ConfocalCheck - A Software Tool for the Automated Monitoring of Confocal Microscope Performance

• Published in: PloS one Vol. 8; no. 11; p. e79879
• Main Authors: Hng, Keng Imm, Dormann, Dirk
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 05.11.2013; Public Library of Science (PLoS)
• Subjects: Automation; Computer programs; Confocal microscopy; Convallaria; Data processing; Diagnostic systems; Experiments; Image acquisition; Image processing; Image quality; Lasers; Light; Measuring instruments; Microscopes; Microscopy; Microscopy, Confocal - methods; Monitoring instruments; Performance assessment; Point spread functions; Quality assurance; Quality control; Software; Software development tools; Software industry; Web sites; Websites; United Kingdom > UK; Fiji
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0079879

Laser scanning confocal microscopy has become an invaluable tool in biomedical research but regular quality testing is vital to maintain the system's performance for diagnostic and research purposes. Although many methods have been devised over the years to characterise specific aspects of a confocal microscope like measuring the optical point spread function or the field illumination, only very few analysis tools are available. Our aim was to develop a comprehensive quality assurance framework ranging from image acquisition to automated analysis and documentation. We created standardised test data to assess the performance of the lasers, the objective lenses and other key components required for optimum confocal operation. The ConfocalCheck software presented here analyses the data fully automatically. It creates numerous visual outputs indicating potential issues requiring further investigation. By storing results in a web browser compatible file format the software greatly simplifies record keeping allowing the operator to quickly compare old and new data and to spot developing trends. We demonstrate that the systematic monitoring of confocal performance is essential in a core facility environment and how the quantitative measurements obtained can be used for the detailed characterisation of system components as well as for comparisons across multiple instruments.