Immunohistochemical Localization of Coelomic Fluid-Specific Protein in the Coelomic Epithelium and Mesovarium ofChum Salmon Oncorhynchus keta.

The nephridia of the adult Asymmetron are described for the first time. They are of the protonephridial type, and resemble those of Branchiostoma, though smaller and simpler in structure. The canal of the paired nephridia is in the shape of a triangular flattened sac into which open numerous solenocytes. The sac and the solenocytes extend over the inner wall of the suprapharyngeal coelom, and the external pore opens into the atrium near the top of the secondary bar. The first pair of nephridia, however, is situated in the epipterygial coelom. The paired nephridia are ‘retroperitoneal’ ; but the coelomic epithelium is interrupted, and does not cover the solenocyte field, so that coelomic fluid bathes the solenocyte tubes. The nephridium of Hatschek of Asymmetron resembles that of Branchiostoma, but the diverticula are shorter. The nephridia of the two genera are compared, and certain details in the structure of those of Branchiostoma are discussed.

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Densitometric Identification of Primitive Erythroblasts After Reaction With Diaminobenzidine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072083 ◽

1973 ◽

Vol 31 ◽

pp. 328-329

Author(s):

John A. Trotter

Keyword(s):

Red Blood Cells ◽

Analytical Method ◽

Blood Cells ◽

Guinea Pigs ◽

Specific Protein ◽

Visual Examination ◽

Hemoglobin Synthesis ◽

Peroxidatic Activity ◽

Small Density

Hemoglobin is the specific protein of red blood cells. Those cells in which hemoglobin synthesis is initiated are the earliest cells that can presently be considered to be committed to erythropoiesis. In order to identify such early cells electron microscopically, we have made use of the peroxidatic activity of hemoglobin by reacting the marrow of erythropoietically stimulated guinea pigs with diaminobenzidine (DAB). The reaction product appeared as a diffuse and amorphous electron opacity throughout the cytoplasm of reactive cells. The detection of small density increases of such a diffuse nature required an analytical method more sensitive and reliable than the visual examination of micrographs. A procedure was therefore devised for the evaluation of micrographs (negatives) with a densitometer (Weston Photographic Analyzer).

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Specific Labeling of Protein Domains with Antibody Fragments

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100098587 ◽

1981 ◽

Vol 39 ◽

pp. 416-417

Author(s):

U. Aebi ◽

L.E. Buhle ◽

W.E. Fowler

Keyword(s):

Image Processing ◽

Specific Protein ◽

Antibody Fragments ◽

Supramolecular Organization ◽

Two Dimensional ◽

Ordered Structures ◽

Virus Capsids ◽

Processing Techniques

Many important supramolecular structures such as filaments, microtubules, virus capsids and certain membrane proteins and bacterial cell walls exist as ordered polymers or two-dimensional crystalline arrays in vivo. In several instances it has been possible to induce soluble proteins to form ordered polymers or two-dimensional crystalline arrays in vitro. In both cases a combination of electron microscopy of negatively stained specimens with analog or digital image processing techniques has proven extremely useful for elucidating the molecular and supramolecular organization of the constituent proteins. However from the reconstructed stain exclusion patterns it is often difficult to identify distinct stain excluding regions with specific protein subunits. To this end it has been demonstrated that in some cases this ambiguity can be resolved by a combination of stoichiometric labeling of the ordered structures with subunit-specific antibody fragments (e.g. Fab) and image processing of the electron micrographs recorded from labeled and unlabeled structures.

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Immunocytochemical localization of p65 with one-nanometer gold particles following silver development

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010015719x ◽

1989 ◽

Vol 47 ◽

pp. 1042-1043

Author(s):

Richard W. Burry ◽

Diane M. Hayes

Keyword(s):

Plasma Membrane ◽

Bovine Serum Albumin ◽

Calf Serum ◽

Specific Protein ◽

Immunocytochemical Localization ◽

Electron Microscopic ◽

Gold Particles ◽

Pass Through ◽

Label Distribution ◽

Phosphate Buffered Saline

Electron microscopic (EM) immunocytochemistry localization of the neuron specific protein p65 could show which organelles contain this antigen. Antibodies (Ab) labeled with horseradish peroxidase (HRP) followed by chromogen development show a broad diffuse label distribution within cells and restricting identification of organelles. Particulate label (e.g. 10 nm colloidal gold) is highly desirable but not practical because penetration into cells requires destroying the plasma membrane. We report pre-embedding immunocytochemistry with a particulate marker, 1 nm gold, that will pass through membranes treated with saponin, a mild detergent.Cell cultures of the rat cerebellum were fixed in buffered 4% paraformaldehyde and 0.1% glutaraldehyde (Glut.). The buffer for all incubations and rinses was phosphate buffered saline with: 1% calf serum, 0.2% saponin, 0.1% gelatin, 50 mM glycine 1 mg/ml bovine serum albumin, and (not in the HRP labeled cultures) 0.02% sodium azide. The monoclonal #48 to p65 was used with three label systems: HRP, 1 nm avidin gold with IntenSE M development, and 1 nm avidin gold with Danscher development.

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