4.3 Article Proceedings Paper

A rapid microbiopsy system to improve the preservation of biological samples prior to high-pressure freezing

Journal

JOURNAL OF MICROSCOPY
Volume 212, Issue -, Pages 3-12

Publisher

WILEY-BLACKWELL
DOI: 10.1046/j.1365-2818.2003.01226.x

Keywords

cryofixation; cryosectioning; electron microscopy; freeze-substitution; high-pressure freezing; microbiopsy

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A microbiopsy system for fast excision and transfer of biological specimens from donor to high-pressure freezer was developed. With a modified, commercially available, Promag 1.2 biopsy gun, tissue samples can be excised with a size small enough (0.6 mm x 1.2 mm x 0.3 mm) to be easily transferred into a newly designed specimen platelet. A self-made transfer unit allows fast transfer of the specimen from the needle into the specimen platelet. The platelet is then fixed in a commercially available specimen holder of a high-pressure freezing machine (EM PACT, Leica Microsystems, Vienna, Austria) and frozen therein. The time required by a well-instructed (but not experienced) person to execute all steps is in the range of half a minute. This period is considered short enough to maintain the excised tissue pieces close to their native state. We show that a range of animal tissues (liver, brain, kidney and muscle) are well preserved. To prove the quality of freezing achieved with the system, we show vitrified ivy leaves high-pressure frozen in the new specimen platelet.

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