研究目的
To develop a versatile high-speed confocal microscopy system using a single laser beam for improved imaging quality and reduced phototoxicity, and to extend its functionality with an astigmatism particle tracking velocimetry (APTV) device for three-dimensional dynamics measurement.
研究成果
The developed confocal microscopy system using a single laser beam achieves high-speed imaging with reduced phototoxicity and no crosstalk, maintaining high resolution independent of frame rate. The integration with APTV allows for simultaneous high-resolution confocal imaging and three-dimensional dynamics measurement, offering new possibilities for research in colloidal and granular systems, as well as wetting phenomena.
研究不足
The resolution depends on the quality of the sample, with ideal conditions yielding a lateral resolution of 0.235 μm but practical samples showing reduced resolution. The axial resolution is inherently lower than lateral, and the system's performance is limited by the scanning units' speed and the camera's sensor size and pixel size.
1:Experimental Design and Method Selection:
The setup includes a single laser beam confocal microscope with a re-scan configuration using a second 2D scan unit to project images directly onto a 2D CCD-sensor. This design eliminates crosstalk and maintains imaging quality independent of frame rate.
2:Sample Selection and Data Sources:
The system is tested with reflective calibration patterns and dense colloidal probes to evaluate resolution and imaging quality.
3:List of Experimental Equipment and Materials:
Includes a green laser light source (Cobolt Samba 532 nm, 100 mW), resonant and galvanic scanners, telecentric system, piezo-positioning system, and a high-speed camera.
4:Experimental Procedures and Operational Workflow:
The system operates at high frame rates up to 1 kHz for xy-scans and 200 Hz for xz-scans, with a z-amplitude of 100 μm. The setup allows for flexible scanning modes including xy, xz, and xyz volumes.
5:Data Analysis Methods:
Resolution is determined by analyzing reflective calibration grids and colloidal samples, with edge steepness and line width used as indicators for lateral and axial resolution.
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