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Proceedings Paper

Phase resolved and coherence gated en face reflection imaging of multilayered embryonal carcinoma cells
Author(s): Toyohiko Yamauchi; Tadashi Fukami; Hidenao Iwai; Yutaka Yamashita
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Paper Abstract

Embryonal carcinoma (EC) cells, which are cell lines derived from teratocarcinomas, have characteristics in common with stem cells and differentiate into many kinds of functional cells. Similar to embryonic stem (ES) cells, undifferentiated EC cells form multi-layered spheroids. In order to visualize the three-dimensional structure of multilayered EC cells without labeling, we employed full-field interference microscopy with the aid of a low-coherence quantitative phase microscope, which is a reflection-type interference microscope employing the digital holographic technique with a low-coherent light source. Owing to the low-coherency of the light-source (halogen lamp), only the light reflected from reflective surface at a specific sectioning height generates an interference image on the CCD camera. P19CL6 EC cells, derived from mouse teratocarcinomas, formed spheroids that are about 50 to 200 micrometers in diameter. Since the height of each cell is around 10 micrometers, it is assumed that each spheroid has 5 to 20 cell layers. The P19CL6 spheroids were imaged in an upright configuration and the horizontally sectioned reflection images of the sample were obtained by sequentially and vertically scanning the zero-path-length height. Our results show the threedimensional structure of the spheroids, in which plasma and nuclear membranes were distinguishably imaged. The results imply that our technique is further capable of imaging induced pluripotent stem (iPS) cells for the assessment of cell properties including their pluripotency.

Paper Details

Date Published: 9 February 2012
PDF: 10 pages
Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250G (9 February 2012); doi: 10.1117/12.907637
Show Author Affiliations
Toyohiko Yamauchi, Hamamatsu Photonics K.K. (Japan)
Tadashi Fukami, Hamamatsu Photonics K.K. (Japan)
Hidenao Iwai, Hamamatsu Photonics K.K. (Japan)
Yutaka Yamashita, Hamamatsu Photonics K.K. (Japan)

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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