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Author Topic: IF staining cells on slides  (Read 10273 times)

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Offline apple

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IF staining cells on slides
« on: November 28, 2005, 05:57:14 PM »
I want to do immunofluorescet stain for cells. I don't have cytospin machine. Supplier suggests me to stain cells in suspension with their PE or FICT or Biotin conjugated ab. Then transfer stained cells onto slides manully. My supervisor suggests me to add a drop of cell solution (5-10ul) onto slide first, leave it air dry and then start IF staining. Which way is the best? Also, do I need to permealize cells before applying primary ab?

Many thanks

Apple

IF staining cells on slides
« on: November 28, 2005, 05:57:14 PM »

Offline richard03

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IF staining cells on slides
« Reply #1 on: November 28, 2005, 08:06:53 PM »
Another alternative is to paraffin embed the cells in "HistoGel"  and treat this as regular tissue and processing for paraffin embedding, sectioning and staining. If you can 5 um sections, there is no need to permealize cells before applying primary ab.

Richard

Offline apple

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IF staining cells on slides
« Reply #2 on: November 29, 2005, 06:10:01 AM »
Hi Richard,

Which supplier has this HistoGel?

Thanks

Apple

Offline richard03

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IF staining cells on slides
« Reply #3 on: November 29, 2005, 07:24:04 AM »

Offline ImmunoNYC

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IF staining cells on slides
« Reply #4 on: November 30, 2005, 11:33:19 PM »
I love Histogel, it rocks. :mrgreen:

Offline apple

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a new problem
« Reply #5 on: December 04, 2005, 05:49:27 PM »
I made some cell slides last week. I simply added 5-10ul cell suspension on to Poly - L - Lysine coated slides. After drying the slides for 24hr at room temperature, I checked these slides under normal light microscope.
 I found apart from cell spots there are some stripe pattern background (some looked like tree/leaves/branch stripe pattern). What's this? where are they come from? I did clean my slides in 100% ethanol before adding cell suspension onto them.
Can anybody help? Have you met this problem before?

Thanks very much

Offline ImmunoNYC

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Re: a new problem
« Reply #6 on: December 04, 2005, 08:58:43 PM »
Are you looking at dried slides in the scope? Assuming yes, it sounds like dried poly-l-lysine to me or perhaps dried media or PBS (or whatever buffer the cells were in before drying?

Offline apple

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IF staining cells on slides
« Reply #7 on: December 06, 2005, 06:31:02 PM »
Hi MaximinaNYC,

Yes. I was looking at dried cell spots on my slides. You right. After staining them, I don't see any of this background under microscope.

Thanks

Offline jonnieplumb

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IF staining cells on slides
« Reply #8 on: December 12, 2005, 08:28:40 AM »
Hi can you tell me a little more about histogel.
Is it an embedding compound, can it be used for biopsies etc.
Cheers
J

Offline richard03

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IF staining cells on slides
« Reply #9 on: December 12, 2005, 05:18:35 PM »
Here you can learn more about the Histogel.  It can be used for small biopsies as well.

http://www.rallansci.com/histology/histology.aspx?id=2

Richard

IF staining cells on slides
« Reply #9 on: December 12, 2005, 05:18:35 PM »