Spurr ? and TEM stain reply
Virginia, yes, our lab uses Spurr epoxy more information I invite you to
view our WEB PAGE
For instructions on how to handle renal biopsies and other biopsies, click
on V.DIAGNOSTIC SERVICES. Then click on renal biopsy, biopsy for HRLM-TEM,
muscle biopsy, nerve biopsy or other sub-titles. To better understand our
approach for the study and interpretation of the renal biopsies, click on
1. RENAL PATHOLOGY and then on GLOMERULOPATHIES or RENAL PATHOLOGY 2000.
OK, we TEM stain with Uranyl Acetate and Lead Citrate as follows:
URANYL ACETATE (UA)
Use sterile freshly boiled water (use while water is still hot!)
0.5 gm uranyl acetate
5ml hot, sterile boiled H20
Make in 15 ml conical tube. Vortex briefly. Add 20 microliters of glacial
acetic acid. Vortex for several seconds. Transfer to a 5ml syringe, Place
filter (Fisherbrand Cat No 09-7198B 0.45um Sterile, I am sure another brand
may be used, that is what our suppy dept has on hand) on end. check ph it
should be 4. Stain is good for 7 days. Wrap UA in foil to avoid light
exposure. Change filters every time you stain, as there is a build up on
tip that may cause contamination.
LEAD CITRATE (LC)
0.02 gm lead citrate
5ml Hot, sterile, boiled H20
Make in 15 ml conical tube. Vortex briefly. Add 20 microliters of 10N NaOH.
Vortex for several seconds. Transfer toa 5ml syringe. place filter on end
(Fisherbrand Cat No 09-7198B 0.45um Sterile, I am sure another brand may be
used, that is what our suppy dept has on hand). Check pH it should be 12.
Stain is good for 7 days. Wrap LC in foil to avoid light exposure. Change
filters every time you stain, as there is a build up on tip that may cause
Please keep in touch and let me know how its working for you, Teresa
>I found your email on histonet responding to a "Don't
>Use Spurr's" tirade. I've used it quite successfully
>in the past and am now having some trouble with the
>tissue I'm working with. Do you have a good protocol
>for staining for TEM? I understand methanolic UA works
>best but I'm having problems with tissue folding as
>well. Also, do you stain en bloc with UA? And which
>lead citrate do you like?
>Thanks for your help.
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