F-actin organization during the cellularization of the Drosophila embryo as revealed with a confocal laser scanning microscope

1990 ◽  
Vol 96 (1) ◽  
pp. 35-42
Author(s):  
R.M. Warn ◽  
M. Robert-Nicoud

The changes in F-actin organization during the cellularization of the Drosophila embryo have been studied with a confocal laser scanning microscope using fluorescein-phalloidin as a specific stain. Particular study has been made of the changes in the organization of the F-actin network associated with the leading edges of the growing membranes. The role of this actin network in the cellularization process is considered. Other actin-containing structures have also been examined, including the cortical actin layer and a conspicuous region of F-actin aggregates, present beneath the level of the forming cell membranes.

Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.


2010 ◽  
Vol 42 (7) ◽  
pp. 662-670 ◽  
Author(s):  
Marcelo Tavares de Oliveira ◽  
Cesar Augusto Galvão Arrais ◽  
Ana Cecília Aranha ◽  
Carlos de Paula Eduardo ◽  
Katsuya Miyake ◽  
...  

1994 ◽  
Vol 266 (1) ◽  
pp. C303-C310 ◽  
Author(s):  
E. Niggli ◽  
D. W. Piston ◽  
M. S. Kirby ◽  
H. Cheng ◽  
D. R. Sandison ◽  
...  

In this paper we describe the modifications necessary to upgrade, at affordable cost, a commercially available confocal laser scanning microscope for use with ultraviolet (UV) excitation. The optical problems associated with these modifications are described in detail, and easy solutions to solve them are suggested. The optical resolution of the instrument was tested with fluorescent beads and was found to be close to diffraction limited. The light losses due to lateral chromatic aberration were assessed in a thick fluorescent specimen and were found to be comparable to those usually observed with visible light. For a more visual example of the resolution of this instrument, isolated ventricular heart muscle cells were loaded with the fluorescent Ca2+ indicator indo 1. This allowed us to visualize subcellular structural detail and to illustrate the optical sectioning capability of the UV confocal microscope when recording indo 1 emission. Dual-emission line scans were used to perform ratiometric time-resolved detection of Ca2+ transients in voltage-clamped heart muscle cells loaded with the salt form of indo 1. The system presented in this paper should significantly broaden the range of fluorescent indicators that can be used in confocal microscopy.


1993 ◽  
Vol 14 (Supplement) ◽  
pp. 407-410
Author(s):  
Chikako Tokuda ◽  
Katsuhiko Tsujioka ◽  
Hiroyuki Tachibana ◽  
Yasuo Ogasawara ◽  
Tokunori Yamamoto ◽  
...  

2007 ◽  
Vol 33 (8) ◽  
pp. 957-961 ◽  
Author(s):  
Saman R. Gharib ◽  
Patricia A. Tordik ◽  
Glen M. Imamura ◽  
Thomas A. Baginski ◽  
Gary G. Goodell

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