Objective-Induced Point Spread Function Aberrations and Their Impact on Super-Resolution Microscopy

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• 作者：Karole L. Blythe ; Eric J. Titus ; Katherine A. Willets
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：June 16, 2015
• 年：2015
• 卷：87
• 期：12
• 页码：6419-6424
• 全文大小：358K
• ISSN：1520-6882

文摘

This study demonstrates how different microscope objectives can lead to asymmetric imaging aberrations in the point spread function of dipolar emitters, which can adversely affect the quality of fit in super-resolution imaging. Luminescence from gold nanorods was imaged with four different objectives to measure the diffraction-limited emission and characterize deviations from the expected dipolar emission patterns. Each luminescence image was fit to a three-dipole emission model to generate fit residuals that visually relay aberrations in the point spread function caused by the different microscope objectives. Output parameters from the fit model were compared to experimentally measured values, and we find that while some objectives provide high quality fits across all nanorods studied, others show significant aberrations and are inappropriate for super-resolution imaging. This work presents a simple and robust strategy for quickly assessing the quality of point spread functions produced by different microscope objectives.