High resolution detection of uncoated metaphase chromosomes by means of field emission scanning electron microscopy

R. Rizzoli, E. Rizzi, M. Falconi, A. Galanzi, B. Baratta, G. Lattanzi, M. Vitale, L. Manzoli, G. Mazzotti

Research output: Contribution to journalArticlepeer-review

Abstract

HeLa metaphase chromosomes were exammed by means of "in lens" field emission scanning electron microscopy, which permits high resolution detection of uncoated biological samples. By using uncoated chromosomes as a model for comparison we report evidence of how traditional scanning electron microscopy techniques such as metal coating and conductive methods can generate errors in chromosome structure evaluation, since both give rise to morphological artifacts. By comparing the morphology of uncoated chromosomes obtained by two different isolation procedures, such as that utilized in standard cytogenetics and the polyamine method, we have drawn the following conclusions: (a) the standard cytogenetic method gives rise to a chromosome structure consisting of a flattened network of 10 nm fibers, in which higher order chromatin organization is absent. (b) Chromosomes obtained by the polyamine method show both three-dimensional profile and higher level folding of chromatin fibers, supporting the loop chromosome organization previously suggested by scanning electron microscopy observation of hexylene glycol isolated chromosomes.

Original languageEnglish
Pages (from-to)393-400
Number of pages8
JournalChromosoma
Volume103
Issue number6
DOIs
Publication statusPublished - Oct 1994

ASJC Scopus subject areas

  • Genetics

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