Phase measurement using DIC microscopy

Koos Krisztián and Peksel Begüm and Kelemen Lóránd: Phase measurement using DIC microscopy. In: Acta cybernetica, (23) 2. pp. 629-643. (2017)

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The development of fluorescent probes and proteins has helped make light microscopy more popular by allowing the visualization of specific subcellular components, location and dynamics of biomolecules. However, it is not always feasible to label the cells as it may be phototoxic or perturb their functionalities. Label-free microscopy techniques allow us to work with live cells without perturbation and to evaluate morphological differences, which in turn can provide useful information for high-throughput assays. In this study, we use one of the most popular label-free techniques called differential interference contrast (DIC) microscopy to estimate the phase of cells and other nearly transparent objects and instantly estimate their height. DIC images provide detailed information about the optical path length (OPL) differences in the sample and they are visually similar to a gradient image. Our previous DIC construction algorithm outputs an image where the values are proportional to the OPL (or implicitly the phase) of the sample. Although the reconstructed images are capable of describing cellular morphology and to a certain extent turn DIC into a quantitative technique, the actual OPL has to be computed from the input DIC image and the microscope calibration settings. Here we propose a computational method to measure the phase and approximate height of cells after microscope calibration, assuming a linear formation model. After a calibration step the phase of further samples can be determined when the refractive indices of the sample and the surrounding medium is known. The precision of the method is demonstrated on reconstructing the thickness of known objects and real cellular samples.

Item Type: Article
Journal or Publication Title: Acta cybernetica
Date: 2017
Volume: 23
Number: 2
ISSN: 0324-721X
Page Range: pp. 629-643
Language: English
Place of Publication: Szeged
Related URLs:
DOI: 10.14232/actacyb.23.2.2017.12
Uncontrolled Keywords: Képfeldolgozás - digitális, Képszerkesztés, Mikroszkópia
Additional Information: Bibliogr.: p. 641-643. ; ill. ; összefoglalás angol nyelven
Subjects: 01. Natural sciences
01. Natural sciences > 01.02. Computer and information sciences
Date Deposited: 2018. Feb. 13. 10:23
Last Modified: 2022. Jun. 20. 15:16

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