Ultrastructural Localization of Hair Keratins in Human Scalp Hair Shafts as Revealed by Rapid-Freezing Immunocytochemistry.

Takami, Takeshi; Tajima, Toshihiro; Uchiwa, Hideyo; Arai, Seiichi

doi:10.1267/ahc.31.237

Cited by 3 publications

(2 citation statements)

References 29 publications

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“…25,34,68,127 For immunoelectron microscopy, follicular sections were first labelled with primary and secondary antibodies. 62,113,128 If silver enhancement was required, grids were poststained immediately following this procedure. 61 Recently, it has been established that dark-field scanning TEM (DF-STEM) produces sufficient contrast to detect ultra-small gold labels, about 5 nm gold, in an unstained section without requiring silver enhancement procedures.…”

Section: P Os T-s Tainingmentioning

confidence: 99%

“…However, the additional level of ultrastructural detail acquired following post‐staining with uranyl acetate and lead citrate was remarkable 25,34,68,127 . For immunoelectron microscopy, follicular sections were first labelled with primary and secondary antibodies 62,113,128 . If silver enhancement was required, grids were post‐stained immediately following this procedure 61 .…”

Section: Post‐stainingmentioning

confidence: 99%

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Processing hair follicles for transmission electron microscopy

Velamoor

Mitchell

Bostina

et al. 2021

Experimental Dermatology

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Transmission electron microscopy (TEM) has greatly advanced our knowledge of hair growth and follicle morphogenesis, but complex preparations such as fixation, dehydration and embedding compromise ultrastructure. While recent developments with cryofixation have been shown to preserve the ultrastructure of biological materials close to native state, they do have limitations. This review will focus on each stage of the TEM sample preparation process and their effects on the structural integrity of follicles.

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Section: P Os T-s Tainingmentioning

confidence: 99%

Section: Post‐stainingmentioning

confidence: 99%

Processing hair follicles for transmission electron microscopy

Velamoor

Mitchell

Bostina

et al. 2021

Experimental Dermatology

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Immuno-electron Microscopic Localization of Endothelial Nitric Oxide Synthase in Human Placental Terminal Villous Trophoblasts—Normal and Pre-eclamptic Pregnancy

et al. 2001

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Ultrastructural Localization of S100A3, a Cysteine-rich, Calcium Binding Protein, in Human Scalp Hair Shafts Revealed by Rapid-freezing Immunocytochemistry

Takami

Tajima

Arai³

et al. 1999

J Histochem Cytochem.

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SUMMARYWe have characterized the subcellular distribution of S100A3, a cysteine-rich calcium binding protein, in human scalp hair shaft. This was accomplished using rapidfreezing immunocytochemistry, a technique that combines rapid-freezing, freeze-substitution fixation without chemical fixatives, and subsequent electron microscopic detection of immunocytochemical labeling. This technique preserves both the antigenicity and the ultrastructural integrity of fully keratinized tissues, which are highly unmanageable when prepared for immunoelectron microscopy. In the hair shaft, S100A3 was primarily identified in the endocuticle and was also present in the intermacrofibrillar matrix surrounding macrofibril bundles of intermediate filament keratins in cortex cells. Double immunolabeling of S100A3 and hair keratins revealed the in situ spatial relationship between them. In the endocuticle, S100A3 was present on the inner portion of the endocuticle adjacent to the cell membrane complex, whereas hair keratins were present on the outer portion. These results provide the first ultrastructural evidence that an S100 protein is localized in specific subcompartments in human hair cells.

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Ultrastructural Localization of Hair Keratins in Human Scalp Hair Shafts as Revealed by Rapid-Freezing Immunocytochemistry.

Cited by 3 publications

References 29 publications

Processing hair follicles for transmission electron microscopy

Processing hair follicles for transmission electron microscopy

Immuno-electron Microscopic Localization of Endothelial Nitric Oxide Synthase in Human Placental Terminal Villous Trophoblasts—Normal and Pre-eclamptic Pregnancy

Ultrastructural Localization of S100A3, a Cysteine-rich, Calcium Binding Protein, in Human Scalp Hair Shafts Revealed by Rapid-freezing Immunocytochemistry

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