Note: Brillouin spectral analysis requires a single-longitudinal mode laser (~10 mW at the sample). For aligning purposes, use a strongly attenuated portion of this laser beam (<0.1 mW).
1. Initial Setup of Fiber and the EMCCD (Electron Multiplied Charge Coupled Device) Camera
2. Horizontal Stage of Spectrometer
3. Vertical Stage of Spectrometer
4. Combination of the Two Stages and Final Alignment
5. Measuring the Brillouin Shift
6. Calibration and Analysis of Brillouin Spectrum
Figure 2. Spectrometer calibration. (A) EMCCD camera frame obtained from calibration sample. (B) Lorentzian curve fit (red) to the measured data (blue). Please click here to view a larger version of this figure.
Figure 3 shows representative Brillouin spectra and their fits for different materials. The VIPAs both have a thickness of 5 mm which results in a FSR of approximately 20 GHz. The integration time for these measurements was 100 msec. 100 measurements were taken and averaged. One calibration measurement was taken prior to acquiring the spectra.
Figure 3. Brillouin Spectra of different materials. Lorentzian curve fit (red) to the measured data (blue). (a) Brillouin spectrum of Methanol. The measured Brillouin shift is 5.59 GHz. (b) Brillouin spectrum of Ethanol. The measured Brillouin shift is 5.85 GHz. (c) Brillouin spectrum of Polystyrene. The measured Brillouin shift is 14.12 GHz. Please click here to view a larger version of this figure.
The obtained Brillouin shifts agree with previously published data3,6,7. To determine if the alignment of the spectrometer is optimal, many spectral measurements of the same material can be taken sequentially, and the standard deviation of the peak positions can be calculated. Figure 4A shows a time-trace of 250 Brillouin measurements of methanol taken sequentially; a histogram of the evaluated Brillouin shifts is shown in Figure 4B. A well-aligned spectrometer with 5 mW of light at the sample and an integration time of 100 msec will have a standard deviation of σ ~ 10 MHz. Changes in Brillouin shift within corneal and lens tissue have been measured to be on the order of 1 GHz9,10,11. Therefore, Brillouin shift readings with variability of ≤10 MHz will enable the measurement of relevant mechanical variations in tissue.
Figure 4. Deviation in Brillouin shift over 250 methanol measurements. (A) Time-trace of 250 Brillouin measurements of methanol. (B) Histogram of Brillouin shift deviation over 250 measurements. Please click here to view a larger version of this figure.
OPTICS: | |||
VIPA (virtual image phase array) | LIGH MACHINERY | Quantity: 2 | |
Bundle of Three 423 Linear Stages with SM-25 Micrometers | NEWPORT | 423-MIC | Quantity: 1 |
SS Crossed-Roller Bearing Translation Stage, 0.5 in., 8-32, 1/4-20 | NEWPORT | 9066-X | Quantity: 1 |
Vernier Micrometer, 13 mm Travel, 9 lb Load Capacity, 50.8 TPI | NEWPORT | SM-13 | Quantity: 1 |
Adjustable Width Slit | NEWPORT | SV-0.5 | Quantity: 2 |
Compact Dovetail Linear Stage, 0.20 in. Z Travel, 1.57×1.57×1.38 in. | NEWPORT | DS40-Z | Quantity: 2 |
Slotted Base Plate, 25 or 40mm to 65mm Stage, 1.1 in. Range | NEWPORT | B-2B | Quantity: 2 |
Ø1/2" Optical Post, 8-32 Setscrew, 1/4"-20 Tap, L = 2", 5 Pack | THORLABS | TR2-P5 | Quantity: 2 |
Ø1/2" Post Holders, Spring-Loaded Hex-Locking Thumbscrews, L = 2", 5 Pack | THORLABS | PH2-P5 | Quantity: 1 |
Ø1/2" Post Holders, Spring-Loaded Hex-Locking Thumbscrew, L = 3", 5 Pack | THORLABS | PH3-P5 | Quantity: 1 |
Imperial Lens Mount For 2" Optics, 8-32 Tap | THORLABS | LMR2 | Quantity: 2 |
f=200.0 mm, Ø2" Achromatic Doublet, ARC: 400-700 nm | THORLABS | AC254-200-A | Quantity: 2 |
Kinematic Mount for up to 1.3" (33 mm) Tall Rectangular Optics, Right Handed | THORLABS | KM100C | Quantity: 2 |
Fixed Cylindrical Lens Mount, Max Optic Height: 1.60" (40.6 mm) | THORLABS | CH1A | Quantity: 2 |
f = 200.00 mm, H = 30.00 mm, L = 32.0 mm, N-BK7 Plano-Convex Cylindrical Lens, Antireflection Coating: 350-700 nm | THORLABS | L1653L1-A | Quantity: 2 |
Right-Angle Post Clamp, Fixed 90° Adapter | THORLABS | RA90 | Quantity: 1 |
Adapter with External C-Mount Threads and Internal SM1 Threads | THORLABS | SM1A9 | Quantity: 1 |
Studded Pedestal Base Adapter, 1/4"-20 Thread | THORLABS | PB4 | Quantity: 2 |
Spacer, 2" x 3", 1.000" Thick | THORLABS | Ba2S7 | Quantity: 2 |
543 nm, f=15.01 mm, NA=0.17 FC/APC Fiber Collimation Pkg. | THORLABS | F260APC-A | Quantity: 1 |
SM1-Threaded Adapter for Ø11 mm collimators | THORLABS | Ad11F | Quantity: 1 |
Translating Lens Mount for Ø1" Optics, 1 Retaining Ring Included | THORLABS | LM1XY | Quantity: 1 |
Single Mode Patch Cable, 450 – 600 nm, FC/APC, 2 m Long | THORLABS | P3-460B-FC-2 | Quantity: 1 |
