Issue 9, 2023

Holographic flow scanning cytometry overcomes depth of focus limits and smartly adapts to microfluidic speed

Abstract

Space-time digital holography (STDH) maps holograms in a hybrid space-time domain to achieve extended field of view, resolution enhanced, quantitative phase-contrast microscopy and velocimetry of flowing objects in a label-free modality. In STDH, area sensors can be replaced by compact and faster linear sensor arrays to augment the imaging throughput and to compress data from a microfluidic video sequence into one single hybrid hologram. However, in order to ensure proper imaging, the velocity of the objects in microfluidic channels has to be well-matched to the acquisition frame rate, which is the major constraint of the method. Also, imaging all the flowing samples in focus at the same time, while avoiding hydrodynamic focusing devices, is a highly desirable goal. Here we demonstrate a novel processing pipeline that addresses non-ideal flow conditions and is capable of returning the correct and extended focus phase contrast mapping of an entire microfluidic experiment in a single image. We apply this novel processing strategy to recover phase imaging of flowing HeLa cells in a lab-on-a-chip platform even when severely undersampled due to too fast flow while ensuring that all cells are in focus.

Graphical abstract: Holographic flow scanning cytometry overcomes depth of focus limits and smartly adapts to microfluidic speed

Supplementary files

Article information

Article type
Paper
Submitted
20 Jan 2023
Accepted
03 Apr 2023
First published
11 Apr 2023
This article is Open Access
Creative Commons BY-NC license

Lab Chip, 2023,23, 2316-2326

Holographic flow scanning cytometry overcomes depth of focus limits and smartly adapts to microfluidic speed

Z. Wang, V. Bianco, P. L. Maffettone and P. Ferraro, Lab Chip, 2023, 23, 2316 DOI: 10.1039/D3LC00063J

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