Immunoelectromicroscopic localization of surface antigens by GINS technique

GINS is Immunoelectromicroscopic technique combined indirect immunolabeling technique and negative staining technique in order to investigate surface antigens of particle specimens, such as virus, bacteria, debris of cell. This applicant investigation carry out on virus of JEV, HbsAg and salmonella. The results proved that samples not only have complete structure but also surface antigens with high specificity

Immuno-electronmicroscopic study on the serotoninergic taste cells and calcitonin gene-related peptide nerve fibers in mouse taste buds / 대한해부학회지

To investigate the distribution, ultrastructure and synapsis of serotoninergic cells and CGRP nerve fibers in mammalian taste buds, immunohistochemistry and electronmicroscopy were applied to mice vallate papillae. In normal mice, 1~2 serotonin immunoreactive cells were present in each taste bud section. After preloading 5-HTP, 3~6 cells showed strong immunoreactivity for serotonin. These cells were elongated, and their cytoplasm extended from the taste pore to the base of the taste bud. CGRP nerve fibers formed thick subgemmal nerve plexus under the basal lamina, and branched varicose perigemmal and intragemmal nerve fibers. Under the electron-microscope, three types of taste cells; dark cell, light cell and basal cells, were identified by their shape, location and electrical densities. Immuno-electronmicroscopy revealed that serotoninergic cells were dark cells. CGRP nerve fibers were located in and around taste buds, but the synaptic contacts with taste cells was not found. These findings indicate that mice taste cells are consisted of dark cell, light cell and basal cells, and dark cells contain serotonin. And, CGRP nerve fibers in taste buds may function as general sensory fibers.

COMPARATIVE STUDY OF TWO IMMUNOELECTRON MICROSCOPICAL DOUBLE LABELLING METHODS APPLYING PROTEIN A-GOLD TECHNIQUE / 解剖学报

In the present study, comparison have been made between two kinds of double labelling methods; single side labelling and both sides labelling. To make the different groups be comparable, all sections were continuously cut from the same specimen and all labelling processes were carried out at the same ti- me. The groups were divided as follows: A, one side anti-?-amylase labelling and one side antitrypsin labelling; B, single side labelling of anti-?-amylase and antitrypsin; C, one incubation with anti-?-amylase, followed by two incu- bations with protein A-gold (PAG) complexes of varied size, C1: 7nm and 20nm, C2: 10nm and 20nm, C3: 20nm and 7nm; D, single side labelling of anti- ?-amylase and an unrelated antiserum (antichathepsin D), applying free protein A between two labellings; E, as C, but with free protein A between two PAG incubations; F, as control. Group A and B showed that the two labelling me- thods had almost the same sensitivity.Group C indicated that the interaction of single side labelling were resulted from the combination of the second PAG with the free Fc region of the first antiserum. To decrease the interaction, it was necessary for the second PAG to be much larger than the first one.Group D de- monstrated that the interaction between the second antiserum and the first PAG was very feeble. Group E proved that free protein A could completely prevent the interaction of single side labelling method. The both sides labelling method avoids interaction, but mistakes resulting from the ultrastructural differences on two sides of the sections may happen. Which method to be selected is dependent upon what to be labelled.