Scattering oblique plane microscopy for in-vivo blood cell imaging

TL;DR

This paper introduces a scattering-contrast oblique plane microscope (sOPM) for label-free, in vivo blood cell imaging in humans, enabling high-resolution, volumetric visualization of blood flow without exogenous dyes.

Contribution

The study develops and demonstrates a novel scattering-contrast OPM technique for in vivo human blood cell imaging, expanding OPM applications beyond animal models and fluorescence imaging.

Findings

Successful label-free imaging of blood cells in vivo

Differentiation of cellular and acellular absorption gaps

Comparison of fluorescence and scattering OPM techniques

Abstract

Oblique plane microscopy (OPM) enables high speed, volumetric fluorescence imaging through a single-objective geometry. While these advantages have positioned OPM as a valuable tool to probe biological questions in animal models, its potential for in vivo human imaging is largely unexplored due to its typical use with exogenous fluorescent dyes. Here we introduce a scattering-contrast oblique plane microscope (sOPM) and demonstrate label-free imaging of blood cells flowing through human capillaries in vivo. The sOPM illuminates a capillary bed in the ventral tongue with an oblique light sheet, and images side- and back- scattered signal from blood cells. By synchronizing sOPM with a conventional capillaroscope, we acquire paired widefield and axial images of blood cells flowing through a capillary loop. The widefield capillaroscope image provides absorption contrast and confirms the presence of red blood cells (RBCs), while the sOPM image may aid in determining whether optical absorption gaps (OAGs) between RBCs have cellular or acellular composition. Further, we demonstrate consequential differences between fluorescence and scattering versions of OPM by imaging the same polystyrene beads sequentially with each technique. Lastly, we substantiate in vivo observations by imaging isolated red blood cells, white blood cells, and platelets in vitro using 3D agar phantoms. These results demonstrate a promising new avenue towards in vivo blood analysis.