SYNAPTOPHYSIN - AN IMMUNO-HISTOCHEMICAL MARKER FOR CHILDHOOD NEUROBLASTOMA

A new cytochemical method is presented for the light and electron microscopic localization of lysosomes in mineralized and soft tissues. Inorganic trimetaphosphate is used as substrate in a lead chelate incubation medium at pH 3.9. Lysosomes in several tissues are strongly reactive, and reaction product is frequently present in Golgi saccules and GERL. The reaction can be differentiated from acid glycerophosphatase activity, is relatively insensitive to fixation and demineralization procedures, and the reaction is often complete after short incubation times.

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Earliest Steps in Primary Tumor Formation and Micrometastasis Resolved with Histochemical Marker Gene-Tagged Tumor Cells

Microscopy and Microanalysis ◽

10.1017/s1431927600007042 ◽

1997 ◽

Vol 3 (S2) ◽

pp. 31-32

Author(s):

L. A. Culp ◽

W.-c. Lin ◽

N. Kleinman ◽

K. O'Connor ◽

R. Lechner

Keyword(s):

Tumor Cells ◽

Marker Gene ◽

Cell Number ◽

Athymic Nude Mouse ◽

Tumor Formation ◽

Placental Alkaline Phosphatase ◽

3T3 Cells ◽

Nude Mouse Model ◽

Human Placental Alkaline Phosphatase ◽

Histochemical Marker

To facilitate detection of tumor cells at the highest resolution in an athymic nude mouse model system, Balb/c 3T3 cells transformed with the Harvey ras oncogene were transfected with the histochemical marker gene, bacterial lacZ (LZEJ cells). Alternatively, 3T3 cells transformed with the human sis oncogene were transfected with human placental alkaline phosphatase marker gene (APSI cells). Within minutes of subcutaneous injection, these tumor cells could be detected histochemically and the fate of cells followed with time. APSI or LZEJ cells gave very different single-cell morphologies at this site but yielded similar aggregation patterns of cells. Clearance of some cells could readily be detected by diffusion of histochemical product as “curly-haired” organizations of cells condensed into ovoid collections. Expansion of the population occurred with division of many cells in the population, not just one or a few cells. Cell number was quantitated by the development of ultrasensitive luminometry assays for the two histochemical marker enzymes; different kinetics were observed for establishment of LZEJ or APSI cells.

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Na+K+ Dependent Atpase as a Specific Marker for Myoepithelial Cells: An Ultrastructural Study.

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100115894 ◽

1975 ◽

Vol 33 ◽

pp. 454-455

Author(s):

J. Russo ◽

P. Wells

Keyword(s):

Osmium Tetroxide ◽

Plasma Membranes ◽

Mammary Glands ◽

Myoepithelial Cells ◽

Cellular Heterogeneity ◽

Specific Marker ◽

Dependent Atpase ◽

Cacodylate Buffer ◽

Histochemical Marker ◽

Cellular Types

One of the problems in the study of mammary carcinomas is the cellular heterogeneity which they present. The use of a specific histochemical marker can help to distinguish the two principal cellular types found in the mammary gland; the epithelial and myoepithelial cells.Mammary glands of female Balb/C mice of different ages were removed and fixed in 1.5% glutaraldehyde in 0.1M cacodylate buffer pH 7.2 with 1% sucrose. After two hours of fixation at 4°C, the material was washed in 0.1M cacodylate buffer, sectioned with a Smith-Farquar microtome set at 70-100u, and incubated for the histochemical detection of ATPases. The tissue was then washed again with 0.1M cacodylate buffer, postfixed in 1% osmium tetroxide, dehydrated, and embedded in an Epon-Araldite mixture.For the Mg++ dependent ATPase the conventional method of Wachstein and Meisel was followed. The Mg++ dependent ATPase is localized in the plasma membranes of the epithelial and myoepithelial cells (Fig. 1).

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Demonstration of tartrate-resistant acid phosphatase in un-decalcified, glycolmethacrylate-embedded mouse bone: a possible marker for (pre)osteoclast identification.

Journal of Histochemistry & Cytochemistry ◽

10.1177/34.10.3745910 ◽

1986 ◽

Vol 34 (10) ◽

pp. 1317-1323 ◽

Cited By ~ 144

Author(s):

F P van de Wijngaert ◽

E H Burger

Keyword(s):

Bone Marrow ◽

Acid Phosphatase ◽

Phosphatase Activity ◽

Acid Phosphatase Activity ◽

Mononuclear Cells ◽

Histochemical Demonstration ◽

Splenic Tissue ◽

Tartrate Resistant Acid Phosphatase ◽

Excellent Preservation ◽

Histochemical Marker

Fixed, undecalcified mouse long bones were embedded in glycol methacrylate (GMA), sectioned, and incubated for acid phosphatase in the presence or absence of tartrate, to investigate the feasibility of tartrate-resistant acid phosphatase as a histochemical marker for osteoclast identification. Naphthol AS-BI phosphate was used as the substrate and hexazonium pararosanaline as coupler. Cytocentrifuge preparations of mouse, rat, and quail bone marrow or frozen and GMA sections of mouse splenic tissue were used as controls to specify acid phosphatase activity. After adequate fixation, acid phosphatase activity sensitive to tartrate inhibition (TS-AP) was demonstrated in macrophages from spleen, bone marrow, and loose connective tissue surrounding bone rudiments. Acid phosphatase activity resistant to tartrate inhibition (TR-AP), was detected in multi-nuclear osteoclasts and in some mononuclear cells from bone marrow and periosteum. In cytocentrifuge preparations and frozen sections of mouse spleen, TR-AP was demonstrated after simultaneous incubation with substrate and tartrate. In GMA sections, however, TR-AP could only be demonstrated after pre-incubation with tartrate before application of substrate. We suggest that histochemical demonstration of TR-AP versus TS-AP on GMA-embedded bone sections by means of a pre-incubation method can be used as an identification marker of (pre)osteoclasts. Plastic embedding is recommended for its excellent preservation of morphology and enzyme activity.

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