Abstract Detail

Development and Structure

Pegg, Timothy [1], Gladish, Daniel [2], Baker, Robert (Rob) [3].

Autofluorescence for rapid visualization of plant anatomy among diverse taxa.

Fluorescence microscopy is an effective tool for viewing plant internal anatomy. However, using fluorescent antibodies or labels hinders throughput. We present a minimal protocol that takes advantage of inherent autofluorescence and aldehyde-induced fluorescence in plant cellular and sub-cellular structures to markedly increased throughput in cellular and ultrastructural visualization. Eleven species distributed across the plant phylogeny were subjected five fixatives. Samples prepared by embedding and mechanically sectioning or via whole mount. A confocal laser scanning system was used to collected micrographs. We evaluated and compared fixative influence on structural preservation and tissue autofluorescence. Formaldehyde-fixation of Viridaeplantae taxa samples generates useful structural data while requiring no additional histological staining or clearing. In addition, image acquisition requires only minimal specialized equipment in the form of fluorescence-capable microscopes. The minimal protocol developed in this experiment enables high throughput sample processing by eliminating the need for multi-day preparations.

1 - Miami University, 330 West Withrow Street, Oxford, OH, 45056, United States
2 - Miami University, Department Of Biology, 1601 University Blvd., Hamilton, OH, 45011, United States
3 - Miami University, Biology Department, 700 E High St, Pearson 390, Oxford, OH, 45056, United States

Keywords:
Viridaeplantae
anatomy
aldehyde
cellular
fixation
methods
Laser Scanning Confocal Microscopy
sub-cellular
throughput
autofluorescence.

Presentation Type: Oral Paper
Session: DS3, Development and Structure III
Location: /
Date: Thursday, July 22nd, 2021
Time: 10:45 AM(EDT)
Number: DS3004
Abstract ID:594
Candidate for Awards:Katherine Esau Award,Developmental and Structural Section Graduate Student Registration Award