OWLS APPLICATION NOTES NO-004

QUANTIFICATION OF CELL ADHESIVITY

using OPTICAL WAVEGUIDE LIGHTMODE SPECTROSCOPY (OWLS) detection

Abstract
The molecular interactions and cell-biological mechanisms behind the adhesive behavior of a cell are not properly understood.
In the last decades, the development of bio-protheses and the hope for elaboration of successful cell replacement therapies
accelerated the search for appropriate scaffold materials and conditions which may help to keep cells alive and functioning on
artificial surfaces.
For understanding some crucial steps of cell-attachment, the initial phases of attachment of two different types of cloned cells –
NE-4C neural stem and MDCK epithelial cells - were analyzed by optical waveguide light mode spectroscopic (OWLS) methods,

Application of OWLS sensors Detection of substrate-dependent alterations

as selective detectors of substrate-near cell components in initial cell attachment
Distance from the surface
Attachment points SiO2/Ti
10
39
0

34 PL
Evanescent light
0

Absorbed mass [arb.units /

29
100 0 Lamini
Waveguide 24

19
0 PLL+F
14
OWLS methods allow quantifying the deposition of material in a 0
thin (<150 nm) layer above the solid sensor surface. In terms of 9
cells, it can provide data on focal contacts and adhesion sites, 4
while the rest of the cellular mass remains out of the field of 0
-
detection. 23 23 24 24 25 25 26 26 27
2 7 2 7 Time
2 7 2 7 2
[min]
Surface chemistry, sensitization of the
The length of the latency period depends on the adhesivity of the
sensor surface provided substrate
Bare or amino-functionalized sensor surfaces were coated with
different attachment-molecules including poly-L-lysine, laminin,
fibronectin, and RGD peptide-mimetics.
Deposition of NE 4C cells on laminin
34
Cell preparations 0
Cell suspensions in protein-free, artificial cerebro-spinal fluid were 29 Active processes
37oC
Absorbed mass [arb.units / area]

introduced into the measuring cuvette, and the cell - substrate 0

interactions were monitored up to 80 – 120 min. For control, the 24
“attachment” of cells fixed with paraformaldehyde, poisoned by 0 latency
Cytochalasin B, or sedimented at +4 oC were also assayed. 19
0
14
300 NE-4C 300 MDCK 0
9 6oC
250 250 0
D e p o s ite d m a te r ia l

D e p o s itie d m a te ri a l

4
200 200 0
[a r b .u n its ]

[a r b .u n its ]

-
0 10 20 30 40 50 60 70
150 150 10
Time [min]
100 100
Conclusion
50 50
The ∆N versus time function clearly distinguished the active
0
attachment from passive sedimentation.
0 The results indicate that OWLS techniques allow rapid evaluation
5 15 25 35 45 55 65
5 15 25 35 45 55 of cell – matrix interactions and provide tools to characterize the
Time [min] Time [min]
composition of sets of adhesion molecules on cell surfaces.

OWLS signals in the first 60 min of attachment References

OWLS recordings revealed characteristic differences in cell- 1. Vörös J. et al., Optical grating coupler biosensors. Biomaterials
behavior between the two types of cells 23 (2002) 3699-3710.
2. Madarasz E. et al., (2006): Attachment of NE-4C
neuroectodermal stem cells to different surfaces: evaluation of
cell – substrate interactions by optical waveguide light mode
spectroscopy (OWLS) FEBS Letts. In press
3. Madarasz E. et al., (2006):Label-free OWLS assays on cell
adhesivity. SBS Conference Seattle
4. www.owls-sensors.com

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