Measuring diffusion coefficients of labelled particles on cell surfaces by digital fluorescence microscopy

1990 ◽  
Vol 18 (5) ◽  
pp. 938-938 ◽  
Author(s):  
IAN E. G. MORRISON ◽  
CATHERINE M. ANDERSON ◽  
GEORGE N. GEORGIOU ◽  
RICHARD J. CHERRY
Keyword(s):  
Fluorescence Microscopy ◽  
Diffusion Coefficients ◽  
Cell Surfaces ◽  
Digital Fluorescence Microscopy

Trimers of the fibronectin cell adhesion domain localize to actin filament bundles and undergo rearward translocation

2002 ◽  
Vol 115 (12) ◽  
pp. 2581-2590 ◽  
Author(s):  
Françoise Coussen ◽  
Daniel Choquet ◽  
Michael P. Sheetz ◽  
Harold P. Erickson
Keyword(s):  
Cell Adhesion ◽  
Fluorescence Microscopy ◽  
Cell Surface ◽  
Actin Cytoskeleton ◽  
Fiber Bundles ◽  
Cell Surfaces ◽  
Actin Fiber ◽  
Filament Bundles ◽  
Necessary And Sufficient ◽  
Small Clusters

Previous studies have shown that small beads coated with FN7-10, a four-domain cell adhesion fragment of fibronectin, bind to cell surfaces and translocate rearward. Here we investigate whether soluble constructs containing two to five FN7-10 units might be sufficient for activity. We have produced a monomer, three forms of dimers, a trimer and a pentamer of FN7-10,on the end of spacer arms. These oligomers could bind small clusters of up to five integrins. Fluorescence microscopy showed that the trimer and pentamer bound strongly to the cell surface, and within 5 minutes were prominently localized to actin fiber bundles. Monomers and dimers showed only diffuse localization. Beads coated with a low concentration (probably one complex per bead) of trimer or pentamer showed prolonged binding and rearward translocation, presumably with the translocating actin cytskeleton. Beads containing monomer or dimer showed only brief binding and diffusive movements. We conclude that clusters of three integrin-binding ligands are necessary and sufficient for coupling to and translocating with the actin cytoskeleton.

scholarly journals Automated high-throughput digital fluorescence microscopy for TB diagnosis

2020 ◽  
Vol 24 (10) ◽  
pp. 1103-1105
Author(s):  
D. Chesov ◽  
V. Lesanu ◽  
N. Ciobanu ◽  
A. Codreanu ◽  
V. Crudu ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
High Throughput ◽  
Digital Fluorescence Microscopy

Visualization of replication sites in unfixed human cells

1993 ◽  
Vol 105 (2) ◽  
pp. 541-550 ◽  
Author(s):  
A.B. Hassan ◽  
P.R. Cook
Keyword(s):  
Dna Replication ◽  
Fluorescence Microscopy ◽  
Hela Cells ◽  
Human Cells ◽  
Underlying Structure ◽  
Fixation Methods ◽  
Replication Sites ◽  
Digital Fluorescence Microscopy ◽  
Replication Activity

Sites of DNA replication in nuclei are focally concentrated, suggesting that an underlying structure organizes the activity of many polymerases. As fixation could induce aggregation into foci, we examined the distribution of replication sites in unfixed nuclei. HeLa cells were encapsulated in agarose microbeads, permeabilized in a ‘physiological’ buffer, their DNA polymerizing activity characterized, and replication sites directly labelled by incubation with fluorochrome-dUTP conjugates. Using conventional and digital fluorescence microscopy, 80–250 foci were seen in these unfixed cells. These foci are unlikely to be formed by the aggregation of separate polymerases as most replication activity found in vivo is retained throughout these procedures. Although commonly used fixation methods collapsed or dispersed their periphery, the central core was very stable. Foci remained when approximately 90% chromatin was removed, suggesting they were attached to an underlying structure.

Epi-fluorescence Microscopy and Image Analysis Used to Measure Diffusion Coefficients in Gel Systems

1992 ◽  
Vol 44 (7) ◽  
pp. 543-549 ◽  
Author(s):  
B. T. HENRY ◽  
J. ADLER ◽  
S. HIBBERD ◽  
M. S. CHEEMA ◽  
S. S. DAVIS ◽  
...  
Keyword(s):  
Image Analysis ◽  
Fluorescence Microscopy ◽  
Diffusion Coefficients
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