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BioPhotonics Preview - November/December 2022

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Here is your first look at the editorial content for the upcoming November/December issue of BioPhotonics.


Reflectance Confocal Microscopy

Reflectance confocal microscopy is a non-invasive imaging technique that can examine cellular morphologic changes associated with various diseases in vivo. The use of reflectance confocal microscopy could be beneficial, especially in low-resource settings where the equipment, facility, and trained personnel needed for standard histopathologic diagnosis are scarce. However, the widespread use of reflectance confocal microscopy in low-resource settings has not been realized yet due to the high device cost. Recently, low-cost reflectance confocal microscopes that are smartphone-based or smartphone-compatible have been developed. The low-cost confocal devices use inexpensive LEDs, scan-free confocal imaging optics, and consumer-grade 2D imaging sensors, which reduce the overall device cost by an order of magnitude. The low-cost confocal microscopes have been evaluated for cellular imaging of human skin in Uganda and Arizona. Low-cost confocal endomicroscopes have also been developed for imaging internal organs. In the future, the performance of low-cost confocal microscopy will be improved to a level equivalent to that of high-cost confocal microscopes, which will make the low-cost device widely usable in low-resource settings.

Key Technologies: reflectance confocal microscopy, smartphone-based microscopes, LEDs, sensors

Superresolution Microscopy

The breaking of the Abbe limit in fluorescence microscopy in the 1990s carried with it inherent limitations in axial and lateral resolution, which led to the development of various superresolution techniques including stimulated emission depletion (STED), structured illumination microscopy (SIM) and single-molecule localization microscopy (SMLM). In SMLM, which will be the primary focus of this article, individual fluorescent molecules are computationally localized based on their point spread functions (PSFs). One form of SMLM is stochastic optical reconstruction microscopy (STORM), which can visualize molecular organization down to a resolution of 20-30 nm with relatively simple instrumentation, including an inverted microscope frame and an excitation laser(s). This can reveal structural details that provide vital diagnostic clues, such as in cancer detection, where the disruption of heterochromatin is a guidepost of an unstable cellular structure. Commercial SMLM/STORM systems are often marketed to the life science space, especially those studying extracellular vesicles, which are membrane-bound structures that carry protein and other elements in between cells. The more systems are analyzed at this level, the more targeted, effective treatments can be created, and catered to the individual. While these systems are mostly confined to research laboratories, one scientist commented that SMLM systems are only “ one assay away” from being installed in a clinical setting.

Key Technologies: Fluorescence microscopy, superresolution microscopy, single-molecule localization microscopy, stochastic optical reconstruction microscopy, lasers

Scientific Cameras

Scientific cameras and detectors are a vital component of any microscopy- or spectroscopy-based system, affecting the image quality and subsequently the quantitative data acquired in every experiment run on these systems. Scientific cameras have seen many changes and improvements over time, from the earliest CCDs in the 1970’s, to EMCCDs at the turn of the millennium, and further still to the very latest in sCMOS camera technologies in the 2020’s and beyond. What changes or improvements have been made, and how have these advances in camera technology informed changes in microscopes, optics, and even computer systems? This article discusses the past, present, and future of scientific cameras, and how consistent improvements to both hardware and software, and changes to camera sensor architecture have resulted in cameras capable of producing imaging data that would have previously been impossible, capturing unseen events and informing modern science.

Key Technologies: scientific cameras, CCDs, EMCCDs, sCMOS

Dynamic Light Scattering and Alzheimer's

Alzheimers disease is a progressive condition resulting in the loss of mental abilities. During the last 15 years, there have been more than 500 trials of potential therapeutic agents with a failure rate of almost 100%. Most studies lasted 1.5 to 3 years, yet their clinical endpoint – where effectiveness could officially be proven or disproven - required 5 – 10 years.
The early detection of neurologic damage and the tracking of treatment at the microscopic level will reduce research costs and is a prerequisite for the development of potential cures.
Ocular coherence tomography (OCT) images have demonstrated thinning of the retinal nerve fiber and ganglion cell layers in patients with Alzheimer’s disease. Obviously, molecular effects of the condition would lead manifest in changes in imaging of affected systems later.
Dynamic Light Scattering (DLS) measures the scattered light intensity fluctuations resulting from thermal random motion (Brownian motion), and has already been useful in imaging cataracts and diabetes in humans. A clinical retinaI imaging instrument has been developed and in a proof-of-concept study, significant differences in DLS measurements in patients with Alzheimer’s disease were observed, well before other methods captured that information.
The development of an early, noninvasive, quantitative test to diagnose Alzheimer’s disease will lead to differentiating effective drug treatments and hopefully, to the successful therapy before dementia is irreversible.

Key Technologies: OCT, Dynamic Light Scattering, Photo Correlation Spectroscopy

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