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immunofluorescence


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4 replies to this topic

#1 zienpiggie

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Posted 05 April 2011 - 12:52 PM

Hi all,
I am relatively new to the immunocytochemistry technique and I need to use them for my cells attached on 8 chamber slides. I am thinking of using fluorescence secondary antibody for detection but then realized I will be finished with the experiment on saturday and I don't know if I could really keep them for few days before observing them in the microscope? How stable are these fluorophores? If anyone could englighten me in this area I would be really grateful!

Thank you

#2 bob1

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Posted 05 April 2011 - 04:06 PM

If you keep them in the dark and the samples are mounted (coverd by a coverslip), and the edges sealed, then they should be fine for a couple of weeks or more in the fridge. You can even freeze the slides and use them indefinitely.

The fluorophores do fade quite quickly under normal fluorescent light tubes, and very quickly under the microscope - so be careful with the amount of light you let get onto the slides.

#3 zienpiggie

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Posted 06 April 2011 - 09:39 AM

Thanks so much Bob1!!

I am wondering too, my protocol calls for blocking at 37C for 1h. Do you happen to know if I can block overnight at 4 C instead?

#4 bob1

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Posted 06 April 2011 - 07:10 PM

Should be fine, blocking is one of the few things that rarely(!) goes wrong, no matter how long you do it for.

#5 zienpiggie

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Posted 08 April 2011 - 01:15 PM

Thank you Bob1! I really appreciate it!




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