The purpose of this report was to study the use of pre-embedding immunoelectron microscopy technique with gold and horseradish peroxidase (HRP) labeling in detecting the expression of ubiquitin C-terminal hydrolase 1 (UCH-L1) of podocytes in glomerulonephritis. The specimens of human IgA nephropathy and lupus nephritis were fixed with paraformaldehyde and lysine-HCl buffer, labeled by colloidal gold or HRP, embedded with epoxy resin, and examined under the transmission electron microscope. The high density of gold particles or peroxidase reaction products (DAB) combined with UCH-L1 was obvious in cytoplasm and processes of podocytes. This modified technique of pre-embedding immunoelectron microscopy could perfectly preserve the ultrastructure of kidney and expose antigens which is valuable for clinical diagnostic work and experimental research.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1080/01913120701829285 | DOI Listing |
J Vis Exp
July 2024
State Key Laboratory of Ophthalmology, Optometry and Visual Science, Eye Hospital, Wenzhou Medical University; Laboratory of Retinal Physiology and Disease, School of Ophthalmology and Optometry, Wenzhou Medical University;
The retina comprises numerous cells forming diverse neuronal circuits, which constitute the first stage of the visual pathway. Each circuit is characterized by unique features and distinct neurotransmitters, determining its role and functional significance. Given the intricate cell types within its structure, the complexity of neuronal circuits in the retina poses challenges for exploration.
View Article and Find Full Text PDFMicrosc Microanal
September 2023
Department of Physiology and Neurobiology, University of Connecticut, 75 North Eagleville Rd. Unit 3156, Storrs, CT 06269-3156, USA.
Electron microscopy (EM) volume reconstruction is a powerful tool for investigating the fundamental structure of brain circuits, but the full potential of this technique is limited by the difficulty of integrating molecular information. High quality ultrastructural preservation is necessary for EM reconstruction, and intact, highly contrasted cell membranes are essential for following small neuronal processes through serial sections. Unfortunately, the antibody labeling methods used to identify most endogenous molecules result in compromised morphology, especially of membranes.
View Article and Find Full Text PDFMethods Cell Biol
July 2023
Leon H Charney Division of Cardiology, Department of Medicine, New York University Grossman School of Medicine, New York, NY, United States.
Recent advances in volume electron microscopy (vEM) allow unprecedented visualization of the electron-dense structures of cells, tissues and model organisms at nanometric resolution in three dimensions (3D). Light-based microscopy has been widely used for specific localization of proteins; however, it is restricted by the diffraction limit of light, and lacks the ability to identify underlying structures. Here, we describe a protocol for ultrastructural detection, in three dimensions, of a protein (Connexin 43) expressed in the intercalated disc region of adult murine heart.
View Article and Find Full Text PDFJ Comp Neurol
August 2023
Laboratory of Retinal Physiology and Disease, School of Ophthalmology and Optometry and Eye Hospital, Wenzhou Medical University, Wenzhou, China.
The light pathways are segregated into rod and cone pathways in which rods synapse with rod bipolar cells (RBCs), while cones contact cone bipolar cells (CBCs). However, previous studies found that cones can make synapse with RBCs (cone-RBC synapses) and rods can contact OFF CBC in primate and rabbit retinas. Recently, such cone-RBC synapses have been reported physiologically and morphologically in the mouse retina.
View Article and Find Full Text PDFMethods Mol Biol
December 2022
Electron Microscopy Unit, Institute of Biotechnology, Helsinki Institute of Life Science, University of Helsinki, Helsinki, Finland.
Here, we describe protocols for chemical fixation and flat embedding to study the Golgi structure by thin section transmission electron microscopy (TEM) and for 3,3'-diaminobenzidine (DAB) cytochemical staining and pre-embedding immunolabelling to localize specific Golgi proteins. Furthermore, we demonstrate how the Golgi morphology can be elucidated by classifying the Golgi membranes using Microscopy Image Browser-a software that provides anonymization, modelling, and annotation.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!