Bile duct cancer, or cholangiocarcinoma, is a prevalent liver cancer often diagnosed at advanced stages, leading to poor survival rates. Therefore, the development of a reliable early detection technique is urgently needed. Current imaging techniques lack the necessary accuracy to distinguish between dysplastic and benign biliary ducts. Endoscopic techniques, while capable of directly assessing the bile duct lining, often suffer from insufficient sampling. In this paper we discuss a novel endoscopic optical light scattering technique designed to evaluate the malignant potential of the bile duct. The technique employs an ultraminiature spatial gating fiber optic probe compatible with cholangioscopes and endoscopic retrograde cholangiopancreatography (ERCP) catheters. The miniature optical probe enables the detailed investigation of the internal cellular composition of the bile duct epithelium using light scattering spectroscopy (LSS) and also allows for the assessment of the phenotypic properties of the underlying connective tissue with diffuse reflectance spectroscopy (DRS). In a pilot in vivo double-blind prospective study involving 29 patients undergoing routine ERCP procedures, the technique detected malignant transformation with 97% accuracy. Our pilot study suggests that biliary duct pre-cancer can be identified non-invasively in vivo, offering a promising new avenue for early detection and intervention in bile duct cancer.
Coherent confocal light absorption and scattering spectroscopic (C-CLASS) microscopy, which extends the principles of light scattering spectroscopy to subcellular imaging, can be used to reveal biological structures well beyond the diffraction limit. Here we show that high-resolution C-CLASS microscopy can be used to detect nanoscale changes in chromatin structure. Unlike most methods for chromatin monitoring, C-CLASS microscopy can be used label-free in live cells. Live differentiating hiPSC organoids were measured over the space of sixteen days and characteristic chromatin changes were observed.
The COVID-19 pandemic has caused a marked disruption in the delivery of medical care, resulting in significant negative consequences for patients. Considering Covid-19 spreads primarily through expelled respiratory droplets, the ability to detect and measure droplets is critical to the development of clinical protective practices. However, most available methods are either unsuitable for the clinical setting, or cannot distinguish solid particles from liquid droplets. We developed a robust and portable optical instrument capable of measuring the size and quantity of droplets generated during medical procedures. Here we outline the system design and describe our preclinical measurements, which showed that surgical masks significantly reduce the number of expelled speech droplets.
Nanoscale changes in the nuclear structure have been shown to play a critical role in genetic and transcriptional alterations and are a hallmark of neoplasia. Genomic processes are regulated by chromatin packing density, thus underlying the significance of understanding the subnuclear structure and its role in the regulation of molecular processes. However, the dynamic and multiscale aspects of these phenomena have remained an open problem. The key reason is the lack of technologies for label-free nanoscale-sensitive measurements in live cells. We have developed confocal light absorption and scattering spectroscopic (CLASS) microscopy for label-free chromatin sensing in live cells.
In this talk, we will discuss how scattered light can be used for noninvasive detection of invisible pre-cancer in organs such as the esophagus or pancreas which seem to have little in common. Nevertheless, since pre-cancer in many organs is characterized by certain common microscopic changes in the epithelial cells, such as the increase in nuclear size and nuclear density, we will show that light scattering signatures of those pre-cancers are quite similar, allowing for early cancer imaging and detection without the need for external markers. Light scattering signatures can also be used for sensing subnuclear and subcelluar structures, such as chromatin packing, organelle organization, and characterization of cell-derived exosomes. Nanoscale changes in the nuclear structure have been shown to play a critical role in genetic and transcriptional alterations and are a hallmark of neoplasia. We will discuss how the approach based on the combination of confocal microscopy and spectroscopy
We have constructed a Holographic Scanning Microscopy (HSM) setup, which employs an optical design and a scanning unit of a commercial confocal laser scanning microscope. This arrangement helps to construct the HSM easier, but the scanning unit gives additional phase distortions modifying an object wave and introducing phase shift. We offer a way to dispose such distortions changing the way of recording holograms and reconstruction algorithm. Analyzing recorded holograms we conclude, that we need to record a hologram of flat mirror first, which would contain required information about phase distortions, and use it for reconstruction. Finally reconstruction algorithm becomes like to “coded” reference wave reconstruction algorithm. Proposed algorithm was successfully implemented and tested in MatLab.
Holographic scanning microscopy - novel technique both in laser scanning microscopy and digital holographic microscopy allow multimodal approach to cell and tissue investigation in biomedical applications promising new advantages (quantitative phase imaging, superresolution, computerized tomography), but regular reconstruction leads to incorrectness. Analysis of light propagation through the schematics allows to offer reconstruction procedures depending on recording conditions.
