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Immunocytochemistry By Electron Spectroscopic Imaging Using Well Defined Boronated Monovalent Antibody Fragments, M. M. Kessels, B. Qualmann, W. D. Sierralta

Scanning Microscopy

Contributing to the rapidly developing field of immunoelectron microscopy a new kind of markers has been created. The element boron, incorporated as very stable carborane clusters into different kinds of peptides, served as a marker detectable by electron spectroscopic imaging (ESI) - an electron microscopic technique with high-resolution potential.

Covalently linked immunoreagents conspicuous by the small size of both antigen recognizing part and marker moiety are accessible by using peptide concepts for label construction and their conjugation with Fab' fragments. Due to a specific labeling of the free thiol groups of the Fab' fragments, the antigen binding capacity was not …

Labeling With Nanogold And Undecagold: Techniques And Results, James F. Hainfeld

Scanning Microscopy

A significant new development in gold labeling for microscopy has been achieved through the use of gold cluster compounds that are covalently attached to antibodies or other probe molecules. These unique gold probes are smaller than most colloidal gold conjugates and exhibit improved penetration into tissues, higher labeling densities, and allow many new probes to be made with peptides, nucleic acids, lipids, drugs, and other molecules. A new fluorescent-gold conjugate is useful for examining localization by fluorescence microscopy, then visualizing the same label at the ultrastructural level in the electron microscope.

Pre-Embedding Staining Of Single Muscle Fibers For Light And Electron Microscopy Studies Of Subcellular Organization, Evelyn Ralston, Thorkil Ploug

Scanning Microscopy

Skeletal muscle fibers are large, multinucleated cells which pose a challenge to the morphologist. In the course of studies of the distribution of the glucose transporter GLUT4, in muscle, we have compared different preparative procedures, for both light (LM) and electron microscopy (EM) immunocytochemistry. Here we show that pre-embedding staining of single teased fibers, or of single enzymatically dissociated fibers, has several advantages over the use of sections for observing discrete patterns that extend over long distances in the cells. We report on an optimization study carried out to establish fixation and permeabilization conditions for EM immunogold labeling of the …

Problems In Preparation Of Chromosomes For Scanning Electron Microscopy To Reveal Morphology And To Permit Immunocytochemistry Of Sensitive Antigens, A. T. Sumner

Scanning Microscopy

Although much information about chromosome structure and behaviour has been obtained using light microscopy, greater resolution is needed for a thorough understanding of chromosome organisation. Scanning electron microscopy (SEM) can provide valuable data about these three-dimensional organelles. The introduction of methods using osmium impregnation of methanol-acetic acid-fixed chromosome spreads revolutionised matters, producing life-like images of chromosomes. Nevertheless, it became clear that osmium impregnation introduced various artefacts, although the resulting images were still useful. Methanol-acetic acid-fixed chromosomes are, in fact, flattened on the glass substratum, and the 3-dimensional appearance obtained after osmium impregnation is the result of swelling during this process. …

Pathological And Immunocytochemical Changes In Chronic Calcium Oxalate Nephrolithiasis In The Rat, R. De Water, E. R. Boeve, P. P. M. C. Van Miert, C. P. Vermaire, P. R. W. A. Van Run, L. C. Cao, W. C. De Bruijn, F. H. Schroder

Scanning Microscopy

In the present study, we exposed rats to a crystal-inducing diet (CID) consisting of vitamin D3 and 0.5% ethylene glycol (EG), and we investigated histologically the kidney damage induced by the deposition of calcium oxalate (CaOx) crystals. After 28 days, 50 % of the animals had renal CaOx crystals, of which 60% also had small papillary stones. Most crystals were present in the cortex. The occurrence of these crystals coincided with morphological and cytochemical changes: glomerular damage, tubular dilatation and necrosis, and an enlargement of the interstitium. The number of epithelial and interstitial cells positive for the proliferating cell nuclear …