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Fluorescence based rapid optical volume screening system (OVSS) for interrogating multicellular organisms.

Jigmi Basumatary1, Tarannum Ara1, Amartya Mukherjee2

  • 1Nanobioimaging Lab., Department of Instrumentation and Applied Physics, Indian Institute of Science, Bangalore, 560012, India.

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|April 8, 2021
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Summary
This summary is machine-generated.

We developed a fast, multicolor lightsheet imaging technique (OVSS) to monitor large biological specimens over time, overcoming photobleaching. This method enables rapid screening of multiple tissues and dynamic studies of developmental processes in organisms like Zebrafish.

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Area of Science:

  • Developmental Biology
  • Microscopy Techniques
  • Bioimaging

Background:

  • Long-term monitoring of large specimens requires fast volume imaging for developmental studies.
  • Photobleaching is a major limitation in fluorescence-based prolonged monitoring.
  • Existing light sheet microscopy (SPIM) can be slow for rapid screening.

Purpose of the Study:

  • To develop a novel imaging technique for rapid, prolonged monitoring of large biological specimens.
  • To overcome photobleaching limitations in fluorescence microscopy.
  • To enable simultaneous visualization of multiple tissues and dynamic processes.

Main Methods:

  • Developed single- and multi-color Orthogonal Volume Scanning System (OVSS) imaging.
  • Employed quantized step rotation for reduced data acquisition time compared to SPIM.
  • Utilized a co-planar multicolor light sheet and a dual-channel detection sub-system.
  • Automated data acquisition using Arduino-based control systems.

Main Results:

  • Successfully imaged Drosophila larvae and Zebrafish embryos noninvasively.
  • Conducted dynamic studies of Zebrafish organs (muscle, yolk-sac) for 5 days.
  • Analyzed muscle structuring, primitive macrophages, and lipid-to-protein metabolism transition.
  • Demonstrated OVSS imaging's ability to monitor metabolic shifts during Zebrafish development.

Conclusions:

  • OVSS imaging provides fast volume scanning and simultaneous multi-organ visualization.
  • The technique minimizes photobleaching, making it ideal for prolonged studies.
  • OVSS imaging offers microscopic resolution for macroscopic biological organisms, facilitating real-time assessment.