Abstract
.A coherence-controlled holographic microscope (CCHM) was developed particularly for quantitative phase imaging and measurement of live cell dynamics, which is the proper subject of digital holographic microscopy (DHM). CCHM in low-coherence mode extends DHM in the study of living cells. However, this advantage is compensated by sensitivity of the system to easily become misaligned, which is a serious hindrance to wanted performance. Therefore, it became clear that introduction of a self-correcting system is inevitable. Accordingly, we had to devise a theory of a suitable control and design an automated alignment system for CCHM. The modulus of the reconstructed holographic signal was identified as a significant variable for guiding the alignment procedures. From this, we derived the original basic realignment three-dimensional algorithm, which encompasses a unique set of procedures for automated alignment that contains processes for initial and advanced alignment as well as long-term maintenance of microscope tuning. All of these procedures were applied to a functioning microscope and the tested processes were successfully validated. Finally, in such a way, CCHM is enabled to substantially contribute to study of biology, particularly of cancer cells in vitro.
Highlights
The coherence-controlled holographic microscope (CCHM)[1,2] is an innovative system designed for quantitative phase imaging (QPI) and measurement of live cell dynamics.[3]
CCHM works with a broad polychromatic light source, which makes all the difference to other off-axis holographic microscopes usually equipped with a laser light source.[8,9]
We have developed a unique set of procedures constituting the original basic realignment three-dimensional algorithm (BReTA)
Summary
The coherence-controlled holographic microscope (CCHM)[1,2] is an innovative system designed for quantitative phase imaging (QPI) and measurement of live cell dynamics.[3]. Low coherence improves lateral resolution and the imaging in general.[7] For exploiting these effects, CCHM works with a broad polychromatic light source, which makes all the difference to other off-axis holographic microscopes usually equipped with a laser light source.[8,9] This is because the high-coherence light source leads to the formation of unwanted artifacts in QPI as a consequence of coherence noise, random interferences, and diffraction of light. Low-coherence illumination, requires precise alignment and high stability of the system to be maintained during long-lasting timelapse QPI studies of activity and reaction of living cells. Just these advantages stand for the contribution of CCHM to cell biology research. The interferometer state detection is carried out by an auxiliary detector[10]
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