Timm’s sulphide silver method has so far
been considered a very sensitive technique for demonstrating metal ions, in
particular zinc in the central nervous system. The principle of this technique
is that metals in the tissue can be transformed histochemically to metal
sulphides. Subsequently, metal sulphides catalyze the reduction of silver ions
by a reducing agent to metallic grains that are visible under a light or
electron microscope (for review, cf. ref. 1). Studies using this technique have
provided a better understanding of not only the localization and distribution
of zinc but also its possible function in the brain1. Nevertheless, the reliability and complexity of Timm's sulphide silver stains often act as a limiting factor to its widespread applications for the studies of neurodegenerative diseases.
FD Rapid TimmStain Kit™ is designed based
on the methods described by Haug1and Sloviter2. The reagents and
procedure of the FD Rapid TimmStain Kit™ have been optimized to achieve a high
degree of both specificity and sensitivity for the demonstration of metal ions,
especially zinc in the brain of experimental animals. The kit can be used with
both frozen and paraffin sections. Moreover, the Timm-stained sections can be counterstained with nuclear stains, such as Nissl to visualize more details of seizure-induced morphological alterations (see below).
contents:
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Part 1 (stored at 4°C):
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Perfusate A
500 ml
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Perfusate B
500 ml
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Part 2 (stored at -20°C):
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Solution A
220 ml
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Solution B
200 ml
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Solution C
200 ml
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Solution D
3 ml
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Plastic forcepts & User Manual
1 of each
Materials Required, but Not Included:
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Double distilled or deionized water
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10% neutral buffered formalin
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0.1 M phosphate buffer (pH7.4)
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Histological supplies and equipment: Microscopic slides, coverslips, staining jars, xylene or xylene substitutes, resinous mounting medium (e.g. Permount™) and a light microscope
References:
1.
Haug F-M S (1973) Heavy
metals in the brain: a light microscope study of the rat with Timm’s sulphide
silver method. Methodological considerations and cytological and regional
staining patterns.
Springer-VerlagBerlinHeidelbergNew York.
2.
Sloviter RS (1982) A
simplified Timm stain procedure compatible with formaldehyde fixation and
routine paraffin embedding of rat brain. Brain Research Bulletin 8:771-774.