Fluorescence-detected linear dichroism imaging in a re-scan confocal microscope equipped with differential polarization attachment

Gabor Steinbach, David Nagy, Gábor Sipka, Erik Manders, G. Garab, L. Zimányi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Confocal laser scanning microscopy is probably the most widely used and one of the most powerful techniques in basic biology, medicine and material sciences that is employed to elucidate the architecture of complex cellular structures and molecular macro-assemblies. It has recently been shown that the information content, signal-to-noise ratio and resolution of such microscopes (LSMs) can be improved significantly by adding different attachments or modifying their design, while retaining their user-friendly features and relatively moderate costs. Differential polarization (DP) attachments, using high-frequency modulation/demodulation circuits, have made LSMs capable of high-precision 2D and 3D mapping of the anisotropy of microscopic samples—without interfering with their ‘conventional’ fluorescence or transmission imaging (Steinbach et al. in Methods Appl Fluoresc 2:015005, 2014). The resolution and the quality of fluorescence imaging have been enhanced in the recently constructed Re-scan confocal microscopy (RCM) (De Luca et al. in Biomed Opt Express 4:2644–2656, 2013). In this work, we developed the RCM technique further, by adding a DP-attachment modulating the exciting laser beam via a liquid crystal (LC) retarder synchronized with the data acquisition system; by this means, and with the aid of a software, fluorescence-detected linear dichroism (FDLD), characteristic of the anisotropic molecular organization of the sample, could be recorded in parallel with the confocal fluorescence imaging. For demonstration, we show FDLD images of a plant cell wall (Ginkgo biloba) stained with Etzold’s staining solution.

Original languageEnglish
JournalEuropean Biophysics Journal
DOIs
Publication statusPublished - Jan 1 2019

Fingerprint

Confocal Microscopy
Fluorescence
Optical Imaging
Ginkgo biloba
Liquid Crystals
Anisotropy
Plant Cells
Signal-To-Noise Ratio
Cellular Structures
Information Systems
Cell Wall
Lasers
Software
Medicine
Staining and Labeling
Costs and Cost Analysis

Keywords

  • Anisotropy
  • DP-LSM
  • Fluorescence-detected linear dichroism
  • RCM

ASJC Scopus subject areas

  • Biophysics

Cite this

Fluorescence-detected linear dichroism imaging in a re-scan confocal microscope equipped with differential polarization attachment. / Steinbach, Gabor; Nagy, David; Sipka, Gábor; Manders, Erik; Garab, G.; Zimányi, L.

In: European Biophysics Journal, 01.01.2019.

Research output: Contribution to journalArticle

@article{a88d8477b9584cf2839ed5aa70cb22a8,
title = "Fluorescence-detected linear dichroism imaging in a re-scan confocal microscope equipped with differential polarization attachment",
abstract = "Confocal laser scanning microscopy is probably the most widely used and one of the most powerful techniques in basic biology, medicine and material sciences that is employed to elucidate the architecture of complex cellular structures and molecular macro-assemblies. It has recently been shown that the information content, signal-to-noise ratio and resolution of such microscopes (LSMs) can be improved significantly by adding different attachments or modifying their design, while retaining their user-friendly features and relatively moderate costs. Differential polarization (DP) attachments, using high-frequency modulation/demodulation circuits, have made LSMs capable of high-precision 2D and 3D mapping of the anisotropy of microscopic samples—without interfering with their ‘conventional’ fluorescence or transmission imaging (Steinbach et al. in Methods Appl Fluoresc 2:015005, 2014). The resolution and the quality of fluorescence imaging have been enhanced in the recently constructed Re-scan confocal microscopy (RCM) (De Luca et al. in Biomed Opt Express 4:2644–2656, 2013). In this work, we developed the RCM technique further, by adding a DP-attachment modulating the exciting laser beam via a liquid crystal (LC) retarder synchronized with the data acquisition system; by this means, and with the aid of a software, fluorescence-detected linear dichroism (FDLD), characteristic of the anisotropic molecular organization of the sample, could be recorded in parallel with the confocal fluorescence imaging. For demonstration, we show FDLD images of a plant cell wall (Ginkgo biloba) stained with Etzold’s staining solution.",
keywords = "Anisotropy, DP-LSM, Fluorescence-detected linear dichroism, RCM",
author = "Gabor Steinbach and David Nagy and G{\'a}bor Sipka and Erik Manders and G. Garab and L. Zim{\'a}nyi",
year = "2019",
month = "1",
day = "1",
doi = "10.1007/s00249-019-01365-4",
language = "English",
journal = "European Biophysics Journal",
issn = "0175-7571",
publisher = "Springer Verlag",

