Limits...
Intensity interrogation near cutoff resonance for label-free cellular profiling.

Nazirizadeh Y, Behrends V, Prósz A, Orgovan N, Horvath R, Ferrie AM, Fang Y, Selhuber-Unkel C, Gerken M - Sci Rep (2016)

Bottom Line: We report a method enabling intensity-based readout for label-free cellular assays, and realize a reader device with the same footprint as a microtiter plate.For unambiguous resonance intensity measurements in resonance waveguide grating (RWG) sensors, we propose to apply resonances near the substrate cutoff wavelength.The significantly reduced size of the reader device opens new opportunities for easy integration into incubators or liquid handling systems.

View Article: PubMed Central - PubMed

Affiliation: Byosens GmbH, 20357 Hamburg, Germany.

ABSTRACT
We report a method enabling intensity-based readout for label-free cellular assays, and realize a reader device with the same footprint as a microtiter plate. For unambiguous resonance intensity measurements in resonance waveguide grating (RWG) sensors, we propose to apply resonances near the substrate cutoff wavelength. This method was validated in bulk refractive index, surface bilayer and G protein-coupled receptor (GPCR) experiments. The significantly reduced size of the reader device opens new opportunities for easy integration into incubators or liquid handling systems.

No MeSH data available.


Related in: MedlinePlus

Resonant wavelength grating (RWG) sensor for cellular assays.(a) Illustration of the RWG sensor with a surface coating. The surface mass of cells grown on the surface is detected within the penetration depth. The interrogation of the RWG sensor in a spectral reflection measurement. (b) When compounds are added to the well, two main cellular responses are detected: (1) mass change and (2) dynamic mass redistribution (DMR). (c) Reflection spectrum of the RWG sensor under 26°. Blue and red arrows indicate the resonance at near-cutoff and non-cutoff wavelengths, respectively. The inset shows the appearing resonance mode curve close to the substrate cutoff for the TM1 mode. (d) Cell confluency in an adhesion and spreading assay. (e) Resonance central wavelength, (f) width change and (g) intensity change in percent during the adhesion and spreading assay. (h) The comparison of the central wavelength of the non-cutoff resonance and the intensity of the near-cutoff resonance shows that the intensity of the near-cutoff resonance can be utilized for surface mass detection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4834563&req=5

f1: Resonant wavelength grating (RWG) sensor for cellular assays.(a) Illustration of the RWG sensor with a surface coating. The surface mass of cells grown on the surface is detected within the penetration depth. The interrogation of the RWG sensor in a spectral reflection measurement. (b) When compounds are added to the well, two main cellular responses are detected: (1) mass change and (2) dynamic mass redistribution (DMR). (c) Reflection spectrum of the RWG sensor under 26°. Blue and red arrows indicate the resonance at near-cutoff and non-cutoff wavelengths, respectively. The inset shows the appearing resonance mode curve close to the substrate cutoff for the TM1 mode. (d) Cell confluency in an adhesion and spreading assay. (e) Resonance central wavelength, (f) width change and (g) intensity change in percent during the adhesion and spreading assay. (h) The comparison of the central wavelength of the non-cutoff resonance and the intensity of the near-cutoff resonance shows that the intensity of the near-cutoff resonance can be utilized for surface mass detection.

Mentions: A RWG sensor utilizes a periodically nanostructured waveguide to couple the incident light into the waveguide, and provide a leaky mode with an evanescent part, which is measured as a resonance in a reflection interrogation. The evanescent part of the mode typically exhibits a penetration depth of about 200 nanometers above the sensor structure (Fig. 1a), which is the sensing volume of the RWG sensor. Here we used Corning 96-well Epic microtiter plates with a RWG sensor integrated in each well.


Intensity interrogation near cutoff resonance for label-free cellular profiling.

Nazirizadeh Y, Behrends V, Prósz A, Orgovan N, Horvath R, Ferrie AM, Fang Y, Selhuber-Unkel C, Gerken M - Sci Rep (2016)

Resonant wavelength grating (RWG) sensor for cellular assays.(a) Illustration of the RWG sensor with a surface coating. The surface mass of cells grown on the surface is detected within the penetration depth. The interrogation of the RWG sensor in a spectral reflection measurement. (b) When compounds are added to the well, two main cellular responses are detected: (1) mass change and (2) dynamic mass redistribution (DMR). (c) Reflection spectrum of the RWG sensor under 26°. Blue and red arrows indicate the resonance at near-cutoff and non-cutoff wavelengths, respectively. The inset shows the appearing resonance mode curve close to the substrate cutoff for the TM1 mode. (d) Cell confluency in an adhesion and spreading assay. (e) Resonance central wavelength, (f) width change and (g) intensity change in percent during the adhesion and spreading assay. (h) The comparison of the central wavelength of the non-cutoff resonance and the intensity of the near-cutoff resonance shows that the intensity of the near-cutoff resonance can be utilized for surface mass detection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834563&req=5

f1: Resonant wavelength grating (RWG) sensor for cellular assays.(a) Illustration of the RWG sensor with a surface coating. The surface mass of cells grown on the surface is detected within the penetration depth. The interrogation of the RWG sensor in a spectral reflection measurement. (b) When compounds are added to the well, two main cellular responses are detected: (1) mass change and (2) dynamic mass redistribution (DMR). (c) Reflection spectrum of the RWG sensor under 26°. Blue and red arrows indicate the resonance at near-cutoff and non-cutoff wavelengths, respectively. The inset shows the appearing resonance mode curve close to the substrate cutoff for the TM1 mode. (d) Cell confluency in an adhesion and spreading assay. (e) Resonance central wavelength, (f) width change and (g) intensity change in percent during the adhesion and spreading assay. (h) The comparison of the central wavelength of the non-cutoff resonance and the intensity of the near-cutoff resonance shows that the intensity of the near-cutoff resonance can be utilized for surface mass detection.
Mentions: A RWG sensor utilizes a periodically nanostructured waveguide to couple the incident light into the waveguide, and provide a leaky mode with an evanescent part, which is measured as a resonance in a reflection interrogation. The evanescent part of the mode typically exhibits a penetration depth of about 200 nanometers above the sensor structure (Fig. 1a), which is the sensing volume of the RWG sensor. Here we used Corning 96-well Epic microtiter plates with a RWG sensor integrated in each well.

Bottom Line: We report a method enabling intensity-based readout for label-free cellular assays, and realize a reader device with the same footprint as a microtiter plate.For unambiguous resonance intensity measurements in resonance waveguide grating (RWG) sensors, we propose to apply resonances near the substrate cutoff wavelength.The significantly reduced size of the reader device opens new opportunities for easy integration into incubators or liquid handling systems.

View Article: PubMed Central - PubMed

Affiliation: Byosens GmbH, 20357 Hamburg, Germany.

ABSTRACT
We report a method enabling intensity-based readout for label-free cellular assays, and realize a reader device with the same footprint as a microtiter plate. For unambiguous resonance intensity measurements in resonance waveguide grating (RWG) sensors, we propose to apply resonances near the substrate cutoff wavelength. This method was validated in bulk refractive index, surface bilayer and G protein-coupled receptor (GPCR) experiments. The significantly reduced size of the reader device opens new opportunities for easy integration into incubators or liquid handling systems.

No MeSH data available.


Related in: MedlinePlus