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Abstract Slow acquisition time and instrument stability are the two major limitations for the application of confocal Brillouin microscopy to biological materials. Although overlooked, coupling the microscope to the spectrometer with a multimode fiber (MMF) is a simple yet viable solution to increase both the detection efficiency and the stability of the classical single-mode fiber-coupled virtually imaged phase array (VIPA) instruments. Here we implement the first successful MMF-coupled VIPA spectrometer for confocal Brillouin applications and present a dimensioning strategy to optimize its collected power. The use of an MMF brings a tremendous improvement on the stability of the spectrometer that allows performing experiments over several weeks without realignment of the device. For instance, we map the Brillouin shift and linewidth in growing ductal and acinar organoids with a spatial resolution of 1 × 1 × 6 μ m 3 and 50 ms dwell time. Our results clearly reveal the formation of a lumen in these organoids. Careful examination of the data also suggests an increase in the viscosity of the cells of the assembly.
Bouvet et al. (Mon,) studied this question.