| dc.contributor.advisor | Mondal, Partha P | |
| dc.contributor.author | Basumatary, Jigmi | |
| dc.date.accessioned | 2020-07-13T07:03:15Z | |
| dc.date.available | 2020-07-13T07:03:15Z | |
| dc.date.submitted | 2019 | |
| dc.identifier.uri | https://etd.iisc.ac.in/handle/2005/4490 | |
| dc.description.abstract | Optical fluorescence microscopy is one of the key tool, that is indispensable for the study
of complex and dynamic biological processes. Key issues associated with the existing fluorescence
imaging systems are, photo-bleaching, long-time imaging and low-quality volume
reconstruction. Continuous monitoring of large specimens for long duration requires fast volume
imaging. This is essential for understanding processes occurring during developmental
stages of multi-cellular organisms. One of the key obstacles for prolonged monitoring and data
collection is photo-bleaching. To overcome the effect of bleaching, a single and multi- color
Lightsheet Illuminated Volume Expedite (LIVE) imaging technique is developed that enables
rapid screening of multiple tissues in an organism with an order-less photo-bleaching. Our approach
based on LIVE imaging employs quantized step rotation of the specimen to record 2D
angular data that reduces data collection time by a factor of more than 10 when compared to
existing light sheet fluorescence microscopy. A co-planar multicolor light sheet is generated
to excite spectrally-separated fluorescent probes that label the target tissues. Arduino-based
control systems were employed to automatize and control the volume data acquisition process.
To illustrate the advantages of our approach, we have noninvasively imaged Drosophila larvae
(without removing chitinous cover) and Zebrafish embryo. Dynamic studies of multiple organs
(muscle and yolk-sac) in Zebrafish over a prolonged duration of time (5 days) were carried out
to understand muscle structuring and metabolism. Volume reconstruction, intensity plots and
inter-dependence ratio analysis allowed us detailed visualization of organs during early development
(Pharyngula period) in Zebrafish. The advantage of multicolor lightsheet illumination, fast
volume imaging, simultaneous multiple organ imaging and order-less photo-bleaching makes
LIVE imaging the system of choice for rapid monitoring and real-time assessment of macroscopic
biological organisms at microscopic resolutions. We envision developed LIVE system
as the next generation state-of-art-the-art light sheet fluorescence imaging | en_US |
| dc.language.iso | en_US | en_US |
| dc.rights | I grant Indian Institute of Science the right to archive and to make available my thesis or dissertation in whole or in part in all forms of media, now hereafter known. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part
of this thesis or dissertation | en_US |
| dc.subject | fluorescence imaging systems | en_US |
| dc.subject.classification | Research Subject Categories::TECHNOLOGY::Information technology::Image analysis | en_US |
| dc.title | Rapid Lightsheet Volume Imaging For Developmental Biology | en_US |
| dc.type | Thesis | en_US |
| dc.degree.name | MTech (Res) | en_US |
| dc.degree.level | Masters | en_US |
| dc.degree.grantor | Indian Institute of Science | en_US |
| dc.degree.discipline | Engineering | en_US |
