Evaluating optical aberrations using fluorescent microspheres: methods, analysis, and corrective actions.

Obtaining optimal performance from a microscopy system requires careful evaluation of the entire optics train of the imaging system. This evaluation starts with a fundamental evaluation of the optical components in the microscope. Concise and visual methods are provided for understanding the optical performance of the microscope as a system using subdiffraction fluorescent microspheres to evaluate both monochromatic and polychromatic aberrations. Further practical guides are given to troubleshooting optical problems and final comments are made on optimizing sample preparation.