GRATING BASED SENSOR COMBINING LABEL-FREE BINDING DETECTION AND FLUORESNCE AMPLIFICATION AND READOUT SYSTEM FOR SENSOR
First Claim
1. A method of conducting a cell-based assay of a sample comprising one or more cells, the method comprising the steps of:
- (a) providing a grating-based biosensor substrate having a periodic surface grating structure wherein the periodic grating structure is constructed in a manner designed for both
1) optical interrogation of the sensor with light in an evanescent resonance (ER) detection mode, and
2) optical interrogation of the sensor with light in a label-free detection mode;
(b) applying the sample to the biosensor;
(c) using the biosensor to measure at least one of following;
(a) cell attachment of cells in the sample to the surface of the grating structure, (b) adhesion of cells in the sample to the sensor or to an extracellular matrix coating the sensor, (c) adhesion of cells in the sample to other cells, (d) morphological change of cells in the sample, (e) chemotaxis of cells in the sample, (f) protein exocytosis from the cell into the vicinity of the sensor, (g) ion flux in and out of the cell in the vicinity of the sensor, (h) cell migration, (i) cell flattening or rounding, (j) cell growth or differentiation, (k) cell death, (l) cytoskeletal rearrangement of cells in the sample, and (m) rearrangement, reorganization or changed expression of proteins inside or outside of the cells in the sample;
and(d) using the biosensor to measure the effect of at least one of the following on cell function of cells in the sample;
(a) a drug compound candidate, (b) a protein, (c) a peptide or modified peptide, (d) an antibody or fragment thereof, (e) DNA including modified DNA (known in the art for eliciting biological effects), (f) RNAi including modified RNAi, (g) RNA (including modified RNA), (h) a chemokine, (i) a virus, (j) other cells, including bacteria or other organisms, (k) engineered binding domains (e.g., Affibodies, DARPins, Adnectins), (l) sugars or modified sugars, (m) ions, (n) lipids and modified lipids, (o) metals, (p) inorganic solvents, and (q) organic solvents
2 Assignments
0 Petitions
Accused Products
Abstract
A grating-based sensor is disclosed that has a grating structure constructed and designed for both evanescent resonance (ER) fluorescence detection and label-free detection applications. One and two-dimensional gratings are also disclosed, including gratings characterized by unit cells with central posts, central holes, and two-level, two-dimensional gratings. A readout system for such sensors is also disclosed. Various applications for the biosensors are described, including cell-based assays for assessing the effect of drug compounds, proteins, peptides and other materials on cell function. A biosensor embodiment optimized for a luminescent response at two different wavelengths is also described. Such luminescent response could be produced by fluorescence (either native or from an attached fluorophore), phosphorescence, chemi-luminescence, or other luminescence technology. Two different luminescence technologies could be combined on the same biosensor chip.
74 Citations
44 Claims
-
1. A method of conducting a cell-based assay of a sample comprising one or more cells, the method comprising the steps of:
-
(a) providing a grating-based biosensor substrate having a periodic surface grating structure wherein the periodic grating structure is constructed in a manner designed for both
1) optical interrogation of the sensor with light in an evanescent resonance (ER) detection mode, and
2) optical interrogation of the sensor with light in a label-free detection mode;(b) applying the sample to the biosensor; (c) using the biosensor to measure at least one of following;
(a) cell attachment of cells in the sample to the surface of the grating structure, (b) adhesion of cells in the sample to the sensor or to an extracellular matrix coating the sensor, (c) adhesion of cells in the sample to other cells, (d) morphological change of cells in the sample, (e) chemotaxis of cells in the sample, (f) protein exocytosis from the cell into the vicinity of the sensor, (g) ion flux in and out of the cell in the vicinity of the sensor, (h) cell migration, (i) cell flattening or rounding, (j) cell growth or differentiation, (k) cell death, (l) cytoskeletal rearrangement of cells in the sample, and (m) rearrangement, reorganization or changed expression of proteins inside or outside of the cells in the sample;and (d) using the biosensor to measure the effect of at least one of the following on cell function of cells in the sample;
(a) a drug compound candidate, (b) a protein, (c) a peptide or modified peptide, (d) an antibody or fragment thereof, (e) DNA including modified DNA (known in the art for eliciting biological effects), (f) RNAi including modified RNAi, (g) RNA (including modified RNA), (h) a chemokine, (i) a virus, (j) other cells, including bacteria or other organisms, (k) engineered binding domains (e.g., Affibodies, DARPins, Adnectins), (l) sugars or modified sugars, (m) ions, (n) lipids and modified lipids, (o) metals, (p) inorganic solvents, and (q) organic solvents- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 40, 41, 42)
-
- 16. A grating-based biosensor comprising a substrate having a periodic surface grating structure wherein the periodic grating structure is constructed in a manner designed for optical interrogation of the sensor with excitation light at least two discrete wavelengths in an evanescent resonance (ER) detection mode to thereby produce a luminescence response from a sample placed on the biosensor at least two discrete emission wavelengths.
- 24. A grating-based biosensor comprising a substrate having a periodic surface grating structure wherein the periodic grating structure is constructed in a manner designed for optical interrogation of the sensor with light in an evanescent resonance (ER) detection mode to produce a luminescence response from a sample placed on the biosensor at two discrete wavelengths, and wherein the luminescence response at the two discrete wavelengths is produced from two different types of luminescence.
-
31. A method of conducting a cell-based assay of a sample comprising one or more cells, the method comprising the steps of:
-
providing a grating-based biosensor substrate having a periodic surface grating structure wherein the periodic grating structure is constructed in a manner designed for both
1) optical interrogation of the sensor with light in an evanescent resonance (ER) detection mode, and
2) optical interrogation of the sensor with light in a label-free detection mode;applying a sample to the cell, using the biosensor to measure cell attachment of cells in the sample to the surface of the grating structure, and using the biosensor to measure the effect of a drug compound candidate on cell function of the cells in the sample. - View Dependent Claims (32, 33, 34, 35, 36, 37, 38, 39)
-
-
43. A method of conducting a cell-based assay of a sample comprising one or more cells, the method comprising the steps of:
-
(a) providing a grating-based biosensor substrate having a periodic surface grating structure wherein the periodic grating structure is constructed in a manner designed for both
1) optical interrogation of the sensor with light in an evanescent resonance (ER) detection mode, and
2) optical interrogation of the sensor with light in a label-free detection mode;(b) applying the sample to the biosensor; and (c) monitoring interactions of at least two different cell types wherein one cell type is measured in the label-free and another cell type monitored in the ER mode. - View Dependent Claims (44)
-
Specification