Holographic optical tweezers (HOTs) extend optical trapping into three dimensions. Volume imaging then becomes a concern as trapped objects are easily moved out of focus of the imaging objective lens. Here we demonstrate a three-dimensional real-time interactive optical trapping, manipulating, and imaging system based on HOTs incorporated with volume holographic microscope. Intensity information about the trapped objects at multiple depths can be captured in a single measurement. This method is compatible with most imaging modes such as bright-field and fluorescence.
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