[Histonet] Re: embedding problem small zebrafish
TJJ <@t> Stowers-Institute.org
Tue Mar 25 15:24:39 CDT 2008
Barbara is using Technovit 9100 which is a methyl methacrylate resin, I'm presuming because she is not using any decalcification methods on her fish.
We have not used this for any aquatic species, but have done some work with femurs and spinal columns. The trick for getting good sections is to wet the block face with 30% alcohol before you cut it. We used a D-profile tungsten carbide knife on a retracting electronic microtome (Leica or Microm). The website for the Technovit 9100 kit (http://www.emsdiasum.com/microscopy/technical/datasheet/14655.aspx) has these instructions:
* The blocks must be trimmed before cutting
* Use of 16 mm hardened metal knife with D-form cutting edge or HKS-Knives. When cutting polymerized Technovit 9100 NEW blocks, 30% ethanol (cutting fluid) must be used
* Transfer sections to super frost plus slides, mount with 50% ethanol (mounting fluid) and cover with PVC-foil (Kisol-foil)
* Remove excess fluid with filter paper. Load the slides into a section-press
We made the press ourselves using wooden tongue depressors and a clamp. Each slide is covered with regular plastic wrap, and then they are all stacked together with the tongue depressors on the top and bottom of the stack. They are then clamped and placed in a 50 degree oven overnight.
Is this what you are currently doing?
Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110
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