Author Topic: paraffin sectioning of mouse eyes  (Read 2814 times)

0 Members and 1 Guest are viewing this topic.

Offline MSturm

  • Newbie
  • *
  • Posts: 2
paraffin sectioning of mouse eyes
« on: July 07, 2011, 12:14:50 PM »
We are sectioining mouse eyes using plus slides.  We are using these for extensive immunohistochemistry procedures but are having difficulty keeping the eyes on the slides for the entire procedure...ususally about right after the primary the eyes start curling and then sliding off the slides.  We bake them extensively before we start our procedure...overnight, and then some for even over the weekend.  Anyone have any suggestions for us?
Thanks
Mary Sturm
University of Iowa
mary-sturm@uiowa.edu

paraffin sectioning of mouse eyes
« on: July 07, 2011, 12:14:50 PM »

Offline gula

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 316
Re: paraffin sectioning of mouse eyes
« Reply #1 on: July 07, 2011, 02:33:34 PM »
Do you dry the slides vertical or horizontal? Water under the slides can be trapped, if you dry them horizontal. This inhibits the binding on the glass surface.

Superfrost plus have an expiration date. Are yours still ok?

gula

Offline DCundiff

  • SuperTech
  • ****
  • Posts: 23
Re: paraffin sectioning of mouse eyes
« Reply #2 on: July 20, 2011, 11:58:56 AM »
I had a similar problem with some of my brain tissue.  Are you "baking" overnight at 37C?  I dry at room temperature for at least 30 minutes (vertically) - using Superfrost positive slides- and then overnight at 37C.  The next day I incubate at 60C for one hour prior to starting the protocol.   

Offline richard03

  • Administrator
  • GoldMember
  • *****
  • Posts: 1212
Re: paraffin sectioning of mouse eyes
« Reply #3 on: July 20, 2011, 01:44:47 PM »
What thickness did you cut for the brain sections?

Offline DCundiff

  • SuperTech
  • ****
  • Posts: 23
Re: paraffin sectioning of mouse eyes
« Reply #4 on: July 21, 2011, 12:48:21 PM »
I am cutting my sections at 5 microns.

Offline richard03

  • Administrator
  • GoldMember
  • *****
  • Posts: 1212
Re: paraffin sectioning of mouse eyes
« Reply #5 on: July 21, 2011, 03:48:05 PM »
With 5 um thick paraffin section baked overnight at 37 C should be sufficient. I think the slides you are using caused the problem. Try other brand. TruBond 380 adhesive slides are very good.

Re: paraffin sectioning of mouse eyes
« Reply #5 on: July 21, 2011, 03:48:05 PM »