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Application of a reflective microscope objective for multiphoton microscopy.

Mohammad M Kabir1, Aakash M Choubal2, Kimani C Toussaint2

  • 1Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

Journal of Microscopy
|April 21, 2018
PubMed
Summary
This summary is machine-generated.

Reflective objectives (ROs) significantly reduce optical errors in multiphoton microscopy compared to standard objectives. This makes ROs ideal for advanced imaging applications requiring high fidelity across various wavelengths.

Keywords:
Multiphoton microscopyreflective objectivesecond-harmonic generationtwo-photon fluorescence

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Area of Science:

  • Optical microscopy
  • Biomedical imaging
  • Materials science

Background:

  • Chromatic aberration limits standard objectives across broad wavelengths.
  • Systematic performance characterization of reflective objectives (ROs) is lacking.
  • ROs offer potential for advanced optical microscopy.

Purpose of the Study:

  • Compare the performance of ROs and standard objectives (SOs).
  • Evaluate imaging fidelity using two-photon fluorescence (TPF) and second-harmonic generation (SHG).
  • Assess performance across visible and near-infrared (NIR) wavelengths.

Main Methods:

  • Utilized a 0.5 NA RO and a 0.55 NA SO.
  • Conducted experiments using TPF imaging of subdiffraction fluorescent beads.
  • Performed SHG imaging of collagen fibers.
  • Covered a wavelength range of approximately one octave in the visible and NIR spectrum.

Main Results:

  • SOs exhibited significant defocusing errors (25-40% for TPF, 10-12% for SHG).
  • ROs demonstrated minimal defocusing errors (approximately 4% for both TPF and SHG).
  • ROs maintained high imaging fidelity across the tested wavelength range.

Conclusions:

  • ROs effectively mitigate chromatic aberration and defocusing errors.
  • ROs show significant advantages over SOs for multimodal multiphoton microscopy.
  • ROs are highly suitable for applications demanding broad wavelength compatibility and precision.