Analysis of somitogenesis using multiphoton laser scanning microscopy (MPLSM)

Mary E. Dickinson; Kenneth J. Longmuir; Scott E. Fraser

doi:10.1117/12.424570

24 April 2001 Analysis of somitogenesis using multiphoton laser scanning microscopy (MPLSM)

Mary E. Dickinson, Kenneth J. Longmuir, Scott E. Fraser

Proceedings Volume 4262, Multiphoton Microscopy in the Biomedical Sciences; (2001) https://doi.org/10.1117/12.424570
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

In order to study complex cellular interactions in the developing somite and nervous system, we have been refining techniques for labeling and imaging individual cells within the living vertebrate embryo. Most recently, we have been using MPLSM to analyze cellular behaviors, such as cell migration, filopodial extension, cell process collapse, and neuron pathfinding using time-lapse microscopy in 3-dimensions (3-d). To enhance the efficiency of two-photon excitation in these samples, we have been using a Zeiss LSM 510 NLO fiber delivery system with a Grating Dispersion Compensator (GDC). This system not only offers the convenience of fiber delivery for coupling our Ti:Sapphire laser to the microscope, but also affords us precise control over the pulsewidth of the mode- locked beam. In addition, we have developed a novel peptide/non-cationic lipid gene delivery system to introduce GFP plasmid into somite cells. This approach has allowed us to generate detailed 3-d images of somite cell morphologies at various stages of somite development in a way that best preserves the vitality of the cells being imaged.

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Mary E. Dickinson, Kenneth J. Longmuir, and Scott E. Fraser "Analysis of somitogenesis using multiphoton laser scanning microscopy (MPLSM)", Proc. SPIE 4262, Multiphoton Microscopy in the Biomedical Sciences, (24 April 2001); https://doi.org/10.1117/12.424570

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KEYWORDS

Green fluorescent protein

Multiphoton microscopy

Microscopes

Tissues

3D image processing

Laser scanners

Microscopy

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