Microscopy with ultraviolet surface excitation for rapid slide-free histology.

Histologic examination of tissues is central to the diagnosis and management of neoplasms and many other diseases, and is a foundational technique for preclinical and basic research. However, commonly used bright-field microscopy requires prior preparation of micrometre-thick tissue sections mounted on glass slides, a process that can require hours or days, that contributes to cost, and that delays access to critical information. Here, we introduce a simple, non-destructive slide-free technique that within minutes provides high-resolution diagnostic histological images resembling those obtained from conventional haematoxylin-and-eosin-histology. The approach, which we named microscopy with ultraviolet surface excitation (MUSE), can also generate shape and colour-contrast information. MUSE relies on ~280-nm ultraviolet light to restrict the excitation of conventional fluorescent stains to tissue surfaces, and it has no significant effects on downstream molecular assays (including fluorescence in situ hybridization and RNA-seq). MUSE promises to improve the speed and efficiency of patient care in both state-of-the-art and low-resource settings, and to provide opportunities for rapid histology in research

Mid-infrared OCT imaging in highly scattering samples using real-time upconversion of broadband supercontinuum covering from 3.6-4.6 μm

We present a mid-infrared spectral-domain optical coherence tomography system operating at 4.1 μm central wavelength with a high axial resolution of 8.6 μm enabled by more than 1 μm bandwidth from 3.58-4.63 μm produced by a mid-infrared supercontinuum laser. The system produces 2D cross-sectional images in real-time enabled the high-brightness of the supercontinuum source in combination with broadband upconversion of the signal to the range 820-865 nm, where a standard 800 nm array spectrometer can be used for fast detection. We discuss the potential applications within nondestructive testing in highly scattering materials and within biomedical imaging for achieving the in-vivo optical biopsy

Origins of Optical Absorption Characteristics of Cu²⁺ Complexes in Aqueous Solutions

Many transition metal complexes exhibit infrared or visible optical absorption arising from d-d transitions that are the key to functionality in technological applications and biological processes. The observed spectral characteristics of the absorption spectra depend on several underlying physical parameters whose relative contributions are still not fully understood. Although conventional arguments based on ligand-field theory can be invoked to rationalize the peak absorption energy, they cannot describe the detailed features of the observed spectral profile such as the spectral width and shape, or unexpected correlations between the oscillator strength and absorption peak position. Here, we combine experimental observations with first-principles simulations to investigate origins of the absorption spectral profile in model systems of aqueous Cu2+ ions with Cl-, Br-, NO2- and CH3CO2- ligands. The ligand identity and concentration, fine structure in the electronic d-orbitals of Cu2+, complex geometry, and solvation environment are all found to play key roles in determining the spectral profile. Moreover, similar physiochemical origins of these factors lead to interesting and unexpected correlations in spectral features. The results provide important insights into the underlying mechanisms of the observed spectral features and offer a framework for advancing the ability of theoretical models to predict and interpret the behavior of such systems

Towards real-Time non contact spatial resolved oxygenation monitoring using a multi spectral filter array camera in various light conditions

Non contact spatial resolved oxygenation measurements remain an open challenge in the biomedical field and non contact patient monitoring. Although point measurements are the clinical standard till this day, regional differences in the oxygenation will improve the quality and safety of care. Recent developments in spectral imaging resulted in spectral filter array cameras (SFA). These provide the means to acquire spatial spectral videos in real-Time and allow a spatial approach to spectroscopy. In this study, the performance of a 25 channel near infrared SFA camera was studied to obtain spatial oxygenation maps of hands during an occlusion of the left upper arm in 7 healthy volunteers. For comparison a clinical oxygenation monitoring system, INVOS, was used as a reference. In case of the NIRS SFA camera, oxygenation curves were derived from 2-3 wavelength bands with a custom made fast analysis software using a basic algorithm. Dynamic oxygenation changes were determined with the NIR SFA camera and INVOS system at different regional locations of the occluded versus non-occluded hands and showed to be in good agreement. To increase the signal to noise ratio, algorithm and image acquisition were optimised. The measurement were robust to different illumination conditions with NIR light sources. This study shows that imaging of relative oxygenation changes over larger body areas is potentially possible in real time

Principal component model of multispectral data for near real-time skin chromophore mapping

Multispectral images of skin contain information on the spatial distribution of biological chromophores, such as blood and melanin. From this, parameters such as blood volume and blood oxygenation can be retrieved using reconstruction algorithms. Most such approaches use some form of pixelwise or volumetric reconstruction code. We explore the use of principal component analysis (PCA) of multispectral images to access blood volume and blood oxygenation in near real time. We present data from healthy volunteers under arterial occlusion of the forearm, experiencing ischemia and reactive hyperemia. Using a two-layered analytical skin model, we show reconstruction results of blood volume and oxygenation and compare it to the results obtained from our new spectral analysis based on PCA. We demonstrate that PCA applied to multispectral images gives near equivalent results for skin chromophore mapping and quantification with the advantage of being three orders of magnitude faster than the reconstruction algorithm