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Slanted channel microfluidic chip for 3D fluorescence imaging of cells in flow

Jagannadh, Veerendra Kalyan and Mackenzie, Mark D and Pal, Parama and Kar, Ajoy K and Gorthi, Sai Siva (2016) Slanted channel microfluidic chip for 3D fluorescence imaging of cells in flow. In: OPTICS EXPRESS, 24 (19). pp. 22144-22158.

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Official URL: http://dx.doi.org/10.1364/OE.24.022144

Abstract

Three-dimensional cellular imaging techniques have become indispensable tools in biological research and medical diagnostics. Conventional 3D imaging approaches employ focal stack collection to image different planes of the cell. In this work, we present the design and fabrication of a slanted channel microfluidic chip for 3D fluorescence imaging of cells in flow. The approach employs slanted microfluidic channels fabricated in glass using ultrafast laser inscription. The slanted nature of the microfluidic channels ensures that samples come into and go out of focus, as they pass through the microscope imaging field of view. This novel approach enables the collection of focal stacks in a straight-forward and automated manner, even with off-the-shelf microscopes that are not equipped with any motorized translation/rotation sample stages. The presented approach not only simplifies conventional focal stack collection, but also enhances the capabilities of a regular widefield fluorescence microscope to match the features of a sophisticated confocal microscope. We demonstrate the retrieval of sectioned slices of microspheres and cells, with the use of computational algorithms to enhance the signal-to-noise ratio (SNR) in the collected raw images. The retrieved sectioned images have been used to visualize fluorescent microspheres and bovine sperm cell nucleus in 3D while using a regular widefield fluorescence microscope. We have been able to achieve sectioning of approximately 200 slices per cell, which corresponds to a spatial translation of similar to 15 nm per slice along the optical axis of the microscope.

Item Type: Journal Article
Additional Information: Copy right for this article belongs to the OPTICAL SOC AMER, 2010 MASSACHUSETTS AVE NW, WASHINGTON, DC 20036 USA
Department/Centre: Division of Physical & Mathematical Sciences > Instrumentation Appiled Physics
Depositing User: Id for Latest eprints
Date Deposited: 07 Dec 2016 06:00
Last Modified: 23 Oct 2018 14:50
URI: http://eprints.iisc.ac.in/id/eprint/55557

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