10 February 2011 Digital holographic microscopy combined with optical tweezers
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Abstract
While optical tweezers have been widely used for the manipulation and organization of microscopic objects in three dimensions, observing the manipulated objects along axial direction has been quite challenging. In order to visualize organization and orientation of objects along axial direction, we report development of a Digital holographic microscopy combined with optical tweezers. Digital holography is achieved by use of a modified Mach-Zehnder interferometer with digital recording of interference pattern of the reference and sample laser beams by use of a single CCD camera. In this method, quantitative phase information is retrieved dynamically with high temporal resolution, only limited by frame rate of the CCD. Digital focusing, phase-unwrapping as well as online analysis and display of the quantitative phase images was performed on a software developed on LabView platform. Since phase changes observed in DHOT is very sensitive to optical thickness of trapped volume, estimation of number of particles trapped in the axial direction as well as orientation of non-spherical objects could be achieved with high precision. Since in diseases such as malaria and diabetics, change in refractive index of red blood cells occurs, this system can be employed to map such disease-specific changes in biological samples upon immobilization with optical tweezers.
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Nelson Cardenas, Lingfeng Yu, Samarendra K. Mohanty, "Digital holographic microscopy combined with optical tweezers", Proc. SPIE 7902, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX, 79020U (10 February 2011); doi: 10.1117/12.875992; https://doi.org/10.1117/12.875992
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