BioPhotonics Preview - November/December 2023

Live Cell Imaging

Live cell imaging, powered by AI and advanced microscopes, allows the study of dynamic processes, behavior and functions over time. It is particularly useful when an endpoint is unknown, for example in the long-term study of disease progression or pharmaceutical treatments. The largest segment of this growing market is time-lapse microscopy, which has fueled a number of recent developments. For example, the Bright Eyes time-tagging module, developed in the EU, is rooted in FPGAs that can tag single-photon events, capturing them in real time in multiple dimensions (such as protein aggregation). And line-scanning Brillouin microscopy uses light’s interactions with thermal vibrations in a cell to capture mechanics over space and time. A number of companies are working on advanced software which provides automated z-stacking to reconstruct images from 2D into 3D noninvasively. Other companies are marketing specialized illumination that can help improve imaging speed with a minimum of phototoxicity. Drug discovery, cancer research and neuroscience are some of the key areas that are fueling this innovation and institutions and companies in that space will continue to dominate the live cell imaging market.

Key Technologies: AI, time-lapse microscopy

Light-Sheet Microscopy

Light sheet microscopy, also known as selective plane illumination microscopy (SPIM), is a powerful imaging technique used in a wide range of research fields. It is characterised by a thin sheet of laser light illuminating a specimen; simultaneously, the fluorescent information is captured by an uncoupled detection objective perpendicular to the imaging axis. Understanding whether SPIM is the ideal choice can be a dilemma for many scientists. In this practical article, a diverse team, spanning from biologists to physicists and end-users to microscope developers, collaborates to provide a comprehensive practical guide. We will examine the advantages of SPIM imaging, explore geometries, address the data processing challenges, and provide a brief insight into the future of SPIM imaging. Together, whether you seek practical hands-on insights or aim to grasp the potential of SPIM, this article is aimed as practical resource.

Key Technologies: Light Sheet Microscopy, Selective Plane Illumination Microscopy, lasers

Fluorescence Imaging

Necrotizing soft-tissue infections (NSTIs), commonly known as “flesh-eating bacteria,” are fast-moving and deadly. They occur when virulent bacteria infect fascia, the connective tissue that surrounds all structures in the body. Fascia provides a permissive environment for rapid bacterial advancement, and within hours, can lead to sepsis, multi-organ failure, and death. Standard-of-care has not improved substantially for decades: prompt, aggressive surgical removal of affected tissues and broad-spectrum antibiotics. Patients with NSTIs commonly present with non-specific symptoms, leading to frequent misdiagnoses and delays in care. NSTIs cause prominent vascular clotting in affected tissues. In this study, indocyanine green (ICG), a safe, vascular perfusion fluorophore, is intravenously administered to patients suspected of having NSTIs. Within seconds, ICG enables visual demarcation of local vasculature. Patients retrospectively confirmed to have NSTIs have distinct fluorescent signal voids in affected tissues, indicative of clotting caused by the infection. ICG imaging may expedite rapid identification and treatment of NSTIs, leading to improvements in patient outcomes.

Key Technologies: Fluorescence Imaging, ICG

Lasers and Multiphoton Microscopy

The ultrafast laser is the critical component underpinning all multiphoton excitation (MPE) microscopy methods. So it’s not surprising that the evolution of MPE microscopy is accompanied and enabled by a corresponding evolution in ultrafast laser sources. In this article, We will highlight three different areas of MPE microscopy and describe the key characteristics of the associated lasers. These are (a) In-vivo tissue imaging, particularly in neuroscience, where imaging depth, speed and versatility are important; (b) Pre-clinical and specialized applications where compact fixed wavelength lasers can be conveniently fully integrated into instrumentation; (c) emerging applications like three-photon excitation, which needs higher power at longer wavelengths.

Key Technologies: Ultrafast lasers, multiphoton microscopy

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