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Proceedings Paper

Influence of sample preparation and identification of subcelluar structures in quantitative holographic phase contrast microscopy
Author(s): Björn Kemper; Lisa Schmidt; Sabine Przibilla; Christina Rommel; Angelika Vollmer; Steffi Ketelhut; Jürgen Schnekenburger; Gert von Bally
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Paper Abstract

Digital holographic microscopy (DHM) provides label-free quantitative phase contrast with low demands on sample preparation. Nevertheless, for DHM measurements on fixed cells the mounting medium has to be considered while the phase contrast of living cells may be influenced by the used buffer solution. To quantify these effects, the maximum cell caused phase contrast and the visibility of the nucleoli were analyzed. A second aim of the study was to identify subcellular components in DHM phase contrast images. Therefore, comparative investigations using bright field imaging, DHM and fluorescence microscopy with 4',6- Diamidino-2-phenylindol (DAPI) staining were performed. DAPI-staining visualizes cell components containing DNA. The obtained results demonstrate exemplarily for two tumor cell lines that from DHM phase contrast images of fixed cells in phosphate buffer saline (PBS) cell thickness values are obtained which are comparable to living cells. Furthermore, it is shown that in many cases nucleus components can be identified only by DHM phase contrast.

Paper Details

Date Published: 14 May 2010
PDF: 8 pages
Proc. SPIE 7715, Biophotonics: Photonic Solutions for Better Health Care II, 771504 (14 May 2010); doi: 10.1117/12.854004
Show Author Affiliations
Björn Kemper, Westfälische Wilhelms-Univ. Münster (Germany)
Lisa Schmidt, Westfälische Wilhelms-Univ. Münster (Germany)
Sabine Przibilla, Westfälische Wilhelms-Univ. Münster (Germany)
Christina Rommel, Westfälische Wilhelms-Univ. Münster (Germany)
Angelika Vollmer, Westfälische Wilhelms-Univ. Münster (Germany)
Steffi Ketelhut, Westfälische Wilhelms-Univ. Münster (Germany)
Jürgen Schnekenburger, Westfälische Wilhelms-Univ. Münster (Germany)
Gert von Bally, Westfälische Wilhelms-Univ. Münster (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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