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Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

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Published on: April 9, 2014

Interferometric backward third harmonic generation microscopy for axial imaging with accuracy beyond the diffraction

Daaf Sandkuijl¹, Lukas Kontenis¹, Nuno M Coelho²

¹Department of Physics and Institute for Optical Sciences, University of Toronto, Toronto, Ontario, Canada; Department of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, Ontario, Canada.

|April 9, 2014

Summary

This summary is machine-generated.

A novel interferometric microscopy technique precisely measures nanoscale height variations and layer thickness. This advanced imaging method achieves accuracy far beyond the diffraction limit for biological and material surfaces.

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Area of Science:

Nonlinear optics
Microscopy
Nanotechnology

Background:

Accurate measurement of nanoscale variations is crucial in materials science and biology.
Existing microscopy techniques face limitations in resolution and accuracy for certain applications.

Purpose of the Study:

To introduce a new nonlinear microscopy technique for high-accuracy height and thickness measurements.
To demonstrate the technique's capability in visualizing fine surface and layer variations.

Main Methods:

Development of an interferometric technique utilizing backward-reflected third harmonic generation (I-THG) from multiple interfaces.
Application of the I-THG microscope for imaging patterned glass surfaces and fibroblast cell thickness.

Main Results:

Achieved measurement accuracy of up to 5 nm for height variations and layer thickness.
Visualized height variations with accuracy exceeding the diffraction limit by two orders of magnitude.
Demonstrated visualization of thickness variations in fibroblasts.

Conclusions:

The I-THG microscopy technique offers unprecedented accuracy for nanoscale measurements.
This method has broad applications in biological tissue imaging and surface analysis.
The technique surpasses the diffraction limit for precise distance variation measurements.