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Related Concept Videos

Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

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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

Updated: Mar 28, 2026

Three and Four-Dimensional Visualization and Analysis Approaches to Study Vertebrate Axial Elongation and Segmentation
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A virtually imaged defocused array (VIDA) for high-speed 3D microscopy.

Ethan Schonbrun1, Giuseppe Di Caprio2

  • 1Rowland Institute at Harvard, 100 Edwin H. Land Blvd., Cambridge, MA 02142, USA. ethanschonbrun@gmail.com.

Journal of Biophotonics
|December 24, 2015
PubMed
Summary
This summary is machine-generated.

We developed a fast 3D imaging method using a custom etalon to capture multifocus image stacks in a single exposure. This technique resolves neutrophil nuclear morphology in microfluidic flow and enables 3D imaging flow cytometry.

Keywords:
Cytometrymicrofluidicsmicroscopy

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Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • Traditional microscopy struggles with capturing dynamic 3D cellular structures in high-throughput applications.
  • Resolving the complex, dynamic 3D morphology of nuclei, especially during cellular processes like deformation, requires advanced imaging techniques.

Purpose of the Study:

  • To introduce a novel method for rapid multifocus image stack acquisition using a custom etalon.
  • To demonstrate the application of this technique for analyzing the 3D morphology of neutrophil nuclei in microfluidic flow.
  • To develop a 3D imaging flow cytometer for high-velocity cellular analysis.

Main Methods:

  • A custom etalon was employed to generate multiple reflections, creating a multifocus image stack captured in a single, short camera exposure (100 µs).
  • The VIDA microscope, incorporating this etalon-based method, was utilized for imaging.
  • A 3D imaging flow cytometer was constructed and used to analyze white blood cells.

Main Results:

  • The method successfully captured multifocus image stacks, enabling 3D reconstruction with minimal camera integration time.
  • The 3D morphology of neutrophil nuclei was resolved as they rotated and deformed within a microfluidic constriction.
  • Nearly a thousand white blood cells flowing at 3 mm/s had their nuclear morphology quantified using the 3D imaging flow cytometer.

Conclusions:

  • The developed etalon-based multifocus imaging technique provides a compact, simple, and versatile solution for rapid 3D cellular imaging.
  • This method is suitable for high-throughput analysis, such as in 3D imaging flow cytometry, and for studying dynamic cellular processes.
  • The VIDA microscope offers advantages like intrinsic achromatism and easy reconfigurability of optical parameters.