Light Microscopy Research Group (LMRG): International Test Results for Objective Lens Quality Resolution Spectral Accuracy and Spectral Separation for Confocal Laser Scanning Microscopes (CLSM)

摘要

As part of an ongoing effort to increase image reproducibility and fidelity in addition to improving cross-instrument consistency we have developed four separate instrument quality tests to augment the ones previously reported. 1) Objective lens quality 2) Resolution 3) Spectral accuracy of the wavelength information from spectral detectors 4) Spectral Separation tested the accuracy and quality of un-mixing algorithms. To ascertain the usefulness of these tests as well as to determine the current ““state”” of microscopes in use, test specimens and detailed protocols were made available worldwide free-of-charge. 55 laboratories located in 18 countries provided data. Objective lens quality: good with most issues arising from user errors or stage/focus drift, approximately 10% of lenses had aberrations. Resolution: within an average of approximately 25% of theoretical values. Spectral accuracy: excellent, even for low resolution systems. Spectral un-mixing: good, poor data collection was the main cause of low quality data.