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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Related Experiment Video

Updated: Jul 1, 2026

Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model
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Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model

Published on: September 30, 2021

Real-time robust autofocus method enabling sustained intravital scanning light field imaging.

Yuedi Wang1, Jingyao Wu2,3, Jiamin Wu3,4,5

  • 1School of Information and Communication Engineering, Communication University of China, Beijing, China.

Nature Communications
|June 29, 2026
PubMed
Summary
This summary is machine-generated.

We developed a real-time autofocus method for scanning light-field microscopy (AFsLF) to maintain focus during long-term, high-speed 3D imaging of living samples, overcoming challenges from tissue movement.

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Last Updated: Jul 1, 2026

Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model
07:50

Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model

Published on: September 30, 2021

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers
10:07

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Published on: April 9, 2014

Area of Science:

  • Biomedical imaging
  • Microscopy techniques
  • Computational imaging

Background:

  • Intravital 3D imaging faces challenges with sample stability due to vibration and deformation.
  • Maintaining focus over extended periods is difficult, limiting long-term live imaging.
  • Existing methods struggle with sustained, high-resolution imaging in dynamic biological systems.

Purpose of the Study:

  • To introduce a real-time autofocus method for sustained, high-speed 3D intravital imaging.
  • To enable stable, long-term imaging of diverse biological samples without hardware modifications.
  • To overcome limitations in focus tracking for dynamic tissues.

Main Methods:

  • Developed a real-time robust autofocus method based on scanning light-field microscopy (AFsLF).
  • Utilized intrinsic disparity of light-field angular measurements for focal plane estimation.
  • AFsLF operates without requiring hardware modifications to the microscope.

Main Results:

  • AFsLF achieves sustained high-speed 3D imaging of diverse samples over several days.
  • Focal plane estimation error is less than 2 µm over a 500 µm depth range.
  • The method completes autofocus within 0.1 s, significantly faster than previous approaches.
  • Validated AFsLF under challenging conditions, including low excitation power and multichannel illumination.

Conclusions:

  • AFsLF provides a robust solution for stable, long-term, high-speed 3D intravital imaging.
  • Enables unprecedented subcellular imaging of neural activities and immune responses in vivo.
  • Represents a significant advancement for studying dynamic biological processes in live organisms.