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.
Highlights
Light-sheet microscopy enables considerable speed and phototoxicity gains, while quantitativephase imaging confers label-free recognition of cells and organelles, and quantifies their numberdensity that, thermodynamically, is more representative of metabolism than size
While quantitative-phase imaging (QPI) is compatible with most sample mounting techniques and microfluidics, light-sheet imaging (LSI) and LLSI generally require practices that are atypical to common cell culture techniques or are prone to contamination
This is pertinent to QPI since it relies on the interference between the transmitted field through the cell and a reference field[14]
Summary
Light-sheet microscopy enables considerable speed and phototoxicity gains, while quantitativephase imaging confers label-free recognition of cells and organelles, and quantifies their numberdensity that, thermodynamically, is more representative of metabolism than size. Quantitative-phase imaging (QPI) retains low phototoxicity and confers label-free recognition and number-density quantification of single cells and their organelles[10,11,12,13,14] These two imaging modalities offer complementary advantages. Single-objective configurations can potentially alleviate any spatial frequency or energy losses and enable compatibility with common sample-mounting techniques To this end, several single-objective configurations have been recently reported, with some enabling extremely large fields-of-view (FOVs)[19,20,21,22]; approaches that enable submicron (planar/axial) resolution, such as epi-illumination SPIM or highly inclined and laminated optical sheet (HILO)[6,7,8], typically achieve only a fraction of the field-of-view (FOV) endowed by QPI This FOV discrepancy at submicron (or subcellular) resolution levels can yield significant differences between the throughput rates of QPI and LSI (or LLSI), which can be detrimental to statistically significant investigations at the single-cell level
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