Application of a reflective microscope objective for multiphoton microscopy

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

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)129-135
Number of pages7
JournalJournal of Microscopy
Volume271
Issue number2
DOIs
StatePublished - Aug 2018

Keywords

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

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

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