Integrated Confocal Performance Assessment

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Poster Session 2
Alfonso J. Schmidt (1), Kylie M. Price (1), Graham D. Wright (2)
1. 1) Malaghan Institute of Medical Research, Hugh Green Cytometry Centre, Wellington New Zealand.
2. 2) A*STAR Microscopy Platform (AMP), Research Support Centre, Agency for Science, Technology & Research (A*STAR), Singapore.

performance assessment, laser scanning confocal microscope, fluorescence, microscopy calibration, beads, co-registration, field Illumination, stability 

Abstract text

The creation of standard operating procedures for the evaluation of instrument performance, and their importance, is an area currently gaining traction within the microscopy community globally.  It is being led by worldwide initiatives like QUAREP-LiMi [1] with the intention of providing guidelines for quality assessment and experimental reproducibility. However, current literature around this topic shows quality assessment approaches that evaluate individual components and the results are reported separately which reduces the scope and overall interpretation of the results of the assessment.


Inspired by published assessments and based on experience in the field, six parameters have been selected to create a holistic, evaluation tool to assess confocal microscope performance; (1) Laser power and stability, (2) uniformity of field of view, (3) lateral and axial co-registration, (4) motorized stage precision and z-drift, (5) detector sensitivity and linearity, and (6) point spread function (PSF). Furthermore, we propose a parametrization of these results to provide clear categories (e.g. Optimal, Acceptable and Fail) that will facilitate decision making regarding instrument performance. We propose a spider chart that provides information for each particular evaluated parameter while simultaneously providing the complementary equipment overview perspective.


It is expected that the assessment parametrization and spider chart outputs will provide a useful tool to the field. In a core facility environment, it can be used to provide information to end-users regarding equipment performance expectations, to identify the best match between equipment and experiment goal, it can provide useful performance tracking information to support instrument service management and pre-emptive maintenance. Further is can be used as a tool to aid decision making in the technology purchase and asset lifecycle control, by comparing equipment based on performance and unbiased conditions over time, especially powerful when combined with insights on utilisation dynamics.


This Integrated Confocal Performance Assessment project is being undertaken as the Technical Essay of the Royal Microscopy Society Diploma Program by Alfonso Schmidt.


[1] QUAREP-LiMi: A community-driven initiative to establish guidelines for quality assessment and reproducibility for instruments and images in light microscopy, arXiv:2101.09153 [q-bio.OT]