Volumetric chemical imaging in vivo by a remote-focusing stimulated Raman scattering microscope

Abstract

Operable under ambient light and providing chemical selectivity, stimulated Raman scattering (SRS) microscopy opens a new window for imaging molecular events on a human subject, such as filtration of topical drugs through the skin. A typical approach for volumetric SRS imaging is through piezo scanning of an objective lens, which often disturbs the sample and offers a low axial scan rate. To address these challenges, we have developed a deformable mirror-based remote-focusing SRS microscope, which not only enables high-quality volumetric chemical imaging without mechanical scanning of the objective but also corrects the system aberrations simultaneously. Using the remote-focusing SRS microscope, we performed volumetric chemical imaging of living cells and captured in real time the dynamic diffusion of topical chemicals into human sweat pores.

Investigation of the Spatial Generation of Stimulated Raman Scattering Using Computer Simulation and Experimentation

“…The difference is mainly caused by the filtering of the experimental data and the imaging setup, but part of the difference may be explained by the difference in the original beam profile. The simulations also showed that the position of the maximum intensity in the Stokes gain comes after the maximum intensity of the pump beam, which confirms the theoretical prediction made by Carman et al 19…”

“…SRS microscopy is a commonly used imaging technique where the SRS image is constructed by point scanning the sample using a confocal microscope. 10 This type of SRS microscopy has been used for live cell imaging, [11][12][13] imaging of cancerous tissue, [14][15][16] and volumetric 3D imaging [17][18][19][20] . Modifications can be made to the experimental setup to allow for probing of several Raman shifts simultaneously, this is usually called multiplex SRS (MSRS).…”

Diagnostics and Therapy Assessment Using Label-Free Raman Imaging

“…171 In addition, Lin et al constructed a deformable mirror-based remote-focusing SRS system, enabling excellent volumetric imaging of the skin (Figure 8). 172 In this study, the authors obtained the dynamic diffusion of drugs in real-time into human sweat pores, which were suggested to disrupt the stratum corneum barrier and deliver the drugs into the epidermal layer. 172 Cell-Based Therapy.…”

“…172 In this study, the authors obtained the dynamic diffusion of drugs in real-time into human sweat pores, which were suggested to disrupt the stratum corneum barrier and deliver the drugs into the epidermal layer. 172 Cell-Based Therapy. Cell-based therapy is considered a strategy for the treatment of many human diseases.…”

Adaptive Optical Coherent Raman Imaging of Axons through Mouse Cranial Bone

“…[19][20][21] With its simplicity of implementation, sensorless AO has been employed in CRS microscopy to compensate for sample-and system-induced aberrations. 22,23 Still, this AO technique has a limitation that aberration should be mild enough for the sample to be initially visible for evaluating quality metrics, meaning that the imaging depth increase is marginal. The other approach is wavefront sensing AO, which directly measures the wavefront of returning waves to determine the sample-induced aberrations.…”