Pulsed heterodyne detection enables fiber-compatible, high-specificity Brillouin biomechanics in intact tissue and the living brain
Brillouin microscopy provides label-free, three-dimensional mechanical characterization of biological specimens, but current dispersive detection imposes two limits: the fiber background folds onto the sample spectrum, precluding single-fiber operation, and a ~250 MHz dispersion-induced instrumental broadening blurs mechanically distinct components within a focal volume. Here, we introduce pulsed heterodyne Brillouin detection (PHBD), which retrieves the spectrum electronically from temporal beat notes, overcoming both limitations. The fiber background beats outside the detection band and is rejected; elimination of dispersive broadening yields 25-MHz spectrometer resolution. Pulsed excitation reaches Brillouin-signal-shot-noise-limited detection, attaining 9.0-MHz shift precision in 3 ms at 30 mW in water with 53-fold improvement in energy efficiency over continuous-wave excitation. Through a bare 125-um fiber, PHBD resolves regional contrast along a 4-mm insertion track in the living mouse brain; in free space, it resolves distinct Brillouin components from the cell wall and adjacent cytoplasm in strongly scattering Arabidopsis root tips with epi-mode.