[Histonet] Block staining with HE

[John Kiernan]

You need to read the paper. It says "All routinely used hematoxylins, such as Ehrlich’s, Mayer’s, Harris’ and Delafield’s, proved unsatisfactory"

STAIN TECHNOLOGY
Vol. 56, No. 2, pp. 119-123 (1981)
BLOCK STAINING OF MAMMALIAN TISSUES WITH HEMATOXYLIN AND EOSIN
IAN F. HINE, Department of Anatomy, Monash University, Clayton, Victoria 3168, Australia.

ABSTRACT. Various mammalian tissues were stained en bloc with hematoxylin and eosin after fixation and prior to embedding in paraffin wax and sectioning. The choice of fixative is important and best results are obtained using Worcester’s Fluid, a combination of saturated aqueous mercuric chloride, formaldehyde, and glacial acetic acid. After fixation, blocks of tissue up to 1.5 em thick are stained for seven days in hematoxylin. Excess stain is removed by washing tissues in running water overnight. Tissue blocks then are dehydrated with graded concentrations of ethyl alcohols to 80% and counterstained, with further dehydration, in 0.5% spirit soluble eosin in 90% ethyl alcohol for five
days. The tissue is subsequently transferred to 90% ethyl alcohol overnight to differentiate win staining; dehydration is completed in absolute ethyl alcohol. The blocks are cleared in cedarwood oil and briefly in xylene prior to embedding, sectioning, and mounting. Following removal of wax by xylene, coverslips are applied. General morphological and histological features were particularly well differentiated and very selectively and reliably stained by this method.

Hine's "Worcester's" fixative is similar to Stieve's. His haemalum formulation is a little unusual. The counterstain was ethyl eosin (CI 45385), not eosin Y (CI 45380). There is a page of colour photos in which the red/blue colour separation is satisfactory in only two (my opinion). One photo shows a block-stained H&E section subsequently stained with Masson's and Verhoeff's, with bluish green collagen and black elastic laminae.

John Kiernan
London, Canada
= = =
On 20/03/15, Caroline Miller  <mills <@t> 3scan.com> wrote:
> Hi Histonet, Happy Friday!
> 
> I was wondering if anyone had access to this paper:
> Stain Technol.
> <http://www-ncbi-nlm-nih-gov.ucsf.idm.oclc.org/pubmed/6166093?dopt=AbstractPlus&tool=cdl&otool=cdlotool#>
> 1981 Mar;56(2):119-23.
> Block staining of mammalian tissues with hematoxylin and eosin.
> 
> or if anyone had some tips on whole-mount HE staining I would gladly take
> it!
> 
> I am just starting on this quest, but so far I have tried vacuum for the
> haematoxylin overnight with Erlichs and Harris's, and I have some Mayers on
> order. The Erlichs and harris's really overstained the tissue and I have
> been destaining for a while. I am thinking Mayer's may be better for its
> progressive nature, but any advice will be gratefully taken!
> 
> Thank you in advance for your fantastic advice!
> 
> yours,
> 
> Caroline
> 
> 
> 
> 
> -- 
> Caroline Miller
> Director of Histology
> 3Scan.com
> 415 2187297
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