A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells

Abstract The scanning electron microscope (SEM) has been reassembled into a new type of cryo-electron microscope (cryo-TSEM) by installing a new cryo-transfer holder and anti-contamination trap, which allowed simultaneous acquisition of both transmission images (STEM images) and surface images (SEM...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jiro Usukura, Akihiro Narita, Tomoharu Matsumoto, Eiji Usukura, Takeshi Sunaoshi, Syunya Watanabe, Yusuke Tamba, Yasuhira Nagakubo, Takashi Mizuo, Junzo Azuma, Masako Osumi, Kazutaka Nimura, Ryuichiro Tamochi, Yoichi Ose
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b8e0ce76230b4113a0fa5d7ebebda2c7
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:b8e0ce76230b4113a0fa5d7ebebda2c7
record_format	dspace
spelling	oai:doaj.org-article:b8e0ce76230b4113a0fa5d7ebebda2c72021-11-08T10:49:00ZA cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells10.1038/s41598-021-00979-z2045-2322https://doaj.org/article/b8e0ce76230b4113a0fa5d7ebebda2c72021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-00979-zhttps://doaj.org/toc/2045-2322Abstract The scanning electron microscope (SEM) has been reassembled into a new type of cryo-electron microscope (cryo-TSEM) by installing a new cryo-transfer holder and anti-contamination trap, which allowed simultaneous acquisition of both transmission images (STEM images) and surface images (SEM images) in the frozen state. The ultimate temperatures of the holder and the trap reached − 190 °C and − 210 °C, respectively, by applying a liquid nitrogen slush. The STEM images at 30 kV were comparable to, or superior to, the images acquired with conventional transmission electron microscope (100 kV TEM) in contrast and sharpness. The unroofing method was used to observe membrane cytoskeletons instead of the frozen section and the FIB methods. Deep sublimation of ice surrounding unroofed cells by regulating temperature enabled to emerge intracellular fine structures in thick frozen cells. Hence, fine structures in the vicinity of the cell membrane such as the cytoskeleton, polyribosome chains and endoplasmic reticulum (ER) became visible. The ER was distributed as a wide, flat structure beneath the cell membrane, forming a large spatial network with tubular ER.Jiro UsukuraAkihiro NaritaTomoharu MatsumotoEiji UsukuraTakeshi SunaoshiSyunya WatanabeYusuke TambaYasuhira NagakuboTakashi MizuoJunzo AzumaMasako OsumiKazutaka NimuraRyuichiro TamochiYoichi OseNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jiro Usukura Akihiro Narita Tomoharu Matsumoto Eiji Usukura Takeshi Sunaoshi Syunya Watanabe Yusuke Tamba Yasuhira Nagakubo Takashi Mizuo Junzo Azuma Masako Osumi Kazutaka Nimura Ryuichiro Tamochi Yoichi Ose A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells
description	Abstract The scanning electron microscope (SEM) has been reassembled into a new type of cryo-electron microscope (cryo-TSEM) by installing a new cryo-transfer holder and anti-contamination trap, which allowed simultaneous acquisition of both transmission images (STEM images) and surface images (SEM images) in the frozen state. The ultimate temperatures of the holder and the trap reached − 190 °C and − 210 °C, respectively, by applying a liquid nitrogen slush. The STEM images at 30 kV were comparable to, or superior to, the images acquired with conventional transmission electron microscope (100 kV TEM) in contrast and sharpness. The unroofing method was used to observe membrane cytoskeletons instead of the frozen section and the FIB methods. Deep sublimation of ice surrounding unroofed cells by regulating temperature enabled to emerge intracellular fine structures in thick frozen cells. Hence, fine structures in the vicinity of the cell membrane such as the cytoskeleton, polyribosome chains and endoplasmic reticulum (ER) became visible. The ER was distributed as a wide, flat structure beneath the cell membrane, forming a large spatial network with tubular ER.
format	article
author	Jiro Usukura Akihiro Narita Tomoharu Matsumoto Eiji Usukura Takeshi Sunaoshi Syunya Watanabe Yusuke Tamba Yasuhira Nagakubo Takashi Mizuo Junzo Azuma Masako Osumi Kazutaka Nimura Ryuichiro Tamochi Yoichi Ose
author_facet	Jiro Usukura Akihiro Narita Tomoharu Matsumoto Eiji Usukura Takeshi Sunaoshi Syunya Watanabe Yusuke Tamba Yasuhira Nagakubo Takashi Mizuo Junzo Azuma Masako Osumi Kazutaka Nimura Ryuichiro Tamochi Yoichi Ose
author_sort	Jiro Usukura
title	A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells
title_short	A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells
title_full	A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells
title_fullStr	A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells
title_full_unstemmed	A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells
title_sort	cryo-tsem with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b8e0ce76230b4113a0fa5d7ebebda2c7
work_keys_str_mv	AT jirousukura acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT akihironarita acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT tomoharumatsumoto acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT eijiusukura acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT takeshisunaoshi acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT syunyawatanabe acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT yusuketamba acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT yasuhiranagakubo acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT takashimizuo acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT junzoazuma acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT masakoosumi acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT kazutakanimura acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT ryuichirotamochi acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT yoichiose acryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT jirousukura cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT akihironarita cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT tomoharumatsumoto cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT eijiusukura cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT takeshisunaoshi cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT syunyawatanabe cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT yusuketamba cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT yasuhiranagakubo cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT takashimizuo cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT junzoazuma cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT masakoosumi cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT kazutakanimura cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT ryuichirotamochi cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells AT yoichiose cryotsemwithtemperaturecyclingcapabilityallowsdeepsublimationoficetouncoverfinestructuresinthickcells
_version_	1718442656188596224

A cryo-TSEM with temperature cycling capability allows deep sublimation of ice to uncover fine structures in thick cells

Ejemplares similares