Cells grown in a monolayer tend to flatten in the lower part of the plate adhering to and spreading in the horizontal plane without expanding in the vertical dimension. The result is that cells grown in 2D have a forced apex-basal polarity. Microfluidic Live-Imaging with CellViewer technology is an ideal solution to observe the maintenance of a cell in excellent health, trying to bridge the gap between the 2D and 3D model. In this work we propose to test the system on a single isolated Jurkat cell in the microfluidic cartridge and record the timelapse for 4 hours. After adaptive autofocus, when sliding inside the cartridge chamber, the single cell is tracked under the action of the optics and the 3D rotation was experimentally successfully achieved. Then a single cell viability assessment was used using MitoGreen-dye a fluorescence marker selectively permeable to live cells. ImageJ software was used to: calculate the diameter of a single cell, create fluorescence intensity graphs along a straight line passing through the cell, visualize spatial fluorescence intensity distribution in 3D.

MICROFLUIDIC LIVE-IMAGING WITH CELLVIEWER TECHNOLOGY TO PERFORM BIOTECHNOLOGICAL TASKS

Capuzzo Arnaud Martino
Investigation
;
2020-01-01

Abstract

Cells grown in a monolayer tend to flatten in the lower part of the plate adhering to and spreading in the horizontal plane without expanding in the vertical dimension. The result is that cells grown in 2D have a forced apex-basal polarity. Microfluidic Live-Imaging with CellViewer technology is an ideal solution to observe the maintenance of a cell in excellent health, trying to bridge the gap between the 2D and 3D model. In this work we propose to test the system on a single isolated Jurkat cell in the microfluidic cartridge and record the timelapse for 4 hours. After adaptive autofocus, when sliding inside the cartridge chamber, the single cell is tracked under the action of the optics and the 3D rotation was experimentally successfully achieved. Then a single cell viability assessment was used using MitoGreen-dye a fluorescence marker selectively permeable to live cells. ImageJ software was used to: calculate the diameter of a single cell, create fluorescence intensity graphs along a straight line passing through the cell, visualize spatial fluorescence intensity distribution in 3D.
2020
Microfluidics; Biotechnology; Live-Imaging; Biomedical; Biomedical Enineering
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/1088432
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