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

Published online by Cambridge University Press:  06 February 2019

Anna H. Klemm Open the ORCID record for Anna H. Klemm [Opens in a new window] ,
Andreas W. Thomae Open the ORCID record for Andreas W. Thomae [Opens in a new window] ,
Katarina Wachal  and
Steffen Dietzel Open the ORCID record for Steffen Dietzel [Opens in a new window]
Anna H. Klemm*
Affiliation:
Core Facility Bioimaging at the Biomedical Center and Walter-Brendel-Zentrum für Experimentelle Medizin, Ludwig-Maximilians-Univeristät München, Großhaderner Straße 9, 82152 Planegg-Martinsried, Germany
Andreas W. Thomae
Affiliation:
Core Facility Bioimaging at the Biomedical Center and Walter-Brendel-Zentrum für Experimentelle Medizin, Ludwig-Maximilians-Univeristät München, Großhaderner Straße 9, 82152 Planegg-Martinsried, Germany
Katarina Wachal
Affiliation:
Core Facility Bioimaging at the Biomedical Center and Walter-Brendel-Zentrum für Experimentelle Medizin, Ludwig-Maximilians-Univeristät München, Großhaderner Straße 9, 82152 Planegg-Martinsried, Germany
Steffen Dietzel*
Affiliation:
Core Facility Bioimaging at the Biomedical Center and Walter-Brendel-Zentrum für Experimentelle Medizin, Ludwig-Maximilians-Univeristät München, Großhaderner Straße 9, 82152 Planegg-Martinsried, Germany
*
*Author for correspondence: Steffen Dietzel, E-mail: dietzel@lmu.de; Anna H. Klemm, E-mail: anna.klemm@it.uu.se
*Author for correspondence: Steffen Dietzel, E-mail: dietzel@lmu.de; Anna H. Klemm, E-mail: anna.klemm@it.uu.se
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Abstract

Routine system checks are essential for supervising the performance of an advanced light microscope. Recording and evaluating the point spread function (PSF) of a given system provides information about the resolution and imaging. We compared the performance of fluorescent and gold beads for PSF recordings. We then combined the open-source evaluation software PSFj with a newly developed KNIME pipeline named PSFtracker to create a standardized workflow to track a system's performance over several measurements and thus over long time periods. PSFtracker produces example images of recorded PSFs, plots full-width-half-maximum (FWHM) measurements over time and creates an html file which embeds the images and plots, together with a table of results. Changes of the PSF over time are thus easily spotted, either in FWHM plots or in the time series of bead images which allows recognition of aberrations in the shape of the PSF. The html file, viewed in a local browser or uploaded on the web, therefore provides intuitive visualization of the state of the PSF over time. In addition, uploading of the html file on the web allows other microscopists to compare such data with their own.

Keywords

full-width-half-maximumgold beadsKNIMElight microscopymicroscope quality controlpoint spread functionPSFj
Type
Software and Instrumentation
Information
Microscopy and Microanalysis , Volume 25 , Issue 3 , June 2019 , pp. 699 - 704
Copyright
Copyright © Microscopy Society of America 2019 

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Footnotes

Current address: Anna Klemm, Department of Information Technology, Division of Visual Information and Interaction and Science for Life Laboratory, Uppsala University, Sweden. anna.klemm@it.uu.se, Postal address: Box 337, 751 05 UPPSALA. Phone: +46/18-471 2868

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