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Transport-of-intensity phase imaging using commercially available confocal microscope.

Naru Yoneda^1,2, Joe Sakamoto^3,4, Takumi Tomoi^5,6,7

¹Kobe University, Graduate School of System Informatics, Department of System Science, Kobe, Japan.

Journal of Biomedical Optics

|November 8, 2024

Summary

This summary is machine-generated.

Researchers transformed standard confocal microscopes into phase measurement microscopes using the transport of intensity equation (TIE). This innovation enables quantitative phase imaging (QPI) in existing instruments, offering new insights into cell biology.

Keywords:

confocal microscopy fluorescence imaging fluorescence microscopy microscopy quantitative phase imaging transport of intensity equation

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Area of Science:

Biophysics
Optical Microscopy
Cell Biology

Background:

Confocal microscopy is vital for high-resolution biological imaging, particularly for fluorescence in living cells.
Phase information offers valuable insights, including physical parameter measurement and wavefront correction, but is absent in commercial confocal systems.
Existing confocal microscopes lack integrated phase imaging capabilities.

Purpose of the Study:

To adapt commercially available confocal microscopes for quantitative phase imaging (QPI).
To introduce phase imaging functionality without requiring hardware modifications to existing microscopes.
To enable biologists to measure phase information using their current confocal microscopy setups.

Main Methods:

Phase imaging was implemented using the transport of intensity equation (TIE).
The method leverages the bright-field imaging module of standard confocal microscopes.
No physical modifications to the off-the-shelf confocal microscope were necessary.

Main Results:

The feasibility of TIE-based QPI was successfully demonstrated on a microlens array.
Phase imaging was validated using living plant cells (moss) and mammalian cultured cells.
Multimodal imaging combining fluorescence and phase information was achieved.

Conclusions:

Quantitative phase imaging (QPI) is achievable with commercially available confocal microscopes using the TIE technique.
This method provides a non-invasive way to measure physical properties like dry mass, viscosity, and temperature in cells.
Future applications include aberration correction and scattering cancellation through phase fluctuation compensation.