ABRF Light Microscopy Research Group Study 3: the Development and Implementation of New Tools for Microscope Quality Assurance Testing.

The goal of this study was to create a 3D biologically-relevant standard test sample and imaging protocol which would allow a microscopist to evaluate the performance of their imaging system as a function of imaging depth in the following areas: (1) system resolution and point spread function quality in 3D, (2) reliability of intensity quantification, and (3) image signal-to-noise. We developed two test samples that each consist of a mixture of fluorescence microspheres embedded in a 100µm-thick layer of CyGel, a thermo-reversible 3D cell growth media with refractive index of 1.37. We will present a summary of our methodology and quantitative analysis of approximately 140 laser scanning confocal datasets collected from over 50 participants across 15 countries and including instruments from four major microscope manufacturers. The results show some interesting and unexpected trends and provide tools and benchmarks that investigators can use to evaluate systems at their own institutions.