Embryonic lineage analysis using three-dimensional, time-lapse in-vivo fluorescent microscopy

Jonathan Minden; Zvi Kam; David A. Agard; John W. Sedat; Bruce Alberts

doi:10.1117/12.17781

1 August 1990 Embryonic lineage analysis using three-dimensional, time-lapse in-vivo fluorescent microscopy

Jonathan Minden, Zvi Kam, David A. Agard, John W. Sedat, Bruce Alberts

Proceedings Volume 1205, Bioimaging and Two-Dimensional Spectroscopy; (1990) https://doi.org/10.1117/12.17781
Event: OE/LASE '90, 1990, Los Angeles, CA, United States

Abstract

Drosophila melanogaster has become one of the most extensively studied organisms because of its amenability to genetic analysis. Unfortunately, the biochemistry and cell biology ofDrosophila has lagged behind. To this end we have been microinjecting fluorescently labelled proteins into the living embryo and observing the behavior of these proteins to determine their role in the cell cycle and development. Imaging of these fluorescent probes is an extremely important element to this form of analysis. We have taken advantage of the sensitivity and well behaved characteristics of the charge coupled device (CCD) camera in conjunction with digital image enhancement schemes to produce highly accurate images of these fluorescent probes in vivo. One of our major goals is to produce a detailed map of cell fate so that we can understand how fate is determined and maintained. In order produce such a detailed map, protocols for following the movements and mitotic behavior of a large number of cells in three dimensions over relatively long periods of time were developed. We will present our results using fluorescently labelled histone proteins as a marker for nuclear location1. In addition, we will also present our initial results using a photoactivatable analog of fluorescein to mark single cells so that their long range fate can be unambiguously determined.

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Jonathan Minden, Zvi Kam, David A. Agard, John W. Sedat, and Bruce Alberts "Embryonic lineage analysis using three-dimensional, time-lapse in-vivo fluorescent microscopy", Proc. SPIE 1205, Bioimaging and Two-Dimensional Spectroscopy, (1 August 1990); https://doi.org/10.1117/12.17781

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KEYWORDS

Proteins

Spectroscopy

In vivo imaging

3D image processing

CCD cameras

Biological research

Image enhancement

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