Optical methods for study biological tissue and cell at micro- and nanoscale level step now over diffraction limit. Really it is single molecule localization techniques that achieve the highest spatial resolution. One of those techniques, called bleaching/blinking assisted localization microscopy (BaLM) relies on the intrinsic bleaching and blinking behavior characteristic of commonly used fluorescent probes. This feature is the base of BaLM image series acquisition and data analysis. In our work blinking of single fluorescent spot against a background of others comes to light by subtraction of time series successive frames. Then digital estimation gives the center of the spot as a point of fluorescent molecule presence, which transfers to other image with higher resolution according to accuracy of the center localization. It is a part of image with improved resolution. This approach allows overlapping fluorophores and not requires single photon sensitivity, so we use 8,8 megapixel CMOS camera with smallest (1.55 um) pixel size. This instrumentation on the base of Zeiss Axioscope 2 FS MOT allows image transmission from object plane to matrix on a scale less than 100 nm/pixel using 20x-objective, thereafter the same resolution and 5 times more field of view as compared to EMCCD camera with 6 um pixel size. To optimize excitation light power, frame rate and gain of camera we have made appropriate estimations taking into account fluorophores behaviors features and equipment characteristics. Finely we have clearly distinguishable details of the sample in the processed field of view.
One of the main constituent parts of optical coherent measuring apparatus is laser as source with stable performance of
frequency, radiation intensity, and light beam uniformity. At present time semiconductor lasers are rather attractive devices
in view of there low prices, small size, serviceability. Progress in its quality leads to including them not only in lightheads,
but as lighting unit in measuring apparatus. In order to guarantee accuracy of measuring instruments, all parts of them must
have stable performance, and in this respect semiconductor laser demand stabilization more that one characteristic quantity
at once. And frequency stability on the one hand is overwhelmingly important for constancy of optical measuring
instruments, on the other hand our investigations show that its regulatory control is very arduous task. Both holographic
methods and phase modulated speckle interferometry clearly recognize smooth frequency shift and frequency jumping
depending on pumping current and temperature. And for repeatability it's required to return both of them. So it is necessary
laser frequency testing during working. For interferometric comparison circuit it is frequency variation that exerts influence
on fringes pattern generation, so just this parameter should be traced in the course of measuring. Specially prepared test
object, introduced in holographic scheme, allows to uncover frequency variation, if they had have place, and to reproduce
coherence function of laser source. Complicated coherence function of semiconductor lasers can destroy interference pattern
or foul the interpretation of it. So this coherence calibration is also very useful for results validity. Phase modulated speckle
interferometry method allows to build phase correlation portraits, analogical to interferograms, hence multiwavelength
contour generation masks the picture of intrinsic object information too. Both real wavelength change and nonresolution
area, when coherence length is less then path-length difference, may be obtained with the help of known wedge incline near
the measured object. Since measured and testing image will be entered at the same time, test results will be applied in
interpretation of measurement results. Thus on-line testing of laser characteristic quantities allows using unstable sources
and increasing of measurement precision.
Mode structure of semiconductor lasers may be scrutinized closely by reconstructed hologram image analysis if one use
specially prepared test object. On the other hand specially selected mode structure of illumination enables to test surface
condition. We make good use of Lippmann-Bragg holography setup to achieve these purposes.
Three-dimensional image creation based on registration interference pattern of Fizeau interferometer is considered. Recording of interference pattern allows storing both intensity and direction of propagation of incoherent light waves. It gives the possibility of wave front reconstruction without reference wave. 3D displays compatible to usual may be made. Optical experiment proves this possibility and estimations for the computer display construction is done.
The registration of an autocorrelation function of an object field with the help of Fizeau interferometer was carried out in order to reconstruct an object field and derive the volumetric image without information redundancy. In result the obtained image corresponded expected.
Fizeau interferometer pattern analysis have carried out in the case of arbitrary slope beam illumination. Analysis get localization plane location and its dependence on wedge relative index of refraction and on the interferometer entrance mirror slope angle to the incident wave. The analyses fits experiment.
Fizeau interferometer pattern analysis have carried out in the case of arbitrary slope beam illumination. Analysis get localization plane location (that is in coincidence with image plane) and its dependence on wedge relative index of refraction and on the interferometer entrance mirror slope angle to the incidence wave. The analyses fits experiment.
Equipment for image holography and for holographic nondestructive testing on the base of ruby laser was designed and constructed. Large coherence length in free- lasing regime is reached.
Fabry-Perot interferometer outline analysis have carried out in the case of wide fan beam illumination. The analyses fits experiment. Holographic setup optimization with this results is recommended.
Diffuse surface relief profile is obtained by contour map generation in Lippmann-Bragg scheme at basic and doubled frequencies of ruby laser. Results demonstrate possibilities of realized experimental setup for measuring and testing of diffuse surfaces relief.
Application of Ti:sapphire femtosecond laser for 3-D image holographic recording of superfast processes in its temporal development is offered. Pulse shortening leads to time resolution improvement of the systems.
Diffuse surface relief profile is obtained by contour map generation with the help of multiple- wavelength holography at basic and doubled frequencies of ruby laser. Correlation with etalon may also be produced.
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