Dynamical deformation compensation of phase in digital holographic microscopy

Zikmund, Tomáš; Kvasnica, Lukáš; Uhlířová, Hana; Lovicar, Luděk; Chmelík, Radim

doi:10.1117/12.882171

Cited by 2 publications

(1 citation statement)

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“…The surface is determined from cell‐free regions values. In the first image, the region is specified manually by a human operator (Zikmund et al ., ). In other images of a sequence, it is masked out by fuzzy thresholding.…”

Section: Principle Of the Algorithm For Phase Image Processingmentioning

confidence: 97%

Sequential processing of quantitative phase images for the study of cell behaviour in real‐time digital holographic microscopy

Zikmund

Kvasnica

Týč

et al. 2014

Journal of Microscopy

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Transmitted light holographic microscopy is particularly used for quantitative phase imaging of transparent microscopic objects such as living cells. The study of the cell is based on extraction of the dynamic data on cell behaviour from the time-lapse sequence of the phase images. However, the phase images are affected by the phase aberrations that make the analysis particularly difficult. This is because the phase deformation is prone to change during long-term experiments. Here, we present a novel algorithm for sequential processing of living cells phase images in a time-lapse sequence. The algorithm compensates for the deformation of a phase image using weighted least-squares surface fitting. Moreover, it identifies and segments the individual cells in the phase image. All these procedures are performed automatically and applied immediately after obtaining every single phase image. This property of the algorithm is important for real-time cell quantitative phase imaging and instantaneous control of the course of the experiment by playback of the recorded sequence up to actual time. Such operator's intervention is a forerunner of process automation derived from image analysis. The efficiency of the propounded algorithm is demonstrated on images of rat fibrosarcoma cells using an off-axis holographic microscope.

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Section: Principle Of the Algorithm For Phase Image Processingmentioning

confidence: 97%