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Tracking Microscope Performance: A Workflow to Compare Point Spread Function Evaluations Over Time.

Anna H Klemm1, Andreas W Thomae1, Katarina Wachal1

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Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
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Summary
This summary is machine-generated.

Routine system checks for advanced light microscopes are crucial. A new workflow using PSFtracker and PSFj software standardizes point spread function (PSF) evaluation, enabling long-term performance tracking and aberration detection.

Keywords:
KNIMEPSFjfull-width-half-maximumgold beadslight microscopymicroscope quality controlpoint spread function

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Area of Science:

  • Microscopy
  • Optical Engineering
  • Data Science

Background:

  • Advanced light microscopes require regular performance monitoring.
  • The point spread function (PSF) is a key metric for assessing microscope resolution and imaging quality.

Purpose of the Study:

  • To develop a standardized workflow for long-term tracking of advanced light microscope performance.
  • To compare fluorescent and gold beads for point spread function (PSF) recordings.

Main Methods:

  • Combined open-source PSFj software with a new KNIME pipeline, PSFtracker.
  • Utilized fluorescent and gold beads for PSF recordings.
  • Developed a workflow to track system performance over multiple measurements.

Main Results:

  • PSFtracker generates visualizations, including example PSF images and full-width-half-maximum (FWHM) plots over time.
  • An HTML file summarizes results, facilitating easy identification of PSF aberrations and changes.
  • The workflow enables intuitive visualization and comparison of PSF data over time.

Conclusions:

  • The developed workflow provides a standardized method for monitoring microscope performance.
  • PSFtracker facilitates the detection of aberrations and changes in PSF over extended periods.
  • The generated HTML reports enhance data sharing and comparison among microscopists.