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Demonstration of intracellular glycogen, peroxidases and viruses by light and electron microscopy
Author(s)
Tiedt, Friedrich A.C.
Date Issued
1995
Type
Thesis
Publisher
Cape Technikon
Abstract
The objective of the present study vvas primarily to determine
vvhether modifications to the existing osmium
tetroxide/potassium ferrocyanide method could be used as a
general ultrastructural glycogen stain and whether this
glycogen, vvhen so contrasted, differed in appearance in
different tumours, such as Evving's sarcoma,
Leiomyosarcoma, Rhabdomyosarcoma and Hepatocellular
carcinoma. Normal skeletal muscle and liver were used to
obtain a standard for the appearance of glycogen, and then,
with diagnostic criteria in mind, the four glycogen-rich
tumours, mentioned above vvere examined to determine the
appearance, distribution and amount of glycogen present in
them.
Modifications to the osmium tetroxide/potassium
ferrocyanide (OPF) method consisted of raising the
concentration of the ferrocyanide, and using no en bloc or
thin section staining by any uranyl salt solutions, because
solutions of uranyl salts leaches glycogen from tissue. This
procedure resulted in extremely electron-dense intracellular
glycogen being retained, vvhich aided diagnosis of the these
tumours. At high magnification. (>X30000) there vvas no
morphological difference betvveen the glycogen particles in
the various tumours, so these particles as such could not be
used as a diagnostic criterion.
Existing methods for the demonstration of myeloperoxidase
and platelet peroxidase vvere modified to obtain more precise
localization of the diaminobenzidine (DAB) reaction product.
Different anti-coagulants, one of which was heparin and a
modified fixation procedure vvas follovved.
vvhether modifications to the existing osmium
tetroxide/potassium ferrocyanide method could be used as a
general ultrastructural glycogen stain and whether this
glycogen, vvhen so contrasted, differed in appearance in
different tumours, such as Evving's sarcoma,
Leiomyosarcoma, Rhabdomyosarcoma and Hepatocellular
carcinoma. Normal skeletal muscle and liver were used to
obtain a standard for the appearance of glycogen, and then,
with diagnostic criteria in mind, the four glycogen-rich
tumours, mentioned above vvere examined to determine the
appearance, distribution and amount of glycogen present in
them.
Modifications to the osmium tetroxide/potassium
ferrocyanide (OPF) method consisted of raising the
concentration of the ferrocyanide, and using no en bloc or
thin section staining by any uranyl salt solutions, because
solutions of uranyl salts leaches glycogen from tissue. This
procedure resulted in extremely electron-dense intracellular
glycogen being retained, vvhich aided diagnosis of the these
tumours. At high magnification. (>X30000) there vvas no
morphological difference betvveen the glycogen particles in
the various tumours, so these particles as such could not be
used as a diagnostic criterion.
Existing methods for the demonstration of myeloperoxidase
and platelet peroxidase vvere modified to obtain more precise
localization of the diaminobenzidine (DAB) reaction product.
Different anti-coagulants, one of which was heparin and a
modified fixation procedure vvas follovved.
Additional information
Thesis (Masters Diploma (Medical Technology)--Cape Technikon, Cape Town, 1995
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