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Author Topic: IF staining cells on slide  (Read 10517 times)

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Offline apple

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IF staining cells on slide
« on: November 27, 2005, 12:09:02 PM »
I need to do IF staining for cells on slides. My cells are not adherent cells. They stay in media suspension. I don't have cytospin machine. Could anybody suggest a detail protocol including transfering cells onto slides, fixation cells, IF staining cells etc. If I don't want to stain cells on slides directly, can I store the slides ? at what condition? after fixation or before fixation?

Many thanks

Apple

IF staining cells on slide
« on: November 27, 2005, 12:09:02 PM »

Offline richard03

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IF staining cells on slide
« Reply #1 on: November 28, 2005, 08:12:00 PM »
I would go with staining cells in suspension and mount on slides to avoid cells coming off slides.

Offline apple

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IF staining cells on slide
« Reply #2 on: November 29, 2005, 06:04:54 AM »
Hi Richard,

Do you know how to fix cells in suspension? use 4% PFA or cold icetone or methanol?

Thanks

Apple

Offline richard03

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IF staining cells on slide
« Reply #3 on: November 29, 2005, 07:27:32 AM »
I use 4% PFA, sometimes methanol.

Offline apple

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IF staining cells on slide
« Reply #4 on: November 30, 2005, 10:37:28 AM »
Hi Richard,

Could you please give me a protocl for fixation cells in suspension ? at which conc of cell number? for how long? should I leave for air dry for how long? etc..so many details.

how about acetone? some people use -20C acetone for fixation cells.

For cytospin, if I don't want to stain slides directly, I can store slides at -80C. I think it would be better to fix slides firstly then store them at -80C. And when I stain them, I need to bring them to room temperature and fix quick fix again. Is it right?

Thanks

Apple

Offline richard03

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IF staining cells on slide
« Reply #5 on: December 07, 2005, 07:32:00 AM »
Apple,

Check the following link for detailed methods on how to fix cells. Hope this helps.

Immunofluorescence Labeling of Cells

IF staining cells on slide
« Reply #5 on: December 07, 2005, 07:32:00 AM »