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Spherical Aberration

Spherical aberration distorts images generated with a light microscope. In this Microcourse, you'll learn what spherical aberration is, what is does to your images, where it comes from and how to deal with it. Thanks to my fabulous group for feedback on this video prior to release: Talley Lambert, Anna Jost, Rylie Walsh & Federico Gasparoli. References & Resources: Arimoto, R. and Murray, J.M. (2004) A common aberration with water-immersion objective lenses. J. Microsc. 216, 49–51 Goodwin, P.C. (2013) Evaluating optical aberrations using fluorescent microspheres: methods, analysis, and corrective actions. Methods Cell Biol. 114, 369–385 Hell, S. et al. (2011) Aberrations in confocal fluorescence microscopy induced by mismatches in refractive index. J. Microsc. 169, 391–405 Hiraoka, Y. et al. (1990) Determination of three-dimensional imaging properties of a light microscope system. Partial confocal behavior in epifluorescence microscopy. Biophys. J. 57, 325–333 Joglekar, A.P. et al. (2008) Counting kinetochore protein numbers in budding yeast using genetically encoded fluorescent proteins. Methods Cell Biol. 85, 127–151 Ross, S.T. et al. (2014) Practical considerations of objective lenses for application in cell biology. Methods Cell Biol. 123, 19–34 Waters, J.C. (2007) Live‐Cell Fluorescence Imaging. Methods Cell Biol. 81, 115–140 Fluorescence mounting media suggestions: https://nic.med.harvard.edu/resources... Chapters: 00:00 Introduction to spherical aberration 02:11 Identifying spherical aberration 02:48 Effects on image quality 04:36 Causes of spherical aberration: Optics 06:07 Causes of spherical aberration: Specimens 08:34 Why specimens cause spherical aberration 12:22 Spherical aberration with depth 12:51 Effect on intensity 14:00 Solutions for spherical aberration 14:38 Easier solutions 19:06 More complicated solutions 20:34 All solutions are limited