Routine fluorescence microscopy of single untethered protein molecules confined to a planar zone

Hongwei Gai, Gary A. Griess, Borries Demeler, Susan T. Weintraub, Philip Serwer

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

To bypass limitations of ensemble averaging biochemical analysis, microscopy-based detection and tracking are needed for single protein molecules that are diffusing in aqueous solution. Confining the molecules to a planar zone dramatically assists tracking. Procedures of microscopy should be routine enough so that effort is focused on the biochemistry. Fluorescence microscopy and partial planar confinement of single, untethered, aqueous protein molecules have been achieved here by use of a routine procedure. With this procedure, multiple thermally diffusing Alexa 488-stained bovine serum albumin molecules were observed during partial confinement to a thin aqueous zone next to a cover slip. The procedure produces confinement by partial re-swelling of a previously dried agarose gel on the microscope slide. Confinement was confirmed through analysis that revealed thermal motion lower in the third dimension than it was in the plane of observation.

Original languageEnglish (US)
Pages (from-to)256-262
Number of pages7
JournalJournal of Microscopy
Volume226
Issue number3
DOIs
StatePublished - Jun 2007

Keywords

  • Agarose
  • Alexa dye
  • Bovine serum albumin
  • Restricted thermal motion
  • Single-molecule biochemistry

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

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