Pokroky v digitální holografické mikroskopii: koherencí řízený mikroskop

Advances in Digital Holographic Microscopy: Coherence-Controlled Microscope

conference paper

en

Digital holographic microscope (DHM) allows for direct accessing to the quantitative phase contrast. In this way it becomes a completely non-invasive tool for a contrast living cells observation and the dry-mass density distribution of a cell measurement. Current DHMs use the off-axis holographic setup based on conventional interferometers, which properly work with the coherent illumination. However, this type of illumination is inconvenient for microscopy because of the coherence noise, unwanted interference and a worse lateral resolution. An uncompromising solution to this problem is a DHM based on the Leith achromatic interferometer. While in this setup an off-axis hologram is formed with an arbitrary degree of the illumination coherence, the resolution and the image quality typical for an incoherent-light widefield microscopy can be achieved. Moreover, coherence gating can be introduced which makes the DHM image similar to that of confocal microscope. Hence the character of the DHM image is controlled by the coherence between two extremes: fully coherent-light holography and confocal-microscope-like imaging. The above described possibilities were proved experimentally by the coherence-controlled holographic microscope, which is a DHM based on the achromaticinterferometer. Living cancer cells were observed and their motility was evaluated in the quantitative phase contrast. The presence of the coherence gate was demonstrated by imaging of model samples through a scattering layer.

Digitální holografická mikroskopie (DHM) dává možnost přímo získávat kvantitativní fázový kontrast. Tímto způsobem se stává zcela neinvazivním nástrojem pro kontrastní pozorování živých buněk a měření distribuce hustoty jejich hmoty za sucha. Současné DHM využívají mimoosového uspořádání založeného na konvenčních interferometrech, které správně pracují s koherentním osvětlením. which properly work with the coherent illumination. However, this type of illumination is inconvenient for microscopy because of the coherence noise, unwanted interference and a worse lateral resolution. An uncompromising solution to this problem is a DHM based on the Leith achromatic interferometer. While in this setup an off-axis hologram is formed with an arbitrary degree of the illumination coherence, the resolution and the image quality typical for an incoherent-light widefield microscopy can be achieved. Moreover, coherence gating can be introduced which makes the DHM image similar to that of confocal microscope. Hence the character of the DHM image is controlled by the coherence between two extremes: fully coherent-light holography and confocal-microscope-like imaging. The above described possibilities were proved experimentally by the coherence-controlled holographic microscope, which is a DHM based on the achromaticinterferometer. Living cancer cells were observed and their motility was evaluated in the quantitative phase contrast. The presence of the coherence gate was demonstrated by imaging of model samples through a scattering layer.

Digital holographic microscope (DHM) allows for direct accessing to the quantitative phase contrast. In this way it becomes a completely non-invasive tool for a contrast living cells observation and the dry-mass density distribution of a cell measurement. Current DHMs use the off-axis holographic setup based on conventional interferometers, which properly work with the coherent illumination. However, this type of illumination is inconvenient for microscopy because of the coherence noise, unwanted interference and a worse lateral resolution. An uncompromising solution to this problem is a DHM based on the Leith achromatic interferometer. While in this setup an off-axis hologram is formed with an arbitrary degree of the illumination coherence, the resolution and the image quality typical for an incoherent-light widefield microscopy can be achieved. Moreover, coherence gating can be introduced which makes the DHM image similar to that of confocal microscope. Hence the character of the DHM image is controlled by the coherence between two extremes: fully coherent-light holography and confocal-microscope-like imaging. The above described possibilities were proved experimentally by the coherence-controlled holographic microscope, which is a DHM based on the achromaticinterferometer. Living cancer cells were observed and their motility was evaluated in the quantitative phase contrast. The presence of the coherence gate was demonstrated by imaging of model samples through a scattering layer.

digitální holografie, světelná mikroskopie, holografická mikroskopie, kvantitativní fázový kontrast, koherenční efekty, 3D mikroskopie, zobrazování v kalných prostředích, zobrazování živých buněk

digital holography, light microscopy, holographic microscopy, quantitative phase contrast, coherence effects, three-dimensional microscopy, turbid media imaging, living cells imaging

2011

04.05.2011

SPIE-INT SOC OPTICAL ENGINEERING, 1000 20TH ST, PO BOX 10, BELLINGHAM, WA 98227-0010 USA

BELLINGHAM, USA

9780819486646

0277-786X

Proceedings of SPIE

8074

807402-1–807402-7

328