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FLUID-CELL: Flow-enabled Light and Ultrastructural Imaging Device for Correlative Electron and Light Localization.

Nicholas M Rienstra1,2, Steve Garvis1,3,4, Juan C Sanchez1

  • 1Department of Biochemistry, University of Wisconsin, Madison, WI, USA.

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A new microfluidic flow cell, the FLUID-CELL, links fluorescence light microscopy (FLM) and cryo-electron microscopy (cryo-EM) for advanced biological imaging. This device enables correlative ultrastructural and functional analyses of cells at the nanoscale.

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bacteriabiofilmcryo-electron microscopy (cryo-EM)cryo-electron tomography (cryo-ET)microfluidicthree-dimensional (3D) printing

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Area of Science:

  • Cell Biology
  • Microscopy
  • Nanotechnology

Background:

  • Correlative light and electron microscopy (CLEM) is crucial for understanding cellular ultrastructure and function.
  • Existing CLEM techniques can be limited by sample preparation and workflow complexity.

Purpose of the Study:

  • To introduce a novel microfluidic flow cell, FLUID-CELL, for seamless integration of fluorescence light microscopy (FLM) and cryo-electron microscopy (cryo-EM).
  • To enable correlative imaging of dynamic cellular events with high-resolution ultrastructural data.

Main Methods:

  • Development of a microfluidic flow cell (FLUID-CELL) with a precisely engineered microchannel.
  • Integration of FLM for real-time observation and cryo-EM for ultrastructural analysis within the same device.
  • Maintaining native cell culturing conditions throughout the imaging process.

Main Results:

  • Demonstrated practical FLM imaging over extended periods.
  • Achieved consistent sample handling and enabled correlative imaging.
  • Successfully connected dynamic cellular processes observed by FLM with high-resolution cryo-EM data.

Conclusions:

  • The FLUID-CELL device streamlines the workflow for dual-modality imaging.
  • This approach opens new avenues for investigating the relationship between cellular function and molecular architecture.
  • Facilitates nanoscale structural and ultrastructural analyses of biological samples.