1:1 Matched Achr. Pair, f1=30 mm, f2=30 mm, BBAR 400-700 nm | THORLABS | MAP103030-A | Quantity: 1 |
SM1 Lens Tube…length to adjust depend on CCD, we have 3.5 inches | THORLABS | SM1LXX | Quantity: 1 |
Base Adapters for Ø1/2" Post Holders and Ø1" Posts | THORLABS | BE1 | Quantity: 8 |
Clamping Forks for Ø1/2" Post Holders and Ø1" Posts | THORLABS | CF125 | Quantity: 8 |
HW-KIT5 – 4-40 Cap Screw and Hardware Kit for Mini-Series | THORLABS | HW-KIT5 | Quantity: 1 |
D20S – Standard Iris, Ø20.0 mm Max Aperture | THORLABS | D20S | Quantity: 2 |
FOR ENCLOSURE | |||
25 mm Construction Rail, L = 21" | THORLABS | XE25L21 | Quantity: 6 |
1" Construction Cube with Three 1/4" (M6) Counterbored Holes | THORLABS | RM1G | Quantity: 8 |
Right-Angle Bracket for 25 mm Rails | THORLABS | XE25A90 | Quantity: 12 |
25 mm Construction Rail, L = 15" | THORLABS | XE25L15 | Quantity: 4 |
25 mm Construction Rail, L = 9" | THORLABS | XE25L09 | Quantity: 8 |
High Performance Black Masking Tape, 2" x 60 yds. (50 mm x 55 m) Roll | THORLABS | T743-2.0 | Quantity: 1 |
Low-Profile T-Nut, 1/4"-20 Tapped Hole, Qty: 10 | THORLABS | XE25T3 | Quantity: 1 |
1/4"-20 Low-Profile Channel Screws (100 Screws/Box) | THORLABS | SH25LP38 | Quantity: 1 |
60" (W) x 3 yds. (L) x 0.005" (T) (1.5 m x 2.7 m x 0.12 mm) Blackout Fabric | THORLABS | BK5 | Quantity: 1 |
CAMERA, LASER and MICROSCOPE | |||
EMCCD camera | ANDOR | iXon Ultra 897 | Quantity: 1 |
400 mW single mode green laser | LASER QUANTUM | torus 532 | Quantity: 1 |
Research Inverted System Microscope | OLYMPUS | IX71 | Quantity: 1 |
The goal of this protocol is to build a parallel high-extinction and high-resolution optical Brillouin spectrometer. Brillouin spectroscopy is a non-contact measurement method that can be used to obtain direct readouts of viscoelastic material properties. It has been a useful tool in material characterization, structural monitoring and environmental sensing. In the past, Brillouin spectroscopy has usually employed scanning Fabry-Perot etalons to perform spectral analysis. This process requires high illumination power and long acquisition times, making the technique unsuitable for biomedical applications. A recently introduced novel spectrometer overcomes this challenge by employing two VIPAs in a cross-axis configuration. This innovation enables sub-Gigahertz (GHz) resolution spectral analysis with sub-second acquisition time and illumination power within the safety limits of biological tissue. The multiple new applications facilitated by this improvement are currently being explored in biological research and clinical application.
The goal of this protocol is to build a parallel high-extinction and high-resolution optical Brillouin spectrometer. Brillouin spectroscopy is a non-contact measurement method that can be used to obtain direct readouts of viscoelastic material properties. It has been a useful tool in material characterization, structural monitoring and environmental sensing. In the past, Brillouin spectroscopy has usually employed scanning Fabry-Perot etalons to perform spectral analysis. This process requires high illumination power and long acquisition times, making the technique unsuitable for biomedical applications. A recently introduced novel spectrometer overcomes this challenge by employing two VIPAs in a cross-axis configuration. This innovation enables sub-Gigahertz (GHz) resolution spectral analysis with sub-second acquisition time and illumination power within the safety limits of biological tissue. The multiple new applications facilitated by this improvement are currently being explored in biological research and clinical application.
The goal of this protocol is to build a parallel high-extinction and high-resolution optical Brillouin spectrometer. Brillouin spectroscopy is a non-contact measurement method that can be used to obtain direct readouts of viscoelastic material properties. It has been a useful tool in material characterization, structural monitoring and environmental sensing. In the past, Brillouin spectroscopy has usually employed scanning Fabry-Perot etalons to perform spectral analysis. This process requires high illumination power and long acquisition times, making the technique unsuitable for biomedical applications. A recently introduced novel spectrometer overcomes this challenge by employing two VIPAs in a cross-axis configuration. This innovation enables sub-Gigahertz (GHz) resolution spectral analysis with sub-second acquisition time and illumination power within the safety limits of biological tissue. The multiple new applications facilitated by this improvement are currently being explored in biological research and clinical application.