scholarly journals SOLUBLE HEMIN COMPOUNDS AS ULTRASTRUCTURAL TRACERS

1973 ◽  
Vol 21 (12) ◽  
pp. 1047-1052 ◽  
Author(s):  
J. F. ARONSON ◽  
A. P. FISHMAN ◽  
G. G. PIETRA
Keyword(s):  
Electron Microscope ◽  
Horseradish Peroxidase ◽  
Peroxidase Activity ◽  
Glucuronic Acid ◽  
Prosthetic Group ◽  
Electron Microscopic ◽  
Size Classes ◽  
Ultrastructural Studies ◽  
Microscopic Localization ◽  
Electron Microscopic Localization

We have demonstrated that soluble hemin derivatives have electron microscope-detectable peroxidase activity. Hemin was coupled to several different size classes of dextran, to sucrose and to glucuronic acid. The peroxidase activity of the attached hemin was detected with the electron microscope using the method described by Graham and Karnovsky for horseradish peroxidase. These observations indicate that hemin may be used as a prosthetic group to label macromolecules for electron microscopic localization and raise the prospect of creating groups of inexpensive tracer substances of varying size, charge and specificity for histologic and ultrastructural studies.

Pollen-Wall Proteins: Electron-Microscopic Localization of Acid Phosphatase in the Intine of Crocus Vernus

1971 ◽  
Vol 8 (3) ◽  
pp. 727-733
Author(s):  
R. B. KNOX ◽  
J. HESLOP-HARRISON
Keyword(s):  
Electron Microscope ◽  
Acid Phosphatase ◽  
Coupling Reaction ◽  
Pollen Grain ◽  
Microscope Level ◽  
Electron Microscopic ◽  
Electron Microscope Level ◽  
Microscopic Localization ◽  
Filamentous Inclusions ◽  
Electron Microscopic Localization

Acid phosphatase has been localized in the wall of the pollen grain of Crocus vernus Wulf at the electron-microscope level by a method using 2-naphthyl thiol phosphate as substrate in a simultaneous coupling reaction with fast blue BBN at pH 5.0, the product being given electron opacity by osmication. Activity was found to be concentrated mainly in the intine, and to be associated with ribbon-like or filamentous inclusions believed to be proteinaceous on the basis of other criteria. Some activity was also detectable in the interstices of the exine. The observations confirm the general interpretation of the distribution of wall-held enzyme based upon light-microscopic cytochemistry, and provide the resolution necessary to establish unambiguously that they are associated with protein layers inserted during intine growth.

The electron microscopic localization of nephrotoxic antibodies in isolated glomeruli

Pathology ◽  
1976 ◽  
Vol 8 (1) ◽  
pp. 73-80 ◽  
Author(s):  
I.P. McCausland ◽  
R.N. Seelye ◽  
J.B. Gavin ◽  
P.B. Herdson
Keyword(s):  
Electron Microscopic ◽  
Isolated Glomeruli ◽  
Microscopic Localization ◽  
Electron Microscopic Localization

scholarly journals An artificial test substrate for evaluating electron microscopic immunocytochemical labeling reactions.

1987 ◽  
Vol 35 (8) ◽  
pp. 909-916 ◽  
Author(s):  
G D Gagne ◽  
M F Miller
Keyword(s):  
Electron Microscope ◽  
Hepatitis B ◽  
Horseradish Peroxidase ◽  
Polyclonal Antibodies ◽  
Hepatitis B Surface Antigen ◽  
Immunogold Labeling ◽  
Artificial Substrate ◽  
Chemical Fixation ◽  
Electron Microscopic ◽  
Immunocytochemical Studies

We describe an artificial substrate system for optimization of labeling parameters in electron microscope immunocytochemical studies. The system involves use of blocks of glutaraldehyde-polymerized BSA into which a desired antigen is incorporated by a simple soaking procedure. The resulting antigen-impregnated artificial substrate can then be fixed and embedded identically to a piece of tissue. The BSA substrate can also be dried and then sectioned for immunolabeling with or without chemical fixation and without exposing the antigen to dehydrating agents and embedding resins. The effects of various fixation and embedding procedures can thus be evaluated separately. Other parameters affecting immunocytochemical labeling, such as antibody and conjugate concentration, can also be evaluated. We used this system, along with immunogold labeling, to determine quantitatively the optimal fixation and embedding conditions for labeling of hepatitis B surface antigen (HbsAg), human IgG, and horseradish peroxidase. Using unfixed and unembedded HBsAg, we were able to detect antigen concentrations below 20 micrograms/ml. We have shown that it is not possible to label HBsAg within resin-embedded cells using conventional aldehyde fixation protocols and polyclonal antibodies.

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