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Active focus locking in an optically sectioning microscope utilizing a deformable membrane mirror.

S P Poland¹, A J Wright, J M Girkin

¹Institute of Photonics, SUPA, University of Strathclyde, Glasgow, Scotland, UK. simon.poland@strath.ac.uk

|March 4, 2008

Summary

Movement artifacts in high-resolution microscopy hinder in vivo imaging. A novel actively locked focus-tracking system using a deformable membrane mirror effectively compensates for axial movements, improving image quality.

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Area of Science:

Biomedical imaging
Optical microscopy
Instrumentation

Background:

In vivo imaging faces significant challenges from movement artifacts, primarily axial, affecting high-resolution techniques like confocal and multiphoton microscopy.
These movements, caused by physiological processes or sample responses, displace features out of the focal plane, degrading image quality.

Purpose of the Study:

To develop and evaluate an actively locked focus-tracking system to mitigate movement artifacts in high-resolution in vivo imaging.
To demonstrate the system's effectiveness and operational limits in simulated biological applications.

Main Methods:

Development of an actively locked focus-tracking system utilizing a deformable membrane mirror.
Simulation of sample oscillation and movement to test the focus-locking system.
Evaluation of tracking range and precision in simulated biological scenarios.

Main Results:

The focus-tracking system successfully compensated for axial movements.
A focus tracking range of 400 micrometers was achieved, limited by the objective's piezoelectric mount.
A root-mean-square precision of 0.31 microm +/- 0.05 microm on focal depth was demonstrated.

Conclusions:

The deformable membrane mirror-based focus-tracking system offers an effective solution for movement artifacts in high-resolution microscopy.
This active system provides an advantage over conventional methods by avoiding direct sample contact.
The technology shows promise for various in vivo imaging applications requiring stable focal depth.