Surface plasmon resonance imaging of cells and surface-associated fibronectin.
BACKGROUND
A critical challenge in cell biology is quantifying the interactions of cells with their extracellular matrix (ECM) environment and the active remodeling by cells of their ECM. Fluorescence microscopy is a commonly employed technique for examining cell-matrix interactions. A label-free imaging method would provide an alternative that would eliminate the requirement of transfected cells and modified biological molecules, and if collected nondestructively, would allow long term observation and analysis of live cells.


RESULTS
Using surface plasmon resonance imaging (SPRI), the deposition of protein by vascular smooth muscle cells (vSMC) cultured on fibronectin was quantified as a function of cell density and distance from the cell periphery. We observed that as much as 120 ng/cm2 of protein was deposited by cells in 24 h.


CONCLUSIONS
SPRI is a real-time, low-light-level, label-free imaging technique that allows the simultaneous observation and quantification of protein layers and cellular features. This technique is compatible with live cells such that it is possible to monitor cellular modifications to the extracellular matrix in real-time.
Peterson
Alexander W
AW
Cell and Tissue Measurements Group, Biochemical Sciences Division, National Institute of Standards and Technology, Gaithersburg, MD, USA. alexander.peterson@nist.gov
Halter
Michael
M
Tona
Alessandro
A
Bhadriraju
Kiran
K
Plant
Anne L
AL
D000818:Animals; D002460:Cell Line; D002477:Cells / Q000648:ultrastructure; D003513:Cycloheximide / Q000494:pharmacology; D005109:Extracellular Matrix / Q000648:ultrastructure*; D005353:Fibronectins / Q000378:metabolism* / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism*; D051381:Rats; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*
D005353:Fibronectins; D003513:Cycloheximide