We present spatial light interference microscopy (SLIM) as a new optical microscopy technique, capable of measuring nanoscale structures and dynamics in live cells via interferometry. SLIM combines two classic ideas in light imaging: Zernike’s phase contrast microscopy, which renders high contrast intensity images of transparent specimens, and Gabor’s holography, where the phase information from the object is recorded. Thus, SLIM reveals the intrinsic contrast of cell structures and, in addition, renders quantitative optical path-length maps across the sample. The resulting topographic accuracy is comparable to that of atomic force microscopy, while the acquisition speed is 1,000 times higher. We illustrate the novel insight into cell dynamics via SLIM by experiments on primary cell cultures from the rat brain. SLIM is implemented as an add-on module to an existing phase contrast microscope, which may prove instrumental in impacting the light microscopy field at a large scale.
The refractive index of human skin tissues is an important parameter in characterizing the optical response of the skin. We extended a previously developed method of coherent reflectance curve measurement to determine the in vitro values of the complex refractive indices of epidermal and dermal tissues from fresh human skin samples at eight wavelengths between 325 and 1557 nm. Based on these results, dispersion relations of the real refractive index have been obtained and compared in the same spectral region.
Fourier transform light scattering (FTLS) is a novel experimental approach that combines optical microscopy, holography, and light scattering for studying inhomogeneous and dynamic media. In FTLS the optical phase and amplitude of a coherent image field are quantified and propagated numerically to the scattering plane. Because it detects all the scattered angles (spatial frequencies) simultaneously in each point of the image, FTLS can be regarded as the spatial equivalent of Fourier transform infrared spectroscopy, where all the temporal frequencies are detected at each moment in time.
We constructed an automated reflectometry system for accurate measurement of coherent reflectance curves of turbid samples and analyzed the presence of coherent and diffuse reflection near the specular reflection angle. An existing method has been validated to determine the complex refractive indices of turbid samples on the basis of nonlinear regression of the coherent reflectance curves by Fresnel's equations. The complex refractive indices of fresh porcine skin epidermis and dermis tissues and Intralipid solutions were determined at eight wavelengths: 325, 442, 532, 633, 850, 1064, 1310, and 1557 nm.
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