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Spontaneous Brillouin spectroscopy has been established, in recent years, as a valuable approach for the mechanical analysis of soft materials, such as liquids, real tissue, tissue phantoms and biological cells1,2,3,4,5,6,7. In this approach, a single laser illuminates the sample and light that is inelastically scattered from spontaneous thermal acoustic waves in the medium is collected by a spectrometer, providing useful information on the viscoelastic properties of the sample. The spontaneous Brillouin spectrum includes two Brillouin peaks at the acoustic Stokes and anti-Stokes resonances of the material, and a Rayleigh peak at the illuminating laser frequency (due to elastically scattered light). For a Brillouin backscattering geometry, the Brillouin frequencies are shifted by several GHz from the illuminating laser frequency and have spectral width of hundreds of MHz.
While scanning Fabry-Perot spectrometers have been the systems-of-choice for acquiring spontaneous Brillouin spectra in soft matter1,2, recent technological advances in virtually imaged phase array (VIPA) spectrometers have enabled significantly faster (sub-second) Brillouin measurements with adequate spectral-resolution (sub-GHz)3,4,5,6,7. In this protocol, we present the construction of a different, high-speed, high spectral-resolution, accurate Brillouin spectrometer based on the detection of continuous-wave-stimulated-Brillouin-scattering (CW-SBS) light from non-turbid and turbid samples in a nearly back scattering geometry.
In CW-SBS spectroscopy, continuous-wave (CW) pump and probe lasers, slightly detuned in frequency, overlap in a sample to stimulate acoustic waves. When the frequency difference between the pump and probe beams matches a specific acoustic resonance of the material, amplification or deamplification of the probe signal is provided by stimulated Brillouin gain or loss (SBG/SBL) processes, respectively; otherwise, no SBS (de)amplification occurs8,9,10,11. Thus, an SBG (SBL) spectrum can be acquired by scanning the frequency difference between the lasers across the material Brillouin resonances and detecting the increase (decrease), or gain (loss), in the probe intensity due to SBS. Unlike in spontaneous Brillouin scattering, elastic scattering background is inherently absent in SBS, enabling excellent Brillouin contrast in both turbid and non-turbid samples without any need for Rayleigh rejection filters as required in VIPA spectrometers10,11,13.
The main building blocks of a CW-SBS spectrometer are the pump and probe lasers and the stimulated Brillouin gain/loss detector. For high spectral-resolution, high speed CW-SBS spectroscopy, the lasers need to be single-frequency (< 10 MHz linewidth) with sufficiently wide wavelength tunability (20 - 30 GHz) and scanning rate (> 200 GHz/s), long-term frequency stability (<50 MHz/h) and low intensity noise. Furthermore, linearly polarized and diffraction-limited laser beams with powers of few hundreds (tens) of mW on the sample are required for the pump (probe) beam. Finally, the stimulated Brillouin gain/loss detector should be designed to reliably detect weak backward stimulated Brillouin gain/loss (SBG/SBL) levels (10-5 - 10-6) in soft matter. To meet these needs, we selected distributed feedback (DFB) diode lasers coupled to polarization-maintaining fibers along with a stimulated Brillouin gain/loss detector combining an ultra-narrowband atomic vapor notch-filter and a high-frequency single-modulation lock-in amplifier as illustrated in Figure 1. This detection scheme doubles the intensity of the SBG signal while significantly reducing noise in the probe intensity, where the desired SBG signal is embedded11. Note that the role of the atomic vapor notch-filter used in our SBS spectrometer is to significantly reduce the detection of unwanted stray pump reflections rather than to decrease the elastic scattering background as in VIPA spectrometers that detect both spontaneous Rayleigh and Brillouin scattered light. Using the protocol detailed below, a CW-SBS spectrometer can be constructed with the capability of acquiring transmission spectra of water and tissue phantoms with SBG levels as low as 10-6 at <35 MHz Brillouin-shift measurement precision and within 100 ms or less.

Figure 1: Continuous-wave Stimulated Brillouin Scattering (CW-SBS) Spectrometer. Two continuous-wave pump and probe diode lasers (DL), frequency detuned around the Brillouin shift of the sample, are coupled into polarization-maintaining single-mode fibers with collimators C1 and C2, respectively. The pump-probe frequency difference is measured by detecting the beat frequency between beams peeled from the pump and probe lasers using a set of fiber splitters (FS), a fast photodetector (FPD), and a frequency counter (FC). The S-polarized probe beam (light red), expanded using a Keplerian beam expander (L1 and L2), is right circularly polarized by a quarter-wave plate (λ1/4) and focused on the sample (S) by an achromatic lens (L3). For effective SBS interaction and optical isolation, the pump beam (deep red), expanded using a Keplerian beam expander (L5 and L6), is first P-polarized using a half-wave plate λ2/4), then transmitted through a polarizing beam splitter (PBS), and is finally left circularly polarized by a quarter-wave plate (λ2/4) and focused on the sample with an achromatic lens (L4; same as L3). Note that the pump and probe beams nearly counter-propagate in the sample and that an S-oriented polarizer (P) was used to prevent the P-polarized pump beam (coming out of λ1/4) from entering the probe laser. For lock-in detection, the pump beam is sinusoidally modulated at fm with an acousto-optical modulator (AOM). The SBG signal, manifested as intensity variations at frequency fm (see inset), is demodulated with a lock-in amplifier (LIA) following detection by a large-area photodiode (PD). For significant elimination of stray pump reflections in the photodiode, a narrowband Bragg filter (BF) and an atomic notch filter (85RB) around the pump wavelength are used alongside with a light-blocking iris (I). Data is recorded by a data acquisition card (DAQ) connected to a personal computer (PC) for further analysis of the Brillouin spectrum. All folding mirrors (M1-M6) are used to fit the spectrometer on a 18''×24'' breadboard that is vertically mounted on the optical table for facilitating placement of watery samples. Please click here to view a larger version of this figure.