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A new HPF specimen carrier adapter for the use of high-pressure freezing with cryo-scanning electron microscope: two applications: stearic acid organization in a hydroxypropyl methylcellulose matrix and mice myocardium .

Payre, Bruno and Gontier, Etienne and Jarray, Ahmed and Martinez, Yves and Laugier, Jean Pierre and Delalleau, Alexandre and Gaillard-Martinie, Brigitte and Anselme, Isabelle and Goudouneche, Dominique and Fourquaux, Isabelle and Hemati, Mehrdji and Gerbaud, Vincent and Delisle, Marie Bernadette and Guilbeau-Frugier, Céline A new HPF specimen carrier adapter for the use of high-pressure freezing with cryo-scanning electron microscope: two applications: stearic acid organization in a hydroxypropyl methylcellulose matrix and mice myocardium . (2018) Journal of Microscopy, 271 (3). 255-265. ISSN 0022-2720

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Official URL: https://doi.org/10.1111/jmi.12713

Abstract

Cryogenic transmission electron microscopy of high-pressure freezing (HPF) samples is a well-established technique for the analysis of liquid containing specimens. This technique enables observation without removing water or other volatile components. The HPF technique is less used in scanning electron microscopy (SEM) due to the lack of a suitable HPF specimen carrier adapter. The traditional SEM cryotransfer system (PP3000T Quorum Laughton, East Sussex, UK; Alto Gatan, Pleasanton, CA, USA) usually uses nitrogen slush. Unfortunately, and unlike HPF, nitrogen slush produces water crystal artefacts. So, we propose a new HPF specimen carrier adapter for sample transfer from HPF system to cryogenic-scanning electronic microscope (Cryo-SEM). The new transfer system is validated using technical two applications, a stearic acid in hydroxypropyl methylcellulose solution and mice myocardium. Preservation of samples is suitable in both cases. Cryo-SEM examination of HPF samples enables a good correlation between acid stearic liquid concentration and acid stearic occupation surface (only for homogeneous solution). For biological samples as myocardium, cytoplasmic structures of cardiomyocyte are easily recognized with adequate preservation of organelle contacts and inner cell organization. We expect this new HPF specimen carrier adapter would enable more SEM-studies using HPF.

Item Type:Article
Additional Information:in presse 14 juin 2018 "first published" date sur le site 27 avril 2018 sur le document pdf.
HAL Id:hal-01831910
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Centre National de la Recherche Scientifique - CNRS (FRANCE)
Université de Toulouse > Institut National Polytechnique de Toulouse - INPT (FRANCE)
Université de Toulouse > Université Toulouse III - Paul Sabatier - UPS (FRANCE)
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Deposited By: Vincent GERBAUD
Deposited On:06 Jul 2018 10:49

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