title: Automated spherical aberration correction in scanning confocal microscopy.
abstract: Mismatch between the refractive indexes of immersion media and glass coverslips introduces spherical aberrations in microscopes especially for high numerical aperture objectives. This contribution demonstrates an automated adjustment of the coverslip correction collar in scanning confocal microscopy to compensate for spherical aberrations due to coverslip thickness mismatch. With a motorized coverslip correction collar, the adjustment procedure consists of xz image scans, image processing, correction quality evaluation, the mismatch estimation, and eventually the optimal adjustment of the correction collar. For fast correction with less photodamage, coarse-fine Gaussian fitting algorithms are proposed and evaluated with various specimen for their estimation accuracy. The benefits of the proposed automated correction are demonstrated for various coverslips with biological specimens, showing the optimized resolution of the confocal microscope.
authors.affiliation: Delft Center for Systems and Control, Delft University of Technology, Mekelweg 2, 2628 CD Delft, The Netherlands.
authors.forename: H W
authors.initials: HW
authors.lastname: Yoo
authors.affiliation: Erasmus Optical Imaging Center, Department of Pathology, Josephine Nefkens Institute, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GE, Rotterdam, The Netherlands.
authors.forename: M E
authors.initials: ME
authors.lastname: van Royen
authors.affiliation: Erasmus Optical Imaging Center, Department of Pathology, Josephine Nefkens Institute, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GE, Rotterdam, The Netherlands.
authors.forename: W A
authors.initials: WA
authors.lastname: van Cappellen
authors.affiliation: Erasmus Optical Imaging Center, Department of Pathology, Josephine Nefkens Institute, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GE, Rotterdam, The Netherlands.
authors.forename: A B
authors.initials: AB
authors.lastname: Houtsmuller
authors.affiliation: Delft Center for Systems and Control, Delft University of Technology, Mekelweg 2, 2628 CD Delft, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Verhaegen
authors.affiliation: Automation and Control Institute, Vienna University of Technology, Gusshausstr. 27-29, 1040 Vienna, Austria.
authors.forename: G
authors.initials: G
authors.lastname: Schitter
mesh_terms: D000465:Algorithms; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D031405:Convallaria; D004867:Equipment Design; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods; D051379:Mice; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D008962:Models, Theoretical; D016011:Normal Distribution; D055620:Optical Phenomena; D010363:Pattern Recognition, Automated / Q000379:methods*; D027343:Rhizome / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D014867:Water; D006046:Gold