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Advancement of phase-contrast microscopy (Neuromethods 2018)


Diffraction phenomena governing image formation in the microscope were employed to improve the performance of phase-contrast microscopy.

Diffractograms of highly periodic objects (B) are prominent at the objective back focal plane (insets in A and B).

A. With no diffracting objects in the viewing field only the condenser (illuminating) diaphragm can be seen.

B. Periodic ribs in the body scales of silverfish (Lepisma saccharina). Depending on their density (spatial frequency) and orientation the central bright annulus (image of the condenser diaphragm) is split into several eccentric annuli.

C. Living unstained buccal epithelial cells isolated from one of the authors (RP). Apodized phase-contrast microscopy (invented by Tatsuro Otaki 1999) reduces ‘halo‘ artifacts but also impairs image contrast in optically thin cells.

Other improvements of this kind are described, suiting different cell types.

Adapted from:

Pelc R., Hostounský Z., Otaki T. & Katoh K. (2018 in press) Conventional, apodized and relief phase-contrast microscopy. In: Neurohistology & Imaging: Basic Techniques (Pelc R., Doucette J.R. & Walz W., Eds.), pp. XXX-XXX. Springer (Humana Press), New York (the NEUROMETHODS series, vol. XX)