System for coherence-controlled holographic microscopy of living cells

System for coherence-controlled holographic microscopy of living cells

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Coherence Controlled Holographic Microscopy (CCHM) is a novel holographic technique for quantitative-phase contrast (QPC) biological observations particularly of living cells. Owing to the ordinary (low coherence) illumination source, the CCHM images are of low noise, deprived of coherence noise (speckles) and the lateral resolution is improved by a factor of 2 compared to classic holographic microscopes. Long-lasting time-lapse experiments require elimination of the CCHM optical system instability in order to achieve precise QPC measurement and to maintain correct CCHM adjustment for its low-coherence operation. The critical part of CCHM is the interferometer, which is very sensitive to temperature fluctuations and air turbulences. The temperature stabilization of the whole microscope without air turbulences is therefore required to provide stability for long-term observations of living cells. Novel heated microscope box and stage designed and constructed for this purpose are described in the paper. The system maintains a constant temperature of both the microscope and of the sample set to 37 deg thus providing optimal living conditions for living human and animal cells. The system is completed with a novel flow-chamber for living-cells accommodation during observation. A service of the system to CCHM is demonstrated by a series of pictures of growing cells.

Coherence Controlled Holographic Microscopy (CCHM) is a novel holographic technique for quantitative-phase contrast (QPC) biological observations particularly of living cells. Owing to the ordinary (low coherence) illumination source, the CCHM images are of low noise, deprived of coherence noise (speckles) and the lateral resolution is improved by a factor of 2 compared to classic holographic microscopes. Long-lasting time-lapse experiments require elimination of the CCHM optical system instability in order to achieve precise QPC measurement and to maintain correct CCHM adjustment for its low-coherence operation. The critical part of CCHM is the interferometer, which is very sensitive to temperature fluctuations and air turbulences. The temperature stabilization of the whole microscope without air turbulences is therefore required to provide stability for long-term observations of living cells. Novel heated microscope box and stage designed and constructed for this purpose are described in the paper. The system maintains a constant temperature of both the microscope and of the sample set to 37 deg thus providing optimal living conditions for living human and animal cells. The system is completed with a novel flow-chamber for living-cells accommodation during observation. A service of the system to CCHM is demonstrated by a series of pictures of growing cells.

Holographic microscopy, temperature control, culture chambers

Holographic microscopy, temperature control, culture chambers

ANTOŠ, M.; ČOLLÁKOVÁ, J.; VESELÝ, P.; CHMELÍK, R.; KŘÍŽOVÁ, A.

2013

18.12.2012

SPIE

978-80-244-3408-7

Proceedings of SPIE

0277-786X

Proceedings of SPIE

8697

Spojené státy americké

869712-1

869712-6

6