title: Renaissance of morphological studies: the examination of functional structures in living animal organs using the in vivo cryotechnique.
abstract: Medical and biological scientists wish to understand the in vivo structures of the cells and tissues that make up living animal organs, as well as the locations of their molecular components. Recently, the live imaging of animal cells and tissues with fluorescence-labeled proteins produced via gene manipulation has become increasingly common. Therefore, it is important to ensure that findings derived from histological or immunohistochemical tissue sections of living animal organs are compatible with those obtained from live images of the same organs, which can be assessed using recently developed digital imaging techniques. Over the past two decades, we have performed immunohistochemical and morphological studies of the cells and tissues in living animal organs using a novel in vivo cryotechnique. The use of a specially designed liquid cryogen system with or without a cryoknife during this cryotechnique solved the technical problems that inevitably arise during the conventional preparation methods employed prior to light or electron microscopic examinations. Our in vivo cryotechnique has been found to be extremely useful for arresting transient physiological processes in cells and tissues and for maintaining their functional components-such as rapidly changing signaling molecules, membrane channels, or receptors-in situ. The purpose of the present review is to describe the basic mechanism underlying cryotechniques and the significance of our in vivo cryotechnique. In addition, it describes various morphological or immunohistochemical findings, observations made using quantum dots, and a Raman cryomicroscopy-based method for assessing oxygen saturation in the erythrocytes flowing through intestinal tissues.
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo, Japan. sohno@yamanashi.ac.jp.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Ohno
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo, Japan.
authors.forename: Yurika
authors.initials: Y
authors.lastname: Saitoh
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo, Japan.|Division of Neurobiology and Bioinformatics, National Institute for Physiological Sciences, Okazaki, Japan.
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Ohno
authors.affiliation: Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo, Japan.|Division of Health Sciences, Shinshu University Graduate School of Medicine, Matsumoto, Japan.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Terada
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003952:Diagnostic Imaging / Q000379:methods*; D004912:Erythrocytes / Q000378:metabolism / Q000502:physiology; D006439:Hemodynamics; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007668:Kidney / Q000000981:diagnostic imaging* / Q000648:ultrastructure; D008168:Lung / Q000000981:diagnostic imaging* / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008853:Microscopy; D009132:Muscles / Q000000981:diagnostic imaging* / Q000648:ultrastructure; D010100:Oxygen / Q000097:blood; D010420:Pentanes; D011407:Propane; D045663:Quantum Dots; D016707:Tissue Fixation / Q000379:methods
chemical_list: D010420:Pentanes; D010100:Oxygen; D011407:Propane; C067038:isopentane
keywords: Bioimaging; Functional morphology; In vivo cryotechnique; Living animal organ; Soluble component