Integrative quantitative-phase and airy light-sheet imaging

Scientific Reports
N R SubediA. E. Vasdekis

Abstract

Light-sheet microscopy enables considerable speed and phototoxicity gains, while quantitative-phase imaging confers label-free recognition of cells and organelles, and quantifies their number-density that, thermodynamically, is more representative of metabolism than size. Here, we report the fusion of these two imaging modalities onto a standard inverted microscope that retains compatibility with microfluidics and open-source software for image acquisition and processing. An accelerating Airy-beam light-sheet critically enabled imaging areas that were greater by more than one order of magnitude than a Gaussian beam illumination and matched exactly those of quantitative-phase imaging. Using this integrative imaging system, we performed a demonstrative multivariate investigation of live-cells in microfluidics that unmasked that cellular noise can affect the compartmental localization of metabolic reactions. We detail the design, assembly, and performance of the integrative imaging system, and discuss potential applications in biotechnology and evolutionary biology.

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Methods Mentioned

BETA
light interference microscopy

Software Mentioned

Micro
MicroManager
Matlab
CellVista
OriginPro Pro
ImageJ
OriginLab
Manager
QPI
DeconvolutionLab2 ImageJ

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