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Imaging Biological Samples with Optical Microscopy01:18

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
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Related Experiment Video

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Enhancing Total Optical Throughput of Microscopy with Deep Learning for Intravital Observation.

Runze Chen1, Shiyi Peng1, Liang Zhu2

  • 1College of Optical Science and Engineering, State Key Laboratory of Modern Optical Instrumentations, International Research Center for Advanced Photonics, Centre for Optical and Electromagnetic Research, Zhejiang University, 310058, Hangzhou, China.

Small Methods
|May 15, 2023
PubMed
Summary

Researchers developed an advanced near-infrared-II (NIR-II) microscope for high-speed, high-resolution in vivo imaging. This breakthrough enables detailed visualization of tiny brain vessels, crucial for understanding diseases.

Keywords:
NIR-II microscopeenhanced spatial resolutionhigh optical throughputhigh temporal resolutionlarge-depth intravital imaging

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Area of Science:

  • Life Science Research
  • Biomedical Imaging
  • Neuroscience

Background:

  • Dynamic microscopic imaging in vivo is vital for life science.
  • Current limitations include trade-offs between spatial and temporal resolution due to optical throughput.
  • Achieving high resolution at depth in opaque tissues remains challenging.

Purpose of the Study:

  • To develop an intravital microscopy technique with high optical throughput.
  • To overcome the limitations of current microscopy for in vivo dynamic imaging.
  • To enable high-resolution, high-speed imaging of deep tissues in live subjects.

Main Methods:

  • Constructed a near-infrared-II (NIR-II) wide-field fluorescence microscope.
  • Employed a scale-recurrent network to integrate information from two-photon fluorescence microscopy.
  • Utilized deep learning to enhance image quality and resolution.

Main Results:

  • Achieved significant improvements in imaging resolution: five-fold axial and thirteen-fold lateral.
  • Maintained high temporal resolution and light utilization efficiency.
  • Successfully reconstructed 3D vasculature in rodent brains.
  • Observed subtle cerebral vessel changes in models of acute respiratory failure.

Conclusions:

  • The upgraded NIR-II microscope offers high optical throughput for in vivo dynamic imaging.
  • This technology significantly enhances resolution without compromising imaging speed or light efficiency.
  • Enables novel insights into microvascular dynamics in disease states.