Detection of injected fluorescence-conjugated IgG in living mouse organs using “in vivo cryotechnique” with freeze-substitution

2005 ◽  
Vol 66 (4) ◽  
pp. 173-178 ◽  
Author(s):  
Nobuo Terada ◽  
Nobuhiko Ohno ◽  
Zilong Li ◽  
Yasuhisa Fujii ◽  
Takeshi Baba ◽  
...  
Keyword(s):  
Freeze Substitution ◽  
In Vivo Cryotechnique ◽  
Living Mouse

scholarly journals Dynamic ultrastructure of mouse pulmonary alveoli revealed by an in vivo cryotechnique in combination with freeze-substitution

2000 ◽  
Vol 197 (2) ◽  
pp. 199-205 ◽  
Author(s):  
ICHIRO TAKAYAMA ◽  
NOBUO TERADA ◽  
TAKESHI BABA ◽  
HIDEHO UEDA ◽  
YASUHISA FUJII ◽  
...  
Keyword(s):  
Freeze Substitution ◽  
In Vivo Cryotechnique ◽  
Pulmonary Alveoli

Immunohistochemical analysis of various serum proteins in living mouse thymus with “in vivo cryotechnique”

2012 ◽  
Vol 45 (3) ◽  
pp. 129-139 ◽  
Author(s):  
Yuqin Bai ◽  
Bao Wu ◽  
Nobuo Terada ◽  
Yurika Saitoh ◽  
Nobuhiko Ohno ◽  
...  
Keyword(s):  
Serum Proteins ◽  
Immunohistochemical Analysis ◽  
In Vivo Cryotechnique ◽  
Mouse Thymus ◽  
Living Mouse

scholarly journals Immunoreactivity of Glutamate in Mouse Retina Inner Segment of Photoreceptors With In Vivo Cryotechnique

2009 ◽  
Vol 57 (9) ◽  
pp. 883-888 ◽  
Author(s):  
Nobuo Terada ◽  
Nobuhiko Ohno ◽  
Sei Saitoh ◽  
Yurika Saitoh ◽  
Shinichi Ohno
Keyword(s):  
Ganglion Cell ◽  
Freeze Substitution ◽  
Controversial Issue ◽  
Mouse Retina ◽  
In Vivo Cryotechnique ◽  
Low Concentration ◽  
Cell Layers

The purpose of this study was to clarify a previously controversial issue concerning glutamate (Glu) immunoreactivity (IR) in the inner segment (IS) of photoreceptors by using in vivo cryotechnique (IVCT) followed by freeze substitution (FS), which enabled us to analyze the cells and tissues reflecting living states. Eyeballs from anesthetized mice were directly frozen using IVCT. The frozen tissues were processed for FS fixation in acetone containing chemical fixatives, and embedded in paraffin. Deparaffinized sections were immunostained with an anti-Glu antibody. The strongest Glu-IR was obtained in the specimens prepared by FS with paraformaldehyde or a low concentration of glutaraldehyde, whereas no Glu-IR was obtained without the chemical fixatives. The Glu was immunolocalized in the IS, outer and inner plexiform and ganglion cell layers. Thus, the immunolocalization of Glu in the IS was clearly demonstrated using IVCT.

Visualization of ATP with Luciferin-Luciferase Reaction in Mouse Skeletal Muscles Using an “In Vivo Cryotechnique”

2012 ◽  
Vol 18 (5) ◽  
pp. 1030-1036 ◽  
Author(s):  
N. Terada ◽  
Y. Saitoh ◽  
S. Saitoh ◽  
N. Ohno ◽  
K. Fujishita ◽  
...  
Keyword(s):  
Skeletal Muscles ◽  
Muscle Fibers ◽  
Freeze Substitution ◽  
Histological Structure ◽  
Type I ◽  
In Vivo Cryotechnique ◽  
Tissue Sections ◽  
Microscopic Evaluation

AbstractAdenosine triphosphate (ATP) is a well-known energy source for muscle contraction. In this study, to visualize localization of ATP, a luciferin-luciferase reaction (LLR) was performed in mouse skeletal muscle with an “in vivo cryotechnique” (IVCT). First, to confirm if ATP molecules could be trapped and detected after glutaraldehyde (GA) treatment, ATP was directly attached to glass slides with GA, and LLR was performed. The LLR was clearly detected as an intentional design of the ATP attachment. The intensity of the light unit by LLR was correlated with the concentration of the GA-treated ATP in vitro. Next, LLR was evaluated in mouse skeletal muscles with IVCT followed by freeze-substitution fixation (FS) in acetone-containing GA. In such tissue sections the histological structure was well maintained, and the intensity of LLR in areas between muscle fibers and connective tissues was different. Moreover, differences in LLR among muscle fibers were also detected. For the IVCT-FS tissue sections, diaminobenzidine (DAB) reactions were clearly detected in type I muscle fibers and erythrocytes in capillaries, which demonstrated flow shape. Thus, it became possible to perform microscopic evaluation of the numbers of ATP molecules in the mouse skeletal muscles with IVCT, which mostly reflect living states.

Immunohistochemical detection of soluble immunoglobulins in living mouse small intestines using an in vivo cryotechnique

2010 ◽  
Vol 361 (1-2) ◽  
pp. 64-74 ◽  
Author(s):  
Satoshi Shimo ◽  
Sei Saitoh ◽  
Nobuo Terada ◽  
Nobuhiko Ohno ◽  
Yurika Saitoh ◽  
...  
Keyword(s):  
Immunohistochemical Detection ◽  
In Vivo Cryotechnique ◽  
Living Mouse ◽  
Small Intestines

X-Ray Microanalysis of Human Erythrocytes Artificially Jetted Through Tubes At Different Pressures by “in vitro Cryotechnique”

2001 ◽  
Vol 7 (S2) ◽  
pp. 732-733
Author(s):  
Y. Fujii ◽  
N. Terada ◽  
T. Baba ◽  
H. Ueda ◽  
S. Ohno
Keyword(s):  
Human Erythrocyte ◽  
Morphological Changes ◽  
Freeze Substitution ◽  
Substitution Method ◽  
In Vivo Cryotechnique ◽  
Blood Capillaries ◽  
X Ray ◽  
Freeze Dried

It is well known that flowing erythrocytes in blood capillaries were morphologically changing in vivo, as observed by light microscopy. Recently, dynamic morphological changes of flowing mouse erythrocytes in large blood vessels and hepatic sinusoids were demonstrated by scanning (SEM) or transmission (TEM) electron microscopy with our “in vivo cryotechnique”. Moreover, human erythrocyte deformability was already studied under artificially jetting conditions at different pressures by using “in vivo cryotechnique”, followed by freeze-substitution method for SEM. in this study, we have analyzed elemental changes of each human erythrocyte at different jetting pressures by “in vivo cryotechnique” combined with SEM for X-ray microanalysis.Human blood was collected with heparin-coated syringes, divided into two groups and kept at 4°C and 36°C. They were directly jetted into isopentane-propane cryogen (-193°C) through tubes (21 gauge) at different pressures (0-220mmHg) (Fig.la). The frozen blood samples were freeze-dried (4-6×10-7torr,-95°C,24h) in Eiko FD-3AS apparatus (Eiko Engineering, Japan) (Fig. lb).

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