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Application of a reflective microscope objective for multiphoton microscopy
Author(s) -
KABIR MOHAMMAD M.,
CHOUBAL AAKASH M.,
TOUSSAINT KIMANI C
Publication year - 2018
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12702
Subject(s) - optics , chromatic aberration , microscopy , multiphoton fluorescence microscope , microscope , materials science , wavelength , fluorescence , second harmonic generation , fluorescence microscope , numerical aperture , octave (electronics) , chromatic scale , optical microscope , optoelectronics , physics , scanning electron microscope , laser
Summary Reflective objectives (ROs) mitigate chromatic aberration across a broad wavelength range. Yet, a systematic performance characterisation of ROs has not been done. In this paper, we compare the performance of a 0.5 numerical‐aperture (NA) reflective objective (RO) with a 0.55 NA standard glass objective (SO), using two‐photon fluorescence (TPF) and second‐harmonic generation (SHG). For experiments spanning ∼1 octave in the visible and NIR wavelengths, the SO leads to defocusing errors of 25–40% for TPF images of subdiffraction fluorescent beads and 10–12% for SHG images of collagen fibres. The corresponding error for the RO is ∼4% for both imaging modalities. This work emphasises the potential utility of ROs for multimodal multiphoton microscopy applications.