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Proceedings Paper

SIM and PALM: high-resolution microscopy methods and their consequences for cell biology
Author(s): Gerhard Krampert; Ingo Kleppe; Thomas Kalkbrenner; Klaus Weisshart; Ralf Wolleschensky; Michael Kempe
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Paper Abstract

The diffraction limit in traditional fluorescence microscopy (approximately 200 and 600 nanometers in lateral and axial directions, respectively) has restricted the applications in bio-medical research. However, over the last 10 years various techniques have emerged to overcome this limit. Each of these techniques has its own characteristics that influence its application in biology. This paper will show how two of the techniques, Structured Illumination Microscopy (SIM) and PhotoActivated Localization Microscopy (PALM), complement each other in imaging of biological samples beyond the resolution of classical widefield fluorescence microscopy. As a reference the properties of two well known standard imaging techniques in this field, confocal Laser Scanning Microscopy (LSM) and Total Internal Reflection (TIRF) microscopy, are compared to the properties of the two high resolution techniques. Combined SIM/PALM imaging allows the extremely accurate localization of individual molecules within the context of various fluorescent structures already resolved in 3D with a resolution of up to 100nm using SIM. Such a combined system provides the biologist with an unprecedented view of the sub-cellular organization of life.

Paper Details

Date Published: 14 May 2010
PDF: 9 pages
Proc. SPIE 7715, Biophotonics: Photonic Solutions for Better Health Care II, 771503 (14 May 2010); doi: 10.1117/12.854633
Show Author Affiliations
Gerhard Krampert, Carl Zeiss AG (Germany)
Ingo Kleppe, Carl Zeiss MicroImaging GmbH (Germany)
Thomas Kalkbrenner, Carl Zeiss MicroImaging GmbH (Germany)
Klaus Weisshart, Carl Zeiss MicroImaging GmbH (Germany)
Ralf Wolleschensky, Carl Zeiss MicroImaging GmbH (Germany)
Michael Kempe, Carl Zeiss AG (Germany)

Published in SPIE Proceedings Vol. 7715:
Biophotonics: Photonic Solutions for Better Health Care II
Jürgen Popp; Wolfgang Drexler; Valery V. Tuchin; Dennis L. Matthews, Editor(s)

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