Imaging anisotropy using differential polarization laser scanning confocal microscopy

Gábor Steinbach, István Pomozi, Ottó Zsiros, László Menczel, Gyozo Garab

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

We have constructed differential polarization (DP) attachments to a laser scanning microscope (LSM) for imaging the main DP quantities of anisotropic microscopic objects. The DP-LSM operates with high-frequency modulation and subsequent demodulation and displays the main DP quantities pixel by pixel. These, for linearly polarized light, include: (i) linear birefringence (LB), which is exhibited by structurally and/or optically anisotropic material; (ii) linear dichroism (LD), which carries information on the anisotropic distribution of the molecules, i.e. of their absorbance transition dipole vectors, in the sample; (iii) fluorescence-detected LD (FDLD), which carries the same information for fluorescent dyes upon excitations with two orthogonally polarized light beams; (iv) anisotropy of the fluorescence emission (r), excited with non-polarized light, which is determined by the distribution of the emission transition dipole vectors in the sample and is analogous with LD and (v) the degree of polarization of the fluorescence emission (P), excited with polarized light, which depends on the depolarization of the emission e.g. due to the rotation of molecules during their excitation lifetimes. In fluorescence regimes, the DP images can be recorded in the confocal regime of the microscope, which thus warrants good spatial resolution and the possibility of mapping the anisotropy in three dimensions. In this paper, we outline the design and technical realization of our DP-LSM and give a few examples on DP imaging of different biological samples.

Original languageEnglish
Pages (from-to)317-326
Number of pages10
JournalActa histochemica
Volume111
Issue number4
DOIs
Publication statusPublished - Jul 2009

Keywords

  • Anisotropy
  • Birefringence
  • Confocal laser scanning microscopy
  • Fluorescence-detected linear dichroism
  • Linear dichroism
  • Polarization of the fluorescence emission

ASJC Scopus subject areas

  • Histology
  • Cell Biology

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