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

J Microsc. 2003 Oct;212(Pt 1):3-12. doi: 10.1046/j.1365-2818.2003.01226.x.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biopsy
  • Brain / ultrastructure
  • Cryopreservation / instrumentation*
  • Cryopreservation / methods*
  • Cryoultramicrotomy
  • Freeze Substitution
  • Freezing
  • Kidney / ultrastructure
  • Liver / ultrastructure
  • Microscopy, Electron
  • Muscles / ultrastructure
  • Plant Leaves / ultrastructure
  • Pressure
  • Rats
  • Specimen Handling / methods
  • Time Factors