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Published February 1, 2021 | Supplemental Material + Published
Journal Article Open

Using the Gouy phase anomaly to localize and track bacteria in digital holographic microscopy 4D images

Abstract

Described over 100 years ago, the Gouy phase anomaly refers to the additional π phase shift that is accumulated as a wave passes through focus. It is potentially useful in analyzing any type of phase-sensitive imaging; in light microscopy, digital holographic microscopy (DHM) provides phase information in the encoded hologram. One limitation of DHM is the weak contrast generated by many biological cells, especially unpigmented bacteria. We demonstrate here that the Gouy phase anomaly may be detected directly in the phase image using the z-derivative of the phase, allowing for precise localization of unlabeled, micrometer-sized bacteria. The use of dyes that increase phase contrast does not improve detectability. This approach is less computationally intensive than other procedures such as deconvolution and is relatively insensitive to reconstruction parameters. The software is implemented in an open-source FIJI plug-in.

Additional Information

© 2020 Optical Society of America. Provided under the terms of the OSA Open Access Publishing Agreement. Received 4 August 2020; revised 9 October 2020; accepted 19 October 2020; posted 22 October 2020 (Doc. ID 404004); published 10 November 2020. Part of this work was carried out at, or under a contract with, the Jet Propulsion Laboratory, California Institute of Technology, under contract with NASA. We thank J. Kent Wallace and Louis Sumrall for useful discussions. The FeatureJ plug-ins were written by Erik Meijering. Funding: Jet Propulsion Laboratory; National Science Foundation (1828793). The authors declare no conflicts of interest.

Attached Files

Published - josaa-38-2-A11.pdf

Supplemental Material - 4880204.pdf

Supplemental Material - Visualization1.mp4

Supplemental Material - Visualization2.mp4

Supplemental Material - Visualization3.mp4

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August 20, 2023
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