Spherical Aberration

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Күн бұрын

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: 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

Пікірлер: 4

@tinglianyuan3098 7 ай бұрын

Could not find a better instructional video on spherical aberration than this! Reach Brilliant when deciphering the aberration at the imaging plane.

@MelHugs Жыл бұрын

This is a really clear and crisp explanation of why image quality can often be non-optimal in one sample, and completely non-standardised from sample to sample. Thank you! The partial solutions described are all good when considering the standard microscope slide system we have to work with. I would be very interested in talking to you about a way in which we can consider improving the sample position, during preparation, on the microscope slide rather than the partial solutions using the microscope.

@MRchalkie666 Жыл бұрын

Nice instructional video! Perhaps we can say that spherical shaped lenses are easier to manufacture and polish, and so they are used, but they are the wrong shape to focus to a point. This is why it's called spherical aberration. Large aspherical lenses are possible to make and are the right shape, but the tiny ones needed on objective lenses are super hard to make and so are not used, and so extra lenses in the objective are used to cancel out the spherical aberration from the spherical top lens.

@Microcourses Жыл бұрын

Thank you for your comment and clear explanation!