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Related Experiment Video

Updated: May 2, 2026

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers
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Phase-gradient contrast in thick tissue with a scanning microscope.

J Mertz1, A Gasecka2, A Daradich2

  • 1Boston University, Department of Biomedical Engineering, 44 Cummington St., Boston, MA 02215 USA.

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|February 28, 2014
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Summary
This summary is machine-generated.

A new scanning oblique back-illumination microscopy (sOBM) technique provides sub-surface images of unlabeled tissue. This cost-effective add-on offers complementary morphological information for various microscopy methods, operating at video rate.

Keywords:
(110.0180) Microscopy(180.5810) Scanning microscopy(290.7050) Turbid media

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Area of Science:

  • Biomedical Optics
  • Microscopy Techniques
  • Biophotonics

Background:

  • The principle of reciprocity is fundamental for light propagation in scattering and absorptive media.
  • Reciprocity has enabled equivalences between different microscopy modalities.
  • Existing techniques may lack sub-surface imaging capabilities for thick, unlabeled tissues.

Purpose of the Study:

  • To introduce a scanning version of oblique back-illumination microscopy (sOBM).
  • To demonstrate sOBM's capability for sub-surface phase-gradient and amplitude imaging.
  • To showcase sOBM as a versatile, cost-effective add-on for scanning microscopes.

Main Methods:

  • Utilized the principle of optical reciprocity.
  • Developed a scanning oblique back-illumination microscopy (sOBM) technique.
  • Integrated sOBM with two-photon excited fluorescence (TPEF) and coherent anti-Stokes Raman scattering (CARS) microscopy.
  • Applied the technique to brain and spinal cord tissue imaging in an epi-detection geometry.

Main Results:

  • sOBM successfully generated sub-surface phase-gradient and amplitude images of unlabeled tissue.
  • The technique provided complementary morphological information to TPEF and CARS contrast.
  • Simultaneous and automatically co-registered imaging was achieved.
  • sOBM demonstrated operation at video rate.

Conclusions:

  • sOBM is a valuable technique for imaging arbitrarily thick, unlabeled tissues.
  • It offers complementary morphological insights, enhancing existing microscopy methods like TPEF and CARS.
  • The simplicity and cost-effectiveness make sOBM a practical add-on for various scanning microscopes.