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Light sheet based volume flow cytometry (VFC) for rapid volume reconstruction and parameter estimation on the go

Kumar, P and Joshi, P and Basumatary, J and Mondal, PP (2022) Light sheet based volume flow cytometry (VFC) for rapid volume reconstruction and parameter estimation on the go. In: Scientific Reports, 12 (1).

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Official URL: https://doi.org/10.1038/s41598-021-03902-8


Optical imaging is paramount for disease diagnosis and to access its progression over time. The proposed optical flow imaging (VFC/iLIFE) is a powerful technique that adds new capabilities (3D volume visualization, organelle-level resolution, and multi-organelle screening) to the existing system. Unlike state-of-the-art point-illumination-based biomedical imaging techniques, the sheet-based VFC technique is capable of single-shot sectional visualization, high throughput interrogation, real-time parameter estimation, and instant volume reconstruction with organelle-level resolution of live specimens. The specimen flow system was realized on a multichannel (Y-type) microfluidic chip that enables visualization of organelle distribution in several cells in-parallel at a relatively high flow-rate (2000 nl/min). The calibration of VFC system requires the study of point emitters (fluorescent beads) at physiologically relevant flow-rates (500�2000 nl/min) for determining flow-induced optical aberration in the system point spread function (PSF). Subsequently, the recorded raw images and volumes were computationally deconvolved with flow-variant PSF to reconstruct the cell volume. High throughput investigation of the mitochondrial network in HeLa cancer cell was carried out at sub-cellular resolution in real-time and critical parameters (mitochondria count and size distribution, morphology, entropy, and cell strain statistics) were determined on-the-go. These parameters determine the physiological state of cells, and the changes over-time, revealing the metastatic progression of diseases. Overall, the developed VFC system enables real-time monitoring of sub-cellular organelle organization at a high-throughput with high-content capacity. © 2022, The Author(s).

Item Type: Journal Article
Publication: Scientific Reports
Publisher: Nature Research
Additional Information: The copyright for this article belongs to Authors
Department/Centre: Division of Physical & Mathematical Sciences > Instrumentation Appiled Physics
Date Deposited: 27 Jan 2022 11:43
Last Modified: 27 Jan 2022 11:43
URI: http://eprints.iisc.ac.in/id/eprint/71008

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