Differential polarization laser scanning microscopy. Biological applications

G. Steinbach, F. Besson, I. Pomozi, G. Garab

Research output: Chapter in Book/Report/Conference proceedingConference contribution

4 Citations (Scopus)

Abstract

With the aid of a differential polarization (DP) apparatus, developed in our laboratory and attached to our laser scanning confocal microscope, we can measure the magnitude and spatial distribution of 8 different DP quantities: linear and circular dichroism (LD&CD), linear and circular anisotropy of the emission (R and CPL, confocal), fluorescence detected dichroisms (FDLD&FDCD, confocal), linear birefringence (LB), and the degree of polarization of fluorescence emission (P, confocal). The attachment uses high frequency modulation and subsequent demodulation, via lock-in amplifier, of the detected intensity values, and records and displays pixel-by-pixel the measured DP quantity. These microscopic DP data carry important physical information on the molecular architecture of anisotropically organized samples. Microscopic DP measurements are thought to be of particular importance in biology. In most biological samples anisotropy is difficult to determine with conventional, macroscopic DP measurements and microscopic variations are of special significance. In this paper, we describe the method of LB imaging. Using magnetically oriented isolated chloroplasts trapped in polyacrylamide gel, we demonstrate that LB can be determined with high sensitivity and good spatial resolution. Granal thylakoid membranes in edge-aligned orientation exhibited strong LB, with large variations in its sign and magnitude. In face-aligned position LB was considerably weaker, and tended to vanish when averaged for the whole image. The strong local variations are attributed to the inherent heterogeneity of the membranes, i.e. to their internal differentiation into multilamellar, stacked membranes (grana), and single thylakoids (stroma membranes). Further details and applications of our DP-LSM will be published elsewhere.

Original languageEnglish
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
EditorsW.C.W. Chan, K. Yu, U.J. Krull, R.I. Hornsey, B.C. Wilson
Volume5969
DOIs
Publication statusPublished - 2005
EventPhotonic Applications in Biosensing and Imaging - Toronto, ON, Canada
Duration: Sep 12 2005Sep 14 2005

Other

OtherPhotonic Applications in Biosensing and Imaging
CountryCanada
CityToronto, ON
Period9/12/059/14/05

Fingerprint

Microscopic examination
Polarization
Scanning
Birefringence
Lasers
Membranes
Anisotropy
Pixels
Fluorescence
Dichroism
Frequency modulation
Demodulation
Polyacrylates
Spatial distribution
Microscopes
Gels
Display devices
Imaging techniques

Keywords

  • Anisotropy
  • Confocal laser scanning microscopy
  • Differential polarization
  • Linear birefringence
  • Thylakoid membrane

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Steinbach, G., Besson, F., Pomozi, I., & Garab, G. (2005). Differential polarization laser scanning microscopy. Biological applications. In W. C. W. Chan, K. Yu, U. J. Krull, R. I. Hornsey, & B. C. Wilson (Eds.), Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 5969). [59692C] https://doi.org/10.1117/12.639362

Differential polarization laser scanning microscopy. Biological applications. / Steinbach, G.; Besson, F.; Pomozi, I.; Garab, G.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. ed. / W.C.W. Chan; K. Yu; U.J. Krull; R.I. Hornsey; B.C. Wilson. Vol. 5969 2005. 59692C.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Steinbach, G, Besson, F, Pomozi, I & Garab, G 2005, Differential polarization laser scanning microscopy. Biological applications. in WCW Chan, K Yu, UJ Krull, RI Hornsey & BC Wilson (eds), Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 5969, 59692C, Photonic Applications in Biosensing and Imaging, Toronto, ON, Canada, 9/12/05. https://doi.org/10.1117/12.639362
Steinbach G, Besson F, Pomozi I, Garab G. Differential polarization laser scanning microscopy. Biological applications. In Chan WCW, Yu K, Krull UJ, Hornsey RI, Wilson BC, editors, Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 5969. 2005. 59692C https://doi.org/10.1117/12.639362
Steinbach, G. ; Besson, F. ; Pomozi, I. ; Garab, G. / Differential polarization laser scanning microscopy. Biological applications. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. editor / W.C.W. Chan ; K. Yu ; U.J. Krull ; R.I. Hornsey ; B.C. Wilson. Vol. 5969 2005.
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