Imaging fluorescence detected linear dichroism of plant cell walls in laser scanning confocal microscope

Gábor Steinbach, István Pomozi, Ottó Zsiros, Anikó Páy, Gábor V. Horváth, Gyozo Garab

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Anisotropy carries important information on the molecular organization of biological samples. Its determination requires a combination of microscopy and polarization spectroscopy tools. The authors constructed differential polarization (DP) attachments to a laser scanning microscope in order to determine physical quantities related to the anisotropic distribution of molecules in microscopic samples; here the authors focus on fluorescence- detected linear dichroism (FDLD). By modulating the linear polarization of the laser beam between two orthogonally polarized states and by using a demodulation circuit, the authors determine the associated transmitted and fluorescence intensity-difference signals, which serve the basis for LD (linear dichroism) and FDLD, respectively. The authors demonstrate on sections of Convallaria majalis root tissue stained with Acridin Orange that while (nonconfocal) LD images remain smeared and weak, FDLD images recorded in confocal mode reveal strong anisotropy of the cell wall. FDLD imaging is suitable for mapping the anisotropic distribution of transition dipoles in 3 dimensions. A mathematical model is proposed to account for the fiber-laminate ultrastructure of the cell wall and for the intercalation of the dye molecules in complex, highly anisotropic architecture.

Original languageEnglish
Pages (from-to)202-208
Number of pages7
JournalCytometry Part A
Volume73
Issue number3
DOIs
Publication statusPublished - Mar 1 2008

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Keywords

  • Anisotropy
  • Cell wall
  • Cellulose
  • Confocal laser scanning microscopy
  • Fluorescence detected linear dichroism
  • Linear dichroism
  • Polarized light

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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