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Offline MARS

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BEST ATTACHMENT
« on: April 16, 2008, 12:09:11 PM »
HI EVERYONE.
I WOULD LIKE TO KNOW THE BEST WAY TO AVOID THE CELL LOSS WHEN PERFORMING AN IMMUNOCYTOCHEMISTRY.

I have some experience with immunohistochemistry and now I have to start with cells. I have heard the glass coverslips work much better than the plastic chambers, but I still observe a lot of cells peel off from the glass.
any sugestions in the treatment of the cultures and/or materials would be appreciated
regards.

BEST ATTACHMENT
« on: April 16, 2008, 12:09:11 PM »

Offline CanuckPhD

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Re: BEST ATTACHMENT
« Reply #1 on: April 17, 2008, 07:38:57 AM »
What kind of cells are you staining? Is it an adherent cell line? Are you fixing the cells prior to staining?

If you are trying to stain a cells that are only weakly adherent you will lose many of them and may have to perform cytospins to get them to stick to your slide. Of course if you are staining an adherent cell type (example primary glia cells) you can perform the staining on a glass coverslip.

Good luck.

Offline MARS

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Re: BEST ATTACHMENT
« Reply #2 on: April 28, 2008, 04:35:36 AM »
Actually, I am staining mice embrionic stem cells which are adherent but many of the cells peel off form the glass coverslip during the washes and I would like to try those cytospins you mentioned.
Do you have experience on these cells?
thanks for answering.

Offline ImmunoNYC

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Re: BEST ATTACHMENT
« Reply #3 on: January 19, 2009, 04:11:43 PM »
Glass is generally to be avoided for lightly adherant cells. People in my lab use plastic coverslips (- think Thermanox?) For growing their ECs. Also modifying your staining protocol to make gentler might help. If you want to use coated glass be aware that the coating could affect your ES. However people use gelatin, laminin, fibronectin or even Cell tak.

Re: BEST ATTACHMENT
« Reply #3 on: January 19, 2009, 04:11:43 PM »