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

B. Payre*, E. Gontier, A. Jarray, Y. Martinez, J.P. Laugier, A. Delalleau, B.M. Gaillard, I. Anselme, D. Goudouneche, I. Fourquaux, M. Hemati, V. Gerbaud, M.B. Delisle, C. Guilbeau-Frugier

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

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    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
    Original languageEnglish
    Pages (from-to)255-265
    Number of pages11
    JournalJournal of microscopy
    Volume271
    Issue number3
    DOIs
    Publication statusPublished - Sep 2018

    Keywords

    • Cryogenic scanning electron microscopy
    • Cryogenic fracture
    • Cryogenic freezing
    • High-pressure freezing
    • HPF specimen carrier adapter
    • HPMC
    • Stearic acid
    • SEM

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