研究目的
Investigating the use of light droplet illumination to suppress high-energy side lobes in Bessel beams for enhanced contrast in light sheet fluorescence microscopy.
研究成果
The light droplet illumination demonstrated a significant reduction in the side lobes of Bessel beams, leading to enhanced imaging contrast in light sheet fluorescence microscopy. This method offers a promising approach for rapid, high-contrast volume imaging of biological specimens.
研究不足
The study focuses on the suppression of side lobes in Bessel beams for light sheet microscopy but does not explore the potential for further optimization in terms of depth of focus or application in nonlinear regimes.
1:Experimental Design and Method Selection:
The study involved generating a light droplet illumination by interfering Bessel beams of specific k-vectors to suppress side lobes. The setup included a spatial light modulator (SLM) for beam modulation and objective lenses for Fourier transform and fluorescence signal collection.
2:Sample Selection and Data Sources:
Fluorescent spheres embedded in a fixed gel were used as the sample to demonstrate the imaging contrast enhancement.
3:List of Experimental Equipment and Materials:
The setup included an SLM (Holoeye LC2012), objective lenses (Olympus Plan N 10X, Zeiss LD A-Plan 20x Ph1 M27), a tube lens (Thorlabs TTL200), a bandpass filter (Thorlabs FELH0550), and a CCD camera (Photometrics CoolSNAP HQ2).
4:2). Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: The wavefront of a laser beam was modulated by the SLM to generate the droplet illumination. The fluorescent signal from the sample was collected and detected to compare the imaging contrast between standard Bessel beam and droplet illuminations.
5:Data Analysis Methods:
The optical power and FWHM of the droplet side lobes were measured and compared with those of a Bessel beam. The imaging contrast was evaluated based on the fluorescence images obtained.
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objective lens
Plan N 10X
Olympus
Performing a Fourier transform of the modulated annular fields and generating the light droplet illumination
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objective lens
LD A-Plan 20x Ph1 M27
Zeiss
Collecting the fluorescent signal from the sample
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tube lens
TTL200
Thorlabs
Assisting in the collection of the fluorescent signal
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bandpass filter
FELH0550
Thorlabs
Selecting only the fluorescent signal
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spatial light modulator
LC2012
Holoeye
Modulating the wavefront of an expanded collimated laser beam
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CCD camera
CoolSNAP HQ2
Photometrics
Detecting the fluorescent signal
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