Characterization of a reflective objective with multiphoton microscopy

Mohammad M. Kabir; Aakash M. Choubal; Mayandi Sivaguru; Kimani C. Toussaint Jr.

doi:10.1117/12.2290305

23 February 2018 Characterization of a reflective objective with multiphoton microscopy

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

Proceedings Volume 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII; 104982Z (2018) https://doi.org/10.1117/12.2290305
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

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.

Citation Download Citation

Mohammad M. Kabir, Aakash M. Choubal, Mayandi Sivaguru, and Kimani C. Toussaint Jr. "Characterization of a reflective objective with multiphoton microscopy", Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 104982Z (23 February 2018); https://doi.org/10.1117/12.2290305

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