Measurement of topographic phase image of living cells by white-light phase-shifting microscope with active stabilization of optical path difference

Toyohiko Yamauchi; Hidenao Iwai; Mitsuharu Miwa; Yutaka Yamashita

doi:10.1117/12.699718

7 February 2007 Measurement of topographic phase image of living cells by white-light phase-shifting microscope with active stabilization of optical path difference

Toyohiko Yamauchi, Hidenao Iwai, Mitsuharu Miwa, Yutaka Yamashita

Author Affiliations +

Proceedings Volume 6429, Coherence Domain Optical Methods and Optical Coherence Tomography in Biomedicine XI; 64291Q (2007) https://doi.org/10.1117/12.699718
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

We report a full-field, phase-shifting microscope with precise control of the optical path difference (OPD) and show topographic phase images of living cells. Our system is based on a Linnik interference microscope with Kohler illumination of a halogen lamp for the imaging lightsource. Phase-sensitive active-stabilization of the OPD is employed with an infrared laser whose optical path is the same as that of halogen light. We previously reported the results of cultured cell topography with this stabilization scheme; however the phase stability and the image quality were insufficient. We have improved the noise-cancellation system and achieved control of the OPD with stability down to 0.7 nm in the bandwidth of 500 Hz. Quarter wavelength phase-shifting was carried out with sub-nanometer accuracy, and clear topographic phase images of cultured single-layer cells were obtained. The Kohler illumination with the halogen lamp, whose coherence length is 2 &mgr;m, enables homogenous illumination and suppression of artifact signals arising from optical components not associated with the surface of interest.

Citation Download Citation

Toyohiko Yamauchi, Hidenao Iwai, Mitsuharu Miwa, and Yutaka Yamashita "Measurement of topographic phase image of living cells by white-light phase-shifting microscope with active stabilization of optical path difference", Proc. SPIE 6429, Coherence Domain Optical Methods and Optical Coherence Tomography in Biomedicine XI, 64291Q (7 February 2007); https://doi.org/10.1117/12.699718

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