Live Cell Imaging by Single-Shot Common-Path Wide Field-of-View Reflective Digital Holographic Microscope

Abstract

Quantitative phase imaging by digital holographic microscopy (DHM) is a nondestructive and label-free technique that has been playing an indispensable role in the fields of science, technology, and biomedical imaging. The technique is competent in imaging and analyzing label-free living cells and investigating reflective surfaces. Herein, we introduce a new configuration of a wide field-of-view single-shot common-path off-axis reflective DHM for the quantitative phase imaging of biological cells that leverages several advantages, including being less-vibration sensitive to external perturbations due to its common-path configuration, also being compact in size, simple in optical design, highly stable, and cost-effective. A detailed description of the proposed DHM system, including its optical design, working principle, and capability for phase imaging, is presented. The applications of the proposed system are demonstrated through quantitative phase imaging results obtained from the reflective surface (USAF resolution test target) as well as transparent samples (living plant cells). The proposed system could find its applications in the investigation of several biological specimens and the optical metrology of micro-surfaces.

Plant phenotyping relevance

植物細胞を対象に定量位相画像を取得できる新規デジタルホログラフィック顕微鏡を開発しており、植物試料での性能実証も含むため、植物フェノタイピングに利用可能な画像計測法が中心です。

Code and data availability

The paper's only paper-specific public asset is the MDPI supplementary material containing the time-lapse retrieved wrapped phase imaging video of tobacco plant cells (the paper's plant-cell phenotyping measurements). No analysis code, datasets, or trained models are publicly deposited; the Data Availability Statement仅