研究目的
To directly measure cellular forces and mechanical properties in vivo during embryogenesis using a setup of optical tweezers coupled to a light sheet microscope.
研究成果
Optical tweezers allow for non-invasive, absolute mechanical measurements directly in the developing embryonic epithelium, presenting advantages over other methods. The technique provides insights into cell mechanics and tissue morphogenesis, with potential for further improvements and applications.
研究不足
The method relies on the measurements of the cell contacts' position, requiring a high-contrast fluorescent marker. The quality of the objective lens and the laser beam are critical to effective trapping. The calibration process for absolute measurements can be tedious, and the technique may not be versatile enough for all tissues due to higher tension or difficult access.
1：Experimental Design and Method Selection:
The setup involves optical tweezers coupled to a light sheet microscope to apply controlled forces on cell-cell contacts in the early Drosophila embryo.
2：Sample Selection and Data Sources:
Drosophila embryos are used as samples, with specific stages selected for experiments.
3：List of Experimental Equipment and Materials:
Includes a 1070 nm laser unit, optical shutter, galvanometer mirrors, fluorescent beads, and a light sheet microscope.
4：Experimental Procedures and Operational Workflow:
Detailed steps for setting up the optical tweezers and light sheet microscope, calibrating the trap position with beads, mounting Drosophila embryos, and performing trapping experiments in vivo.
5：Data Analysis Methods:
Analysis involves generating kymographs, detecting interface positions with subpixel resolution, and performing mechanical measurements such as stiffness and tension measurements.
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