Dynamic quantitative phase imaging using calcite crystal-based temporally stable interferometer

Abstract

This paper describes the development of a common path off-axis quantitative phase microscope (QPM), enabling high-speed quantitative measurements of sub-cellular dynamics in a live cell. The proposed 4f optical system consists of a polarizer, lenses, calcite crystal, pinhole and analyser, attached to the output port of an optical microscope converting it into a temporally stable QPM. The temporal phase stability of this optical configuration is measured to be equal to ∼20 mrad without a vibration isolation table. The utility of the microscope is demonstrated by conducting experiments on a polystyrene sphere (dia. 12 μm) and human red blood cells (RBCs). Furthermore, the developed system is implemented to study temporal membrane fluctuation in human RBCs over ∼30s and mouse embryonic fibroblast cell dynamics over 2 hours. Therefore, the system offers an energy-efficient and simple solution for generating object and reference beams for common-path QPM, making it an attractive alternative to existing approaches.

KEYWORDS:

• Quantitative phase microscope
• temporal stable interferometer
• cell dynamics
• bio imaging

Author contributions

A.A. has conceptualized the idea and designed the experiments. A.S. performed the experiments and analysed the data. V.D. and A.H. assisted during the experiments and prepared bio-samples. A.S. and A.A. mainly wrote the manuscript. B.S.A., and D.S.M. conceived the project. A.A., B.S.A., D.S.M., and SKD supervised this work. All authors reviewed and edited the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The authors declare the availability of the data used in the research to obtain the results reported in the manuscript upon reasonable request.

Additional information

Funding

B.S.A. and DSM acknowledge the funding from the INTPART [project # 309802]. BSA and AA acknowledge the funding from Research Council of Norway, project NANO 2021-288565. AA acknowledge the funding from the Ministry of Human Resources and Development (MHRD), Govt. of India.