Enhanced epifluorescence microscopy by uniform and intensity optimized illumination
Perspective published in Cytometry Part A, January 2012
Publisher: Wiley Subscription Services, Inc., A Wiley Company
The manuscript by Sebastien Herbert and Ricardo Henriques discusses the limitations of epifluorescence microscopy due to nonhomogeneous light excitation and the inconsistent illumination problem. They highlight the impact of this issue on image quality and quantification, particularly in techniques like FRET. The authors then introduce a solution for mercury arc lamp-based epifluorescence microscopes, which involves using a dual microlens array to optimize and flatten the illumination pattern. This leads to improved image analysis, better stitching quality, and enhanced assessment and quantification of fluorescence. The authors also mention the challenges of achieving uniform illumination in three-dimensional space and time, and the potential of adaptive optics methods to address this issue. Overall, the manuscript proposes a simple, static, and stable solution for improving the performance of common epifluorescence microscopes.