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Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development
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Multicolor fluorescence microscopy using static light sheets and a single-channel detection.

Jacob Licea-Rodriguez1,2, Alfredo Figueroa-Melendez3, Konstantinos Falaggis2,4

  • 1Centro de Investigación Científica y de Educación Superior de Ensenada, Department of Optics, Ensena, Mexico.

Journal of Biomedical Optics
|January 7, 2019
PubMed
Summary

We developed a cost-effective multicolor fluorescence microscope using selective plane illumination microscopy (SPIM) to image multiple biomarkers simultaneously in living cells. This system enables rapid visualization of cellular dynamics, offering an accessible alternative for biological research.

Keywords:
fluorescence microscopylaser beam combiningmedical and biological imagingmultiple imaging

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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Selective Plane Illumination Microscopy (SPIM) offers optical sectioning capabilities for live biological samples.
  • Multicolor imaging is crucial for visualizing multiple cellular components and their interactions simultaneously.
  • Existing multicolor SPIM systems can be complex and expensive.

Purpose of the Study:

  • To develop a cost-effective, noncommercial multicolor fluorescence microscope system.
  • To enable rapid, simultaneous acquisition of multifluorescent images using a single-camera SPIM setup.
  • To demonstrate the system's capability for visualizing cellular dynamics in living organisms.

Main Methods:

  • Utilized a selective plane illumination microscopy (SPIM) configuration.
  • Employed three continuous wave-lasers modulated with time-delayed pulse trains.
  • Synchronized laser excitation at one-third of the camera frame rate for sequential biomarker excitation and image acquisition.
  • Used a single-channel-detection camera.

Main Results:

  • Successfully demonstrated multicolor imaging of living hyphae of Neurospora crassa.
  • Acquired single-plane multicolor images, visualizing localization and dynamics of multiple cellular components.
  • Validated the system for simultaneous visualization of different cellular processes in living hyphae.

Conclusions:

  • The presented SPIM configuration provides a cost-effective alternative for rapid, simultaneous multifluorescent image acquisition.
  • This system facilitates the study of cellular localization and dynamics of multiple biomarkers in living samples.
  • The technology holds potential for three-dimensional imaging of large biological specimens.