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In-Depth Imaging Using Two-Photon Excitation Microscopy

Published online by Cambridge University Press:  02 July 2020

H.C. Gerritsen ,
J.M. Vroom  and
C.J. de Grauw
H.C. Gerritsen
Affiliation:
Debye Institute, Utrecht University, P.O. Box 80.000, 3508, TA, Utrecht, Netherlands
J.M. Vroom
Affiliation:
Debye Institute, Utrecht University, P.O. Box 80.000, 3508, TA, Utrecht, Netherlands
C.J. de Grauw
Affiliation:
Debye Institute, Utrecht University, P.O. Box 80.000, 3508, TA, Utrecht, Netherlands
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Extract

Two-Photon Excitation (TPE) microscopy is a powerful tool for the imaging of biological specimen. The most important advantages compared with conventional confocal microscopy are: intrinsic optical sectioning, reduced phototoxcicity and photobleaching, and an increased penetration into the specimen. The increased penetration arises from the use of the longer wavelength excitation light, in the red or near infra-red.

Here, a quantitative in-depth TPE imaging study is presented on dental biofilm. The biofilm contains a mixed culture of a large number of oral bacteria with a typical size of ≈ 1 μm diameter. They exhibit a complex microstructure and heterogeneous physiological and biochemical properties. The high density of the biofilms limits the depth range over which confocal images can be acquired to 25-40 μm.

Using a two-photon excitation microscope fluorescence intensity images could be recorded over the whole thickness (100 μm) of the biofilm sample.

Type
New Developments in Multi-Photon Excitation Microscopy
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 420 - 421
Copyright
Copyright © Microscopy Society of America

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