高圧凍結技法とは、物理学的に2,100 Bars（大気圧の約2,000倍）の高圧下では氷晶形成が抑制される原理に基づいた新たな凍結技法で、氷晶による細胞形態の損傷がない良好な凍結（硝子様凍結）が得られる凍結面からの深度は200 µmに達し、金属圧着法に代表される従来の凍結技法をはるかに凌ぎます。

高圧凍結技法によるラット胃粘膜表層部電顕観察像（G：粘液ゲル層)

凍結面から深さ100 µm（上図のBox内）の形態も良好に保持されています

従来の化学固定法と高圧凍結技法の形態像を比較してご覧下さい

胃粘液・漿液分泌の組織化学的研究
ラット単離胃粘膜実験モデル研究
上皮細胞剥離・更新機構の超微形態研究
胃酸分泌壁細胞の特異な膜動態研究


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Sawaguchi, A., Aoyama, F., Ide S., Goto, Y., Suganuma, T.: A new device for high-pressure freezing of cultured cell monolayer using 10 µm-thin stainless discs as both culture plate and specimen carrier.
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Aoyama, F., Sawaguchi, A., Ide, S., Kitamura, K., Suganuma, T.: Exfoliation of gastric pit-parietal cells into the gastric lumen associated with a stimulation of isolated rat gastric mucosa in vitro: a morphological study by the application of cryotechniques.
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Sawaguchi, A., Aoyama, F., Ohashi, M., Ide S., Suganuma, T.: Ultrastructural transformation of gastric parietal cells reverting from the active to the resting state of acid secretion revealed in isolated rat gastric mucosa model processed by high-pressure freezing.
J Electron Microsc 55: 97-105, 2006
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Sawaguchi, A., Aoyama, F., Ide, S., Suganuma, T.: The cryofixation of isolated rat gastric mucosa provides new insights into the functional transformation of gastric parietal cells: an in vitro experimental model study.
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Sawaguchi, A., Ide, S., Suganuma, T.: Application of 10-µm thin stainless foil to a new assembly of the specimen carrier in high pressure freezing.
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Sawaguchi, A., McDonald, K.L., Forte, J.G.: High-pressure freezing of isolated gastric glands provides new insight into the fine structure and subcellular localization of H+/K+-ATPase in gastric parietal cells.
J Histochem Cytochem, 52: 77-86, 2004
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Sawaguchi, A., Yao, X., Forte, J.G., McDonald, K.L.: Direct attachment of cell suspensions to high pressure freezing specimen planchettes.
J Microsc 212: 13-20, 2003
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Sawaguchi, A., McDonald, K.L., Karvar, S., Forte, J.G.: A new approach for high-pressure freezing of primary culture cells: the fine structure and stimulation-associated transformation of cultured rabbit gastric parietal cells.
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Sawaguchi, A., Ishihara, K., Kawano, J., Oinuma, T., Hotta, K., Suganuma, T.: Fluid dynamics of the excretory flow of zymogenic and mucin contents in rat gastric gland processed by high pressure freezing/freeze substitution.
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Sawaguchi, A., Tojo, H., Kawano, J., Okamoto, M., Suganuma, T.: Immunocytochemical demonstration of the secretory dynamics of zymogenic contents in rat gastric gland processed by high-pressure freezing/freeze substitution, with special references to phospholipase A2 and phospholipase Cg1.
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Sawaguchi, A., Ide, S., Goto, Y., Kawano, J., Oinuma, T., Suganuma, T.: A simple contrast enhancement by potassium permanganate oxidation for Lowicryl K4M ultrathin sections prepared by high pressure freezing/freeze substitution.
J Microsc 201: 77-83, 2001
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Sawaguchi, A., Ide, S., Kawano, J., Nagaike, R., Oinuma, T., Tojo, H., Okamoto, M., Suganuma, T.: Reappraisal of potassium permanganate oxidation applied to Lowicryl K4M embedded tissues processed by high pressure freezing / freeze substitution, with special reference to differential staining of the zymogen granules of rat gastric chief cells.
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澤口　朗：超微形態と生体物質の保存に優れた加圧凍結技法
Plant Morphology 25: 7-10, 2013
doi.org/10.5685/plmorphol.25.7

澤口　朗、豊嶋　典世：高圧凍結技法による胃粘膜の超微形態研究
顕微鏡 45: 130-132, 2010
doi.org/10.11410/kenbikyo.45.2_130

澤口　朗、菅沼龍夫：高圧凍結技法の新展開─培養細胞への応用
顕微鏡 403: 204-206, 2005
doi.org/10.11410/kenbikyo2004.40.204