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Journal of the Optical Society of America A

Journal of the Optical Society of America A


  • Vol. 9, Iss. 6 — Jun. 1, 1992
  • pp: 924–931

Separation of phase and absorption images in phase-contrast microscopy

Tomoya Noda and Satoshi Kawata  »View Author Affiliations

JOSA A, Vol. 9, Issue 6, pp. 924-931 (1992)

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A method for separating the phase distribution from the image of a semitransparent sample observed with a phase-contrast microscope is proposed. Three images of the same sample are used for calculating the phase distribution, one observed by a normal objective lens and the others observed by a bright-contrast objective and a dark-contrast objective. The method works even for samples that are not weak phase objects. The phase (or refractive-index) distribution is quantitatively reconstructed by numerically solving a set of linear equations for every pixel of the sample image. The method of phase reconstruction is mathematically derived and is explained with graphs in the complex amplitude space. Experimental results are also presented.

© 1992 Optical Society of America

Original Manuscript: July 23, 1991
Revised Manuscript: December 5, 1991
Manuscript Accepted: December 6, 1991
Published: June 1, 1992

Tomoya Noda and Satoshi Kawata, "Separation of phase and absorption images in phase-contrast microscopy," J. Opt. Soc. Am. A 9, 924-931 (1992)

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  1. F. Zernike, “Das Phasenkontrastverfahren bei der mikroskopischen Beobachtung,”Z. Tech. Phys. 16, 454–457 (1935).
  2. J. W. Goodman, Introduction to Fourier Optics (McGraw-Hill, New York, 1968).
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  4. W. G. Driscoll, W. Vaughan, Handbook of Optics (McGraw-Hill, New York, 1978), Chap. 6.
  5. W. Lang, Nomarski Differential Interference-Contrast Microscopy (reprint: Zeiss, 7082 Oberkochen, Germany, 1975).

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