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High-speed multifocus phase imaging in thick tissue.

Sheng Xiao1, Shuqi Zheng1, Jerome Mertz1

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

Biomedical Optics Express
|October 25, 2021
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Summary
This summary is machine-generated.

This study introduces a novel epi-mode, multifocus phase imaging technique for unstained biological samples. The method enables high-speed 3D imaging of thick tissues, capturing dynamic processes like blood flow.

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Area of Science:

  • Biomedical Optics
  • Microscopy
  • Biophotonics

Background:

  • Phase microscopy is crucial for imaging unstained biological samples.
  • Existing techniques are limited to transmission geometries, unsuitable for thick samples.
  • Volumetric phase imaging is often too slow for live biological processes.

Purpose of the Study:

  • To develop a high-speed, 3D phase imaging technique for thick, scattering biological tissues.
  • To overcome the limitations of transmission-based phase microscopy.
  • To enable the study of dynamic biological processes in vivo.

Main Methods:

  • Combined oblique back-illumination microscopy with a z-splitter prism.
  • Implemented an epi-mode, multifocus phase imaging approach.
  • Utilized a single camera for data acquisition.

Main Results:

  • Achieved 3D qualitative phase imaging in thick, scattering tissues.
  • Demonstrated high-speed imaging at up to 47 Hz.
  • Successfully imaged blood flow in chick embryos over a 546 × 546 × 137 µm³ volume.

Conclusions:

  • The developed technique offers a versatile solution for high-speed 3D phase imaging.
  • This advancement is suitable for studying dynamic biological processes in complex tissues.
  • The method expands the applicability of phase microscopy to thick and scattering samples.