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Proceedings Paper

Two-photon cryomicroscope
Author(s): H. G. Breunig; C. Köhler; K. König
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Paper Abstract

We report on a new two-photon cryomicroscope which consist of a compact laser-scanning microscope combined with a motorized heating and freezing stage. Samples can be cooled down to -196 °C (77 K) and heated up to 600 °C (873 K) with adjustable heating/freezing rates between 0.01 K / min and 150 K / min. Two-photon imaging is realized by near infrared femtosecond-laser pulse excitation. The abilities of the two-photon cryomicroscope are illustrated in several measurements: imaging of fluorescent microspheres inside a piece of ice illustrates the feasibility of deep-microscopic imaging inside frozen sample. The temperature-dependent structural integrity of collagen is monitored by detection of second harmonic generation signals from porcine cornea. The measurements reveal also the dependence of the collagendenaturation temperature on hydration state of the cornea collagen. Furthermore, the potential of the two-photon cryomicroscope for optimization of freezing and thawing procedures as well as to evaluate the viability of frozen cells and tissue is discussed.

Paper Details

Date Published: 9 February 2012
PDF: 7 pages
Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 822516 (9 February 2012); doi: 10.1117/12.908348
Show Author Affiliations
H. G. Breunig, JenLab GmbH (Germany)
C. Köhler, JenLab GmbH (Germany)
K. König, JenLab GmbH (Germany)
Saarland Univ. (Germany)

Published in SPIE Proceedings Vol. 8225:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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