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Biomed Opt Express. 2014 Apr 18;5(5):1588-609. doi: 10.1364/BOE.5.001588. eCollection 2014 May 1.

Optimal lens design and use in laser-scanning microscopy.

Author information

1
Integrative Neurophysiology, Centre for Neurogenomics and Cognitive Research, VU University, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands ; Neuroscience Campus Amsterdam, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands.
2
Integrative Neurophysiology, Centre for Neurogenomics and Cognitive Research, VU University, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands ; Neuroscience Campus Amsterdam, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands ; h.d.mansvelder@vu.nl.

Abstract

In laser-scanning microscopy often an off-the-shelf achromatic doublet is used as a scan lens which can reduce the available diffraction-limited field-of-view (FOV) by a factor of 3 and introduce chromatic aberrations that are scan angle dependent. Here we present several simple lens designs of superior quality that fully make use of high-NA low-magnification objectives, offering diffraction-limited imaging over a large FOV and wavelength range. We constructed a two-photon laser-scanning microscope with optimized custom lenses which had a near diffraction limit point-spread-function (PSF) with less than 3.6% variation over a 400 µm FOV and less than 0.5 µm lateral color between 750 and 1050 nm.

KEYWORDS:

(180.0180) Microscopy; (180.1790) Confocal microscopy; (180.4315) Nonlinear microscopy; (220.3620) Lens system design; (220.3630) Lenses

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