


Offline rtcarpenter

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Re: Carbodiimide Fixation
« Reply #2 on: August 28, 2008, 10:11:55 AM »
yeah, organics and plastics are not so much good friends...

i was worried about using fixation methods that would precipitate and dehydrate since I'm also staining for endocytic/lysosomal compartments (currently using LAMP-2 marker). I'd like to be able to see some subcellular morphology...

Re: Carbodiimide Fixation
« Reply #2 on: August 28, 2008, 10:11:55 AM »