Ultrastructural Study of Glomerular Capillary Loops at Different Perf usion Pressures as Revealed by Quick-Freezing, Freeze-Substitution and Conventional Fixation Methods

Nephron ◽  
1997 ◽  
Vol 76 (4) ◽  
pp. 452-459 ◽  
Author(s):  
Ying Yu ◽  
Chong-Guang Leng ◽  
Yasuko Kato ◽  
Shinichi Ohno
Keyword(s):  
Ultrastructural Study ◽  
Freeze Substitution ◽  
Fixation Methods ◽  
Glomerular Capillary ◽  
Quick Freezing

How to sample the entire length of a single muscle fiber quick-frozen after electrical point stimulation for high resolution EM

1992 ◽  
Vol 50 (1) ◽  
pp. 776-777
Author(s):  
Joachim R. Sommer ◽  
Teresa High ◽  
Betty Scherer ◽  
Isaiah Taylor ◽  
Rashid Nassar
Keyword(s):  
Skeletal Muscle ◽  
High Resolution ◽  
Action Potential ◽  
Muscle Fiber ◽  
Freeze Fracture ◽  
Freeze Substitution ◽  
Electrical Field Stimulation ◽  
Skeletal Muscle Fiber ◽  
Freeze Dried ◽  
Quick Freezing

We have developed a model that allows the quick-freezing at known time intervals following electrical field stimulation of a single, intact frog skeletal muscle fiber isolated by sharp dissection. The preparation is used for studying high resolution morphology by freeze-substitution and freeze-fracture and for electron probe x-ray microanlysis of sudden calcium displacement from intracellular stores in freeze-dried cryosections, all in the same fiber. We now show the feasibility and instrumentation of new methodology for stimulating a single, intact skeletal muscle fiber at a point resulting in the propagation of an action potential, followed by quick-freezing with sub-millisecond temporal resolution after electrical stimulation, followed by multiple sampling of the frozen muscle fiber for freeze-substitution, freeze-fracture (not shown) and cryosectionmg. This model, at once serving as its own control and obviating consideration of variances between different fibers, frogs etc., is useful to investigate structural and topochemical alterations occurring in the wake of an action potential.

Ultrastructural study of cytoskeletal interactions with endosomes in macrophages by quickfreezing and deep-etching method

1990 ◽  
Vol 48 (3) ◽  
pp. 576-577
Author(s):  
Takeshi Baba ◽  
Nobuki Shiozawa ◽  
Masao Hotch ◽  
Shinichi Ohno
Keyword(s):  
Immune Complex ◽  
Ultrastructural Study ◽  
Fc Receptor ◽  
Coated Pits ◽  
Deep Etching ◽  
Receptor Mediated Endocytosis ◽  
Etching Method ◽  
Quick Freezing

Endosomes are vesicular or tubular organelles that play important roles in transports of receptors and receptor―bound ligands during receptor-mediated endocytosis. The mechanisms of endocytic transports from clathrin-coated pits to lysosomes have been studied by many investigators. However, few studies were reported about the interactions between endosomes and cytoskeletons. In this study, Fc-receptor-mediated endocytosis in macrophages are investigated by quick-freezing and deep-etching (QF-DE) method combined with gold-labeled immune complex and “replica scraping method”.

Ultrastructural Study of the Cytoskeleton of Optic Nerve Axons in Guinea Pigs as Revealed by a Quick-Freezing, Deep-Etching Method

1996 ◽  
Vol 28 (1) ◽  
pp. 29-35
Author(s):  
Bo Ou ◽  
Shinichi Ohno ◽  
Nobuo Terada ◽  
Yasuhisa Fujii ◽  
Hai-Bo Chen ◽  
...  
Keyword(s):  
Optic Nerve ◽  
Ultrastructural Study ◽  
Guinea Pigs ◽  
Deep Etching ◽  
Etching Method ◽  
Quick Freezing

Ultrastructural study of the fission yeast Schizosaccharomyces pombe nucleolus by freeze substitution

1995 ◽  
Vol 84 (3) ◽  
pp. 228-228
Author(s):  
Leger Isabelle ◽  
Jacqueline Noaillac-Depeyre ◽  
Marie-Pierre Gulli ◽  
Michèle Caizergues-Ferrer ◽  
Nicole Gas
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Ultrastructural Study ◽  
Freeze Substitution

Ultrastructural study ofCryptococcus neoformans by quick-freezing and deep-etching method

1993 ◽  
Vol 121 (3) ◽  
pp. 133-141 ◽  
Author(s):  
Nobuki Sakaguchi ◽  
Takeshi Baba ◽  
Masao Fukuzawa ◽  
Shinichi Ohno
Keyword(s):  
Ultrastructural Study ◽  
Deep Etching ◽  
Etching Method ◽  
Quick Freezing

