Characterization of a reflective objective with multiphoton microscopy

Mohammad M. Kabir, Aakash M. Choubal, Mayandi Sivaguru, Kimani C. Toussaint

Research output: Chapter in Book/Report/Conference proceedingConference contribution


Reflective objectives (ROs) can reduce chromatic aberration across a wide wavelength range in multiphoton microscopy (MPM). However, a systematic characterization of the performance of ROs has not been carried out. In this paper, we analyze the performance of a 0.5 numerical-aperture (NA) RO and compare it with a 0.55 NA standard glass objective (SO), using two-photon fluorescence (TPF) and second-harmonic generation (SHG). For experiments extending ∼1 octave in visible and NIR wavelengths, the SO introduces defocusing errors of ∼25% for TPF images of sub-diffraction fluorescent beads and ∼10% for SHG images of collagen fibers. For both imaging systems, the RO provides a corresponding error of ∼4%. This work highlights the potential usefulness of ROs for multimodal MPM applications.

Original languageEnglish (US)
Title of host publicationMultiphoton Microscopy in the Biomedical Sciences XVIII
EditorsAmmasi Periasamy, Xiaoliang S. Xie, Xiaoliang S. Xie, Karsten Konig, Peter T. C. So
ISBN (Electronic)9781510614819
StatePublished - 2018
EventMultiphoton Microscopy in the Biomedical Sciences XVIII 2018 - San Francisco, United States
Duration: Jan 28 2018Jan 30 2018

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
ISSN (Print)1605-7422


OtherMultiphoton Microscopy in the Biomedical Sciences XVIII 2018
Country/TerritoryUnited States
CitySan Francisco


  • multiphoton microscopy
  • reflective objective
  • second-harmonic generation
  • two-photon fluorescence

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging
  • Biomaterials


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