Abcam Ad

Author Topic: cytospin and cytokine  (Read 6859 times)

0 Members and 1 Guest are viewing this topic.

Offline dato

  • Trainee
  • **
  • Posts: 5
cytospin and cytokine
« on: August 11, 2005, 04:58:43 PM »
Hi

I am doing cytokine (IL-12p40 and IL-12p70) IHC on tissue(spleen) or cytospin (BMDC). for tissue (frozen section), there is usually no problem but cytospin bothers me. I don't have good reprodicible staining. In  general,I really don't know if it is correct to fix the cells first and then cytospin or opposite. I think, the 2nd option is better because if you fix (2% PFA) and then cytospin and air-dry, you don't know how tissue will react to this treatment. I did fixation prior to cytospin primarily to avoid any loss of cytkine during cytospin.But I don't  know if this is really valid assumption that without fixation if you cytospin the cells, you will looose the staining.So who has a broad experience with cytokine staining on cytospin preparation?

david

cytospin and cytokine
« on: August 11, 2005, 04:58:43 PM »

Offline excalibur

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 1109
    • http://www.excaliburpathology.com
cytospin and cytokine
« Reply #1 on: August 11, 2005, 10:38:10 PM »
Spin first,  then fix.

Fresh tissue sticks much better.

We even had our surgeons fooled for a while that a frozen could not be done on fixed tissue  :twisted: Of course it can, you just need to used subbed slides, so the tissue will adhere. Subbed slides-$300/case & regular slides-$100/case.

Anyway, the surgical staff always made sure the specimens needing frozens for diagnosis while the patient was still under anesthesia came to the lab fresh.  :wink:
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Offline braz

  • Trainee
  • **
  • Posts: 8
cytospin and cytokine
« Reply #2 on: April 10, 2006, 11:31:13 PM »
In the case of cultured cells, what would you spin them in, for later fixation and immunocytochemistry? Culture media? PBS? 95% ethanol? Cytospin collection fluid?    :?:

Offline excalibur

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 1109
    • http://www.excaliburpathology.com
cytospin and cytokine
« Reply #3 on: April 11, 2006, 07:43:14 AM »
95% Alcohol will act as a fixative too. Use PBS or the cytospin solution.
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

cytospin and cytokine
« Reply #3 on: April 11, 2006, 07:43:14 AM »