Publication | Open Access
High-resolution quantitative phase-contrast microscopy by digital holography
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2005
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HolographyEngineeringCoherent NoiseMedical ImagingMicroscopyAngular Spectrum MethodNovel Imaging MethodBiomedical ImagingQuantitative Phase ImagingBiomedical EngineeringHolographic MethodLight MicroscopyMedicineCell BiologyBiophysicsDigital Holography
The study aims to improve digital holography for high‑resolution, high‑fidelity quantitative phase‑contrast microscopy. The authors enhance digital holography techniques, notably using the angular spectrum method, to achieve these images. The angular spectrum method produces holographic phase images with 0.5 µm diffraction‑limited resolution, 30 nm optical‑thickness accuracy, low coherent noise, and clear visualization of organelles in SKOV‑3 ovarian cancer cells.
Techniques of digital holography are improved in order to obtain high-resolution, high-fidelity images of quantitative phase-contrast microscopy. In particular, the angular spectrum method of calculating holographic optical field is seen to have significant advantages including tight control of spurious noise components. Holographic phase images are obtained with 0.5 mum diffraction-limited lateral resolution and largely immune from the coherent noise common in other holographic techniques. The phase profile is accurate to about 30 nm of optical thickness. Images of SKOV-3 ovarian cancer cells display intracellular and intranuclear organelles with clarity and quantitative accuracy.