研究目的
To integrate a spatial light modulator (SLM) into the optical path of an optical-resolution photoacoustic microscopy (OR-PAM) for realizing flexibly adjustable depth-of-focus (DoF) to acquire volumetric images of biological specimens with consistent lateral resolution.
研究成果
The integration of an SLM into the OR-PAM system allows for flexible adjustment of the DoF, enabling volumetric imaging with consistent lateral resolution. This capability is valuable for biomedical applications requiring imaging of specimens with varying thicknesses.
研究不足
The study demonstrates the technique with tungsten filaments and a hair as samples, indicating the need for further validation with biological specimens. The DoF in tissue-mimicking phantoms is reduced compared to water, suggesting limitations in highly scattering media.
1:Experimental Design and Method Selection:
The study integrates an SLM into the OR-PAM system to modulate the pupil mask of the optical objective, enabling flexible adjustment of the light fields in the focus by assigning variable phase patterns onto the SLM.
2:Sample Selection and Data Sources:
The method is demonstrated by imaging a phantom made from tungsten filaments positioned at different depths.
3:List of Experimental Equipment and Materials:
Includes a frequency-doubled Nd:YAG laser, lenses, polarizer, half-wave plate, reflective phase-only SLM, microscope objective, ultrasonic transducer, and a 3D scanning stage.
4:Experimental Procedures and Operational Workflow:
The laser output is collimated, polarized, and then modulated by the SLM to produce desired light field distributions in the focal region. PA signals are detected, amplified, and digitized for analysis.
5:Data Analysis Methods:
The lateral resolution and DoF are evaluated by imaging carbon filaments and analyzing the full-width-at-half-maximum (FWHM) values of the PA amplitudes.
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microscope objective
RMS4X
Olympus
Used to focus the modulated light onto the sample.
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3D scanning stage
PLS-85
PI
Used to mount and move the sample during imaging.
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power meter
S121C
Thorlabs
Used to measure the light transmittance.
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Nd:YAG laser
SPOT-532
Elforlight
Served as the PA excitation source.
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polarizer
LPVISA100-MP2
Thorlabs
Used to polarize the laser output.
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half-wave plate
WPH10M-532
Thorlabs
Used in combination with the polarizer to polarize the laser output.
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spatial light modulator
LETO-HES 4550
Holoeye
Used to modulate the pupil mask of the optical objective.
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ultrasonic transducer
Used to detect the PA signals.
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low-noise amplifier
ZFL-500LN-BNC+
Mini-Circuits
Used to amplify the detected PA signals.
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data acquisition card
ATS9325
Alazar
Used to digitize the amplified PA signals.
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interconnect card
PCIe-6321
National Instruments
Used to trigger PA laser firing and data acquisition, as well as to control raster scanning.
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