}

TY - JOUR

T1 - Fluorescence-detected linear dichroism imaging in a re-scan confocal microscope equipped with differential polarization attachment

AU - Steinbach, Gabor

AU - Nagy, David

AU - Sipka, Gábor

AU - Manders, Erik

AU - Garab, G.

AU - Zimányi, L.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Confocal laser scanning microscopy is probably the most widely used and one of the most powerful techniques in basic biology, medicine and material sciences that is employed to elucidate the architecture of complex cellular structures and molecular macro-assemblies. It has recently been shown that the information content, signal-to-noise ratio and resolution of such microscopes (LSMs) can be improved significantly by adding different attachments or modifying their design, while retaining their user-friendly features and relatively moderate costs. Differential polarization (DP) attachments, using high-frequency modulation/demodulation circuits, have made LSMs capable of high-precision 2D and 3D mapping of the anisotropy of microscopic samples—without interfering with their ‘conventional’ fluorescence or transmission imaging (Steinbach et al. in Methods Appl Fluoresc 2:015005, 2014). The resolution and the quality of fluorescence imaging have been enhanced in the recently constructed Re-scan confocal microscopy (RCM) (De Luca et al. in Biomed Opt Express 4:2644–2656, 2013). In this work, we developed the RCM technique further, by adding a DP-attachment modulating the exciting laser beam via a liquid crystal (LC) retarder synchronized with the data acquisition system; by this means, and with the aid of a software, fluorescence-detected linear dichroism (FDLD), characteristic of the anisotropic molecular organization of the sample, could be recorded in parallel with the confocal fluorescence imaging. For demonstration, we show FDLD images of a plant cell wall (Ginkgo biloba) stained with Etzold’s staining solution.

AB - Confocal laser scanning microscopy is probably the most widely used and one of the most powerful techniques in basic biology, medicine and material sciences that is employed to elucidate the architecture of complex cellular structures and molecular macro-assemblies. It has recently been shown that the information content, signal-to-noise ratio and resolution of such microscopes (LSMs) can be improved significantly by adding different attachments or modifying their design, while retaining their user-friendly features and relatively moderate costs. Differential polarization (DP) attachments, using high-frequency modulation/demodulation circuits, have made LSMs capable of high-precision 2D and 3D mapping of the anisotropy of microscopic samples—without interfering with their ‘conventional’ fluorescence or transmission imaging (Steinbach et al. in Methods Appl Fluoresc 2:015005, 2014). The resolution and the quality of fluorescence imaging have been enhanced in the recently constructed Re-scan confocal microscopy (RCM) (De Luca et al. in Biomed Opt Express 4:2644–2656, 2013). In this work, we developed the RCM technique further, by adding a DP-attachment modulating the exciting laser beam via a liquid crystal (LC) retarder synchronized with the data acquisition system; by this means, and with the aid of a software, fluorescence-detected linear dichroism (FDLD), characteristic of the anisotropic molecular organization of the sample, could be recorded in parallel with the confocal fluorescence imaging. For demonstration, we show FDLD images of a plant cell wall (Ginkgo biloba) stained with Etzold’s staining solution.

KW - Anisotropy

KW - DP-LSM

KW - Fluorescence-detected linear dichroism

KW - RCM

UR - http://www.scopus.com/inward/record.url?scp=85064478959&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85064478959&partnerID=8YFLogxK

U2 - 10.1007/s00249-019-01365-4

DO - 10.1007/s00249-019-01365-4

M3 - Article

AN - SCOPUS:85064478959

JO - European Biophysics Journal

JF - European Biophysics Journal

SN - 0175-7571

ER -