Structured illumination microscopy of autofluorescent aggregations in human tissue

Gerrit Best; Roman Amberger; David Baddeley; Thomas Ach; Stefan Dithmar; Rainer Heintzmann; Christoph Cremer

doi:10.1016/j.micron.2010.06.016

Structured illumination microscopy of autofluorescent aggregations in human tissue

Micron. 2011 Jun;42(4):330-5. doi: 10.1016/j.micron.2010.06.016. Epub 2010 Sep 15.

Authors

Gerrit Best¹, Roman Amberger¹, David Baddeley², Thomas Ach³, Stefan Dithmar³, Rainer Heintzmann⁴, Christoph Cremer⁵

Affiliations

¹ Applied Optics and Information Processing, Kirchhoff-Institute for Physics, University of Heidelberg, Heidelberg, Germany.
² Applied Optics and Information Processing, Kirchhoff-Institute for Physics, University of Heidelberg, Heidelberg, Germany; Department of Physiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
³ Department of Ophthalmology, University Hospital Heidelberg, Heidelberg, Germany.
⁴ Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom; Institute of Photonic Technology, Jena, Germany.
⁵ Applied Optics and Information Processing, Kirchhoff-Institute for Physics, University of Heidelberg, Heidelberg, Germany; Institute for Pharmacy and Molecular Biotechnology, University of Heidelberg, Heidelberg, Germany; Institute for Molecular Biophysics, The Jackson Laboratory, Bar Harbor, Maine, USA.

PMID: 20926302
DOI: 10.1016/j.micron.2010.06.016

Abstract

Sections from human eye tissue were analyzed with Structured Illumination Microscopy (SIM) using a specially designed microscope setup. In this microscope the structured illumination was generated with a Twyman-Green Interferometer. This SIM technique allowed us to acquire light-optical images of autofluorophore distributions in the tissue with previously unmatched optical resolution. In this work the unique setup of the microscope made possible the application of SIM with three different excitation wavelengths (488, 568 and 647 nm), thus enabling us to gather spectral information about the autofluorescence signal.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Eye / cytology
Humans
Lighting / methods*
Microscopy, Fluorescence / methods*