Biology Reference
In-Depth Information
10.3.5 Strengths and Limitaons of AFM-Based SCFS
With the described AFM-based SCFS setup, various cell adhesion experiments
can be performed under near-physiological conditions. AFM-based SCFS allows
to measure adhesive forces that range from a few piconewtons up to several
hundred nanonewtons. Thus, interactions mediated by single CAMs
28,29,35,46,64
or adhesive interactions established by larger adhesive complexes can
be detected in the cellular context.
41,60
Although other SCFS methods can
provide a better force resolution (optical tweezer, BFP), AFM-based SCFS
is more versatile in terms of the detectable range of adhesive forces. This
advantage makes it possible to address a broad range of biological questions.
Another advantage over other SCFS assays is the high precision with which
the cell can be temporally and spatially manipulated. AFM-based SCFS can
be easily combined with optical techniques such as total internal relection
luorescence microscopy, confocal microscopy, luorescent microscopy and
conventional transmission light microscopy. AFMs speciically developed to
perform SCFS are commercially available and relatively easy to use. In the
future, establishing cellular assays and standardized experimental protocols
for SCFS together with automated data analysis tools will enable newcomers
and professionals to explore the molecular mechanisms of cell adhesion.
References
1.
Adams, J. C., and Watt F. M. (1993) Regulation of development and differentiation
by the extracellular matrix,
Development
,
117
, 1183-1198.
2.
Bissell, M. J., Hall, H. G., and Parry, G. (1982) How does the extracellular matrix
direct gene expression?
J. Theor. Biol.
,
99
, 31-68.
3.
Cukierman, E., Pankov, R., and Yamada, K. M. (2002) Cell interactions with
three-dimensional matrices,
Curr. Opin. Cell Biol.
,
14
, 633-639.
4. Damsky, C., Sutherland, A., and Fisher, S. (1993) Extracellular matrix 5:
adhesive interactions in early mammalian embryogenesis, implantation, and
placentation,
FASEB J.
,
7
, 1320-1329.
5.
Damsky, C. H., and Ilic, D. (2002) Integrin signalling: it´s where the action is,
Curr. Opin. Cell Biol.
,
14
, 594-602.
6.
Juliano, R. L., and Haskill, S. (1993) Signal transduction from the extracellular
matrix,
J. Cell Biol.
,
120
, 577-585.
7.
Meighan, C. M., and Schwarzbauer, J. E. (2008) Temporal and spatial regulation
of integrins during development,
Curr. Opin. Cell Biol.
,
20
, 520-524.
8.
Bell, G. I. (1978) Models for speciic adhesion of cells to cells,
Science
,
200
,
618-627.
9.
Connors, W. L., and Heino, J. (2005) A duplexed microsphere-based cellular
adhesion assay,
Anal. Biochem.
,
337
, 246-255.
Search WWH ::
Custom Search