Mapping microscopic order in plant and mammalian cells and tissues

Novel differential polarization attachment for new generation confocal microscopes (DP-LSM)

G. Steinbach, K. Pawlak, I. Pomozi, E. A. Tóth, A. Molnár, J. Matkó, G. Garab

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Elucidation of the molecular architecture of complex, highly organized molecular macro-assemblies is an important, basic task for biology. Differential polarization (DP) measurements, such as linear (LD) and circular dichroism (CD) or the anisotropy of the fluorescence emission (r), which can be carried out in a dichrograph or spectrofluorimeter, respectively, carry unique, spatially averaged information about the molecular organization of the sample. For inhomogeneous samples-e.g. cells and tissues-measurements on macroscopic scale are not satisfactory, and in some cases not feasible, thus microscopic techniques must be applied. The microscopic DP-imaging technique, when based on confocal laser scanning microscope (LSM), allows the pixel by pixel mapping of anisotropy of a sample in 2D and 3D. The first DPLSM configuration, which, in fluorescence mode, allowed confocal imaging of different DP quantities in real-time, without interfering with the 'conventional' imaging, was built on a Zeiss LSM410. It was demonstrated to be capable of determining non-confocally the linear birefringence (LB) or LD of a sample and, confocally, its FDLD (fluorescence detected LD), the degree of polarization (P) and the anisotropy of the fluorescence emission (r), following polarized and non-polarized excitation, respectively (Steinbach et al 2009 Acta Histochem.111 316-25). This DPLSM configuration, however, cannot simply be adopted to new generation microscopes with considerably more compact structures. As shown here, for an Olympus FV500, we designed an easy-to-install DP attachment to determine LB, LD, FDLD and r, in new-generation confocal microscopes, which, in principle, can be complemented with a Pimaging unit, but specifically to the brand and type of LSM.

Original languageEnglish
Article number015005
JournalMethods and Applications in Fluorescence
Volume2
Issue number1
DOIs
Publication statusPublished - Mar 1 2014

Fingerprint

attachment
Microscopes
Fluorescence
microscopes
Cells
Polarization
Tissue
Scanning
fluorescence
scanning
Lasers
polarization
Anisotropy
lasers
Birefringence
Imaging techniques
anisotropy
birefringence
Pixels
pixels

Keywords

  • Confocal imaging
  • Fluorescence
  • Microscopy
  • Polarization
  • Structural order

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Instrumentation
  • Spectroscopy
  • Materials Science(all)

Cite this

Mapping microscopic order in plant and mammalian cells and tissues : Novel differential polarization attachment for new generation confocal microscopes (DP-LSM). / Steinbach, G.; Pawlak, K.; Pomozi, I.; Tóth, E. A.; Molnár, A.; Matkó, J.; Garab, G.

In: Methods and Applications in Fluorescence, Vol. 2, No. 1, 015005, 01.03.2014.

Research output: Contribution to journalArticle

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