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Introduction Fixation
o Fixative should be at 4C for
Enzymes are large tertiary the shortest time.
proteins that acts as catalyst and are Formol Calcium
vital in cellular metabolism while o Recommended for tissue
maintaining hemostasis in man. blocks
Histochemistry is defined as the o Help maintain cell membrane
identification, localization and integrity
qualification in cells and tissue and o Used on smears
by chemical or physical test of o Cryostat Sections
specific substances, reactive groups Buffered Formalin or Formal
and enzyme catalyzed substance. Saline
Enzyme Histochemistry o Satisfactory in most cases
(commonly used)
This technique produces a final Acetone, Formalin Vapour and
coloured reaction product that is Formalin-Alcohol Mixtures
localized to the cells with an intensity
proportional to the enzyme activity.
Applying on enzyme detection Principle of Enzyme Histochemistry
system to tissue section or smears.
Histochemistry procedures are
Classification based on the simple premise that
tissue cells, when placed in a
Classified according to their effect
solution chemically react with the
on the substrate:
solution to produce a colored
o Oxidoreductases
insoluble product.
The most important
groups to the The amount and location of the end-
diagnostic enzyme product can then be evaluated in the
histochemist. context of the cells or tissue.
Known as oxidases Classical histochemical reaction and
and dehydrogenases. generally based on one of the FOUR
o Transferases PRINCIPLES:
o Hydrolases
Oxidative and 1. Simple ionic interactions
hydrolytic enzymes 2. Reactions of aldehydes with
o Lyases schiffs reagent or silver
o Isomerases compounds
o Ligases 3. Coupling of aromatic
diazonium salts with aromatic
Activators residues on protein
4. Conversion acting on a
Ions of Calcium
substrate to form colored ppt.
Magnesium
Manganese
Sodium
Potassium
Types of Histochemical Reactions Self-coulered Substrate