Method for automatically detecting spots on a substrate
An apparatus and method for imaging metallic nanoparticles is provided. Preferably, the invention provides for an apparatus and method for detection of gold colloid particles and for accurate reporting to the operator. The apparatus includes a substrate holder for holding the substrate, a processor and memory device, an imaging module, an illumination module, a power module, an input module, and an output module. The apparatus may have a stationary substrate holder and imaging module which are proximate to one another. The apparatus provided for a compact sized system without the need for complex motorized devices to move the camera across the substrate. Further, the apparatus and method provide for automatic detection of the spots/wells on the substrate, automatic quantification of the spots on the substrate, and automatic interpretation of the spots based on decision statistics.
1 - 50 . (canceled)
51 . In a substrate having a plurality of spots containing specific binding complements to one or more target analytes, one of the spots is a first test spot for metallic nanoparticles complexed thereto in the presence of at least a first target analyte, another one of the spots is a second test spot for metallic nanoparticles complexed thereto in the presence of at least a second target analyte, a method of automatically detecting at least some of the plurality of spots, the method comprising:
acquiring at least one image of the plurality of spots composed of metallic nanoparticles on a surface of the substrate;
compensating for at least one type of distortion in the at least one acquired image; and
automatically detecting at least some of the plurality of spots composed of metallic nanoparticles based on the at least one compensated acquired image.
52 . The method of claim 51 , wherein the function of acquiring the at least one image is performed by an image module located at a location less than or equal to 356 mm from the surface of the substrate.
53 . The method of claim 52 , wherein the at least one image acquired by the image module includes all or substantially all of the surface of the substrate.
54 . The method of claim 52 , wherein the image module is a photo sensor.
55 . The method of claim 54 , wherein the photosensor is stationary.
56 . The method of claim 51 ,
wherein the at least one image is acquired using an image device; and
wherein acquiring the at least one image comprises acquiring the image without moving the image device and the substrate relative to one another.
57 . The method of claim 51 , wherein acquiring the at least one image comprises acquiring a plurality of images so as to obtain an optimal image.
58 . The method of claim 51 , wherein compensating for the at least one type of distortion comprises correcting a grayscale distortion in the acquired image.
59 . The method of claim 58 ,
wherein acquiring the at least one image comprises using an image device with a field of view; and
wherein correcting the grayscale distortion comprises applying a compensation model for brightness across the field of view of the image device.
60 . The method of claim 59 , wherein the compensation model is derived by acquiring images using a consistent light source at different brightness values and by using a calibrated set of filters to generate curves for the images acquired at the different brightness values.
61 . The method of claim 51 , wherein compensating for the at least one type of distortion comprises correcting spatial distortion in the acquired image.
62 . The method of claim 61 , wherein correcting spatial distortion in the acquired image comprises:
generating a plurality of points distorted by the spatial distortion;
generating a plurality of points undistorted by spatial distortion;
generating a model based on the plurality of distorted and undistorted points; and
applying the model to the acquired image.
63 . The method of claim 51 , further comprising:
performing adaptive thresholding on at least a portion of the acquired image.
64 . The method of claim 51 , further comprising:
performing a threshold analysis on at least a portion of the acquired image so as to distinguish background areas of the acquired image from foreground areas of the acquired image; and
comparing characteristics of the foreground areas of the acquired image to characteristics of a predetermined spot area,
wherein the foreground areas having characteristics comparable to the characteristics of the predetermined spot area are deemed to be a spot.
65 . The method of claim 64 ,
wherein the characteristics of the foreground areas are selected from the group consisting of area, mass, shape, and circumference, and
wherein the characteristics of the predetermined spot area are selected from the group consisting of area, mass, shape, and circumference.
66 . In a substrate having a plurality of spots containing specific binding complements to one or more target analytes, a method for automatically detecting at least some of the plurality of spots, the method comprising:
acquiring at least one image of the plurality of spots composed of metallic nanoparticles on a surface of the substrate;
compensating for at least one type of distortion in the at least one acquired image;
performing a threshold analysis on at least a portion of the acquired image so as to distinguish background areas of the acquired image from foreground areas of the acquired image;
comparing characteristics of the foreground areas of the acquired image to characteristics of a predetermined spot area, and
automatically detecting at least some of the plurality of spots composed of metallic nanoparticles based on the compensated acquired image,
wherein the foreground areas having characteristics comparable to the characteristics of the predetermined spot area are deemed to be a spot.
67 . The method of claim 66 , wherein compensating for the at least one type of distortion comprises correcting spatial distortion in the acquired image.
68 . The method of claim 66 , wherein compensating for the at least one type of distortion comprises correcting a grayscale distortion in the acquired image.
69 . The method of claim 66 , wherein compensating for the at least one type of distortion comprises correcting a grayscale distortion in the acquired image and correcting spatial distortion in the acquired image.