In vivo simultaneous nonlinear absorption Raman and fluorescence (SNARF) imaging of mouse brain cortical structures.

Label-free multiphoton microscopy is a powerful platform for biomedical imaging. Recent advancements have demonstrated the capabilities of transient absorption microscopy (TAM) for label-free quantification of hemoglobin and stimulated Raman scattering (SRS) microscopy for pathological assessment of label-free virtual histochemical staining. We propose the combination of TAM and SRS with two-photon excited fluorescence (TPEF) to characterize, quantify, and compare hemodynamics, vessel structure, cell density, and cell identity in vivo between age groups.

Tissue imaging depth limit of stimulated Raman scattering microscopy.

Stimulated Raman scattering (SRS) microscopy is a promising technique for studying tissue structure, physiology, and function. Similar to other nonlinear optical imaging techniques, SRS is severely limited in imaging depth due to the turbidity and heterogeneity of tissue, regardless of whether imaging in the transmissive or epi mode. While this challenge is well known, important imaging parameters (namely maximum imaging depth and imaging signal to noise ratio) have rarely been reported in the literature.

Intraoperative assessment of skull base tumors using stimulated Raman scattering microscopy.

Intraoperative consultations, used to guide tumor resection, can present histopathological findings that are challenging to interpret due to artefacts from tissue cryosectioning and conventional staining. Stimulated Raman histology (SRH), a label-free imaging technique for unprocessed biospecimens, has demonstrated promise in a limited subset of tumors. Here, we target unexplored skull base tumors using a fast simultaneous two-channel stimulated Raman scattering (SRS) imaging technique and a new pseudo-hematoxylin and eosin (H&E) recoloring methodology.

Frequency Modulation Stimulated Raman Scattering Microscopy through Polarization Encoding.

Stimulated Raman scattering (SRS) microscopy is a powerful method for imaging molecular distributions based on their intrinsic vibrational contrast. However, despite a growing list of biological applications, SRS is frequently hindered by a parasitic background signal which both overpowers the signal in low-signal applications and makes the extraction of quantitative information from images challenging. Frequency modulation (FM) has been used to suppress this parasitic background.

Denoising of stimulated Raman scattering microscopy images via deep learning.

Stimulated Raman scattering (SRS) microscopy is a label-free quantitative chemical imaging technique that has demonstrated great utility in biomedical imaging applications ranging from real-time stain-free histopathology to live animal imaging. However, similar to many other nonlinear optical imaging techniques, SRS images often suffer from low signal to noise ratio (SNR) due to absorption and scattering of light in tissue as well as the limitation in applicable power to minimize photodamage. We present the use of a deep learning algorithm to significantly improve the SNR of SRS images.

Cellular Imaging Using Stimulated Raman Scattering Microscopy.

Cellular imaging is an active area of research that enables researchers to monitor cellular dynamics, as well as responses to various external stimuli (physiological stress, exogenous compounds, etc.). Stimulated Raman scattering (SRS) microscopy is one popular experimental tool used to image cells, largely because of its chemical specificity, high spatial resolution, and high image acquisition speed.

In Vitro Quantification of Single Red Blood Cell Oxygen Saturation by Femtosecond Transient Absorption Microscopy.

Hemoglobin, the oxygen carrying protein, ferries nearly all bodily oxygen from the lungs to cells and tissues in need. Blood oxygen saturation (sO) thus plays an important role in maintaining energy homeostasis throughout the body. Clinical and research tools have been developed to monitor sO at a wide range of temporal and spatial scales.

Perovskite Carrier Transport: Disentangling the Impacts of Effective Mass and Scattering Time Through Microscopic Optical Detection.

While carrier mobility is a practical and commonly cited measure of transport, it conflates the effects of two more fundamental material properties: the effective mass and mean scattering time of charge carriers. This Letter describes the correlation of two ultrafast imaging techniques to disentangle the effect of each on carrier transport in lead halide perovskites. Two materials are compared: methylammonium lead tri-iodide (MAPbI) and cesium lead bromide diiodide (CsPbBrI).

Screened Charge Carrier Transport in Methylammonium Lead Iodide Perovskite Thin Films.

While organometal halide perovskites are promising for a variety of optoelectronic applications, the morphological and compositional defects introduced by solution processing techniques have hindered efforts at understanding their fundamental properties. To provide a detailed picture of the intrinsic carrier transport properties of methylammonium lead iodide without contributions from defects such as grain boundaries, we utilized pump-probe microscopy to measure diffusion in individual crystalline domains of a thin film. Direct imaging of carrier transport in 25 individual domains yields diffusivities between 0.

Imaging theory of structured pump-probe microscopy.

With sub-micron spatial resolution and femtosecond temporal resolution, pump probe microscopy provides a powerful spectroscopic probe of complex electronic environments in bulk and nanoscale materials. However, the electronic structure of many materials systems are governed by compositional and morphological heterogeneities on length scales that lie below the diffraction limit. We have recently demonstrated Structured Pump Probe Microscopy (SPPM), which employs a patterned pump excitation field to provide spectroscopic interrogation of sub-diffraction limited sample volumes.