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
Gold nanorods have the potential to be employed as extremely bright molecular marker labels. However, samples
containing a large number of gold nanorods usually exhibit relatively wide spectral lines. This linewidth limits the use of
the nanorods since it would be rather difficult to image several types of nanorod markers simultaneously. We measured
native scattering spectra of single gold nanorods with the CLASS microscope and found that single gold nanorods have a
narrow spectrum as predicted by the theory. That suggests that nanorod-based molecular markers with controlled narrow
aspect ratios should provide spectral lines sufficiently narrow for effective biomedical imaging.
We have developed a novel optical method for observing submicron intracellular structures in living cells
which is called confocal light absorption and scattering spectroscopic (CLASS) microscopy. It combines
confocal microscopy, a well-established high-resolution microscopic technique, with light scattering
spectroscopy (LSS). CLASS microscopy requires no exogenous labels and is capable of imaging and
continuously monitoring individual viable cells, enabling the observation of cell and organelle functioning at
scales on the order of 100 nm. In addition, it provides not only size information but also information about the
biochemical and physical properties of the cell.
Confocal light absorption and scattering spectroscopic (CLASS) microscopy is a novel optical technique for observing
submicron intracellular structures in living cells. It allows monitoring nondestructively cell function and cell dynamics in
vivo and in real time. CLASS microscopy, having accuracy well beyond the diffraction limit, does not require cell
fixation as the electron microscopy. In addition, it provides not only size information but also information about the
biochemical and physical properties of the cell. CLASS microscopy can also visualize multiple compartments inside of
living cell without employing exogenous molecular markers which are required by fluorescence microscopy and which
can affect normal cell functioning. Recently we improved our CLASS microscope by utilizing the full power output of
the supercontinuum laser and used it to study apoptosis in live cells.
Le Qiu, Hui Fang, Edward Vitkin, Munir Zaman, Charlotte Andersson, Saira Salahuddin, Lauren Kimerer, Patsy Cipolloni, Mark Modell, Steven Freedman, Irving Bigio, Irving Itzkan, Eugene Hanlon, Lev Perelman
We recently developed a novel optical method for observing submicron intracellular structures in living cells which is
called confocal light absorption and scattering spectroscopic (CLASS) microscopy. It combines confocal microscopy, a
well-established high-resolution microscopic technique, with light scattering spectroscopy (LSS). CLASS microscopy
requires no exogenous labels and is capable of imaging and continuously monitoring individual viable cells, enabling the
observation of cell and organelle functioning at scales on the order of 100 nm with 10 nm accuracy. To demonstrate the
ability of the CLASS microscope to monitor unstained living cells on submicrometer scale we studied human bronchial
epithelial cells undergoing apoptosis. Fluorescence microscopy of living cells requires application of molecular markers
which can affect normal cell functioning. CLASS microscopy is not affected by this avoiding potential interference of
fluorescence molecular markers with cell processes. In addition, it provides not only size information but also
information about the biochemical and physical properties of the cell. CLASS microscopy can provide unique
capabilities for the study of cell interactions with the environment, cell reproduction and growth and other functions of
viable cells, which are inaccessible by other techniques.
Le Qiu, Edward Vitkin, Hui Fang, Munir Zaman, Charlotte Andersson, Saira Salahuddin, Mark Modell, Steven Freedman, Eugene Hanlon, Irving Itzkan, Lev Perelman
We recently developed a new microscopic optical technique capable of noninvasive analysis of cell structure and cell
dynamics on the submicron scale [1]. It combines confocal microscopy, a well-established high-resolution microscopic
technique, with light scattering spectroscopy (LSS) and is called confocal light absorption and scattering spectroscopic
(CLASS) microscopy. CLASS microscopy requires no exogenous labels and is capable of imaging and continuously
monitoring individual viable cells, enabling the observation of cell and organelle functioning at scales on the order of
100 nm.
To test the ability of CLASS microscopy to monitor cellular dynamics in vivo we performed experiments with human
bronchial epithelial cells treated with DHA and undergoing apoptosis. The treated and untreated cells show not only
clear differences in organelle spatial distribution but time sequencing experiments on a single cell show disappearance of
certain types of organelles and change of the nuclear shape and density with the progression of apoptosis.
In summary, CLASS microscopy provides an insight into metabolic processes within the cell and opens doors for the
noninvasive real-time assessment of cellular dynamics. Noninvasive monitoring of cellular dynamics with CLASS
microscopy can be used for a real-time dosimetry in a wide variety of medical and environmental applications that have
no immediate observable outcome, such as photodynamic therapy, drug screening, and monitoring of toxins.
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