Curtailed light sheet microscopy for rapid imaging of macroscopic biological specimens |
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Authors: | Chelur K. Rasmi Mani Madhangi Upendra Nongthomba Partha Pratim Mondal |
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Affiliation: | 1. Nanobioimaging Laboratory, Department of Instrumentation and Applied Physics, Indian Institute of Science, Bangalore, Karnataka, India;2. Department of Physics, Indian Institute of Science, Bangalore, Karnataka, India;3. Department of Molecular Reproduction and Development Group, Indian Institute of Science, Bangalore, Karnataka, India |
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Abstract: | We study the feasibility of volume imaging from a few angular views/scans in a light sheet fluorescence microscopy. Two‐dimensional (2D) images (angular views) were acquired at an angular separation of 10° and volume images were constructed with merely 18, 9, and 6 views. We study the structural changes in a 5‐day old Zebrafish embryo labeled with Phalloidin TRITC that binds to F‐Actin of embryo cell. To collect the data, the specimen is rotated (for varying sampling angles Δθ) with respect to a fixed vertical axis passing through the volume‐of‐interest (yolk sac). In the proposed realization of selective plane illumination microscopy (SPIM) technique, the translation is completely avoided. Analysis shows rich structural information with marginal reduction in contrast. Comparison with the state‐of‐the‐art SPIM shows appreciable volume reconstruction (from an order less 2D scans) that may be good enough for rapid monitoring of macroscopic specimens. Microsc. Res. Tech. 79:455–458, 2016. © 2016 Wiley Periodicals, Inc. |
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Keywords: | fluorescence microscopy light sheet microscopy biological imaging |
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