Quick-Freezing to Catch the Membrane Changes that Occur During Exocytosis

1977 ◽  
Vol 35 ◽  
pp. 676-679
Author(s):  
J.E. Heuser
Keyword(s):  
Synaptic Vesicle ◽  
Structural Changes ◽  
Freeze Fracture ◽  
Freeze Substitution ◽  
Frog Neuromuscular Junction ◽  
The Past ◽  
Synaptic Vesicle Exocytosis ◽  
Quick Freezing ◽  
Vesicle Exocytosis ◽  
Membrane Changes

The technique that we have used to capture synaptic vesicle exocytosis at the frog neuromuscular junction - that of quick-freezing muscles followed by freeze fracture (3) or freeze substitution (6) - works sufficiently well now that it may be useful in other sorts of membrane studies, or studies of fast structural changes with the electron microscope. This note reviews the quickfreezing technique we use, and describes its application to the problem of synaptic vesicle exocytosis and recycling at the synapse.Here, many of the membrane changes of interest occur during the brief delay in synaptic transmission, on a time scale of milliseconds or fractions of milliseconds, and leave only traces thereafter. In the past, we have studied these left-over traces in tissues fixed with the standard chemicals for electron microscopy (1), and have inferred from them that vesicles discharge the quanta of neurotransmitters, as the physiologists would predict.

Ultrastructural Study of Peroxisome Proliferation in Hepatocytes by Quick-Freezing and Deep-Etching Method

1988 ◽  
Vol 46 ◽  
pp. 258-259
Author(s):  
S. Ohno
Keyword(s):  
Electron Microscopy ◽  
Liquid Nitrogen ◽  
Ultrastructural Study ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferation ◽  
Solution Ph ◽  
Deep Etching ◽  
Quick Freezing ◽  
Ethylhexyl Phthalate ◽  
Conventional Electron Microscopy

It had been emphasized that luminar continuities between sER and peroxisomes were detected by conventional electron microscopy. However, recent studies ruled out the luminar continuities between sER and peroxisomes. Lazarow et al. reported that peroxisomal proteins were synthesized on free ribosomes and postulated the existence of “peroxisomal reticulum” distinct from the ER. The object of this study is to clarify the proliteration mechanism of peroxisomes after administration of a peroxisome proliferator, DEHP (di-2- ethylhexyl phthalate).Mice treated with 2% DEHP for 1, 3, 5 and 7 days and normal mice were perfused with 2% paraformaldehyde in 0.1M phosphate buffered solution, pH 7.4, (PB) for 5 min. The livers were cut into small pieces, washed in PB to remove cytoplasmic soluble proteins and were fixed again with 2% paraformaldehyde-0.25% glutaraldehyde for 30 min. They were quickly frozen in isopentane-propane mixture (around -190 C) and fractured in liquid nitrogen to remove the damaged surface tissues. They were deeply etched in Eiko FD-3S machine (-95°C, 2-6xl0-7 Torr) and rotary shadowed with platinum.

Immunocytochemical localization of xanthine oxidase in rat hepatocytes processed by quick freezing followed by freeze-substitution fixation

1990 ◽  
Vol 48 (3) ◽  
pp. 908-909
Author(s):  
Misao Ichikawa ◽  
Atsushi Ichikawa ◽  
Takeshi Nishino ◽  
Tomako Nishino
Keyword(s):  
Xanthine Oxidase ◽  
Freeze Substitution ◽  
Rat Hepatocytes ◽  
Electron Microscopic Level ◽  
Contact Method ◽  
Electron Microscopic ◽  
Rat Hepatocyte ◽  
Peroxisomal Membrane ◽  
Fresh Tissue ◽  
Quick Freezing

Quick freezing of tissues, with freeze-substitution fixation, not only provides superior ultrastructural detail in cells and tissues but also retains soluble substances within their normal intracellular compartments. Combining this procedure with postembedding protein A-gold immunocytochemical technique permits precise localization of antigenic sites at the electron microscopic level.Biochemical and cytochemical studies have demonstrated variously that xanthine oxidase, one of the flavoprotein enzymes involved in urate metabolism, is localized in cytozol, peroxisomal membrane, and/or peroxisomal crystalline cores of rat hepatocytes. This study is designed to determine the localization of xanthine oxidase in rat hepatocyte by high resolution immuno-electron microscopy.Adult Wistar rats, 150-230 g, maintained on standard rat food pellets and water ad libitum were fasted for 12 h before sacrifice. To examine the presence of xanthine oxidase in peroxisomes in rat hepatocyte, some animals were treated with intramuscular injections of 250 mg (per day) of clofibrate, a hypolipidemic drug, for 10 days to generate peroxisome proliferation. Fresh tissue blocks were quickly frozen by the metal contact method, using liquid helium, and then freeze-substituted with either 2% OsO4 in acetone or 0.3% glutaraldehyde-acetone solution.

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