Methods and apparatus for detection of fluorescently labeled materials
View Patent ↗Fluorescently marked targets bind to a substrate 230 synthesized with polymer sequences at known locations. The targets are detected by exposing selected regions of the substrate 230 to light from a light source 100 and detecting the photons from the light fluoresced therefrom, and repeating the steps of exposure and detection until the substrate 230 is completely examined. The resulting data can be used to determine binding affinity of the targets to specific polymer sequences.
1. A scanning system, comprising:
a probe array;
one or more optical elements constructed and arranged to direct an excitation beam at the probe array;
one or more detectors constructed and arranged to receive reflected intensity data responsive to the excitation beam, wherein the reflected intensity data is responsive, at least in part, to a focusing distance between an optical element and the probe array;
a transport frame constructed and arranged to adjust the focusing distance in a first direction with respect to the probe array;
an auto-focuser constructed and arranged to determine a best plane of focus based, at least in part, upon one or more characteristics of the reflected intensity data as received at two or more focusing distances;
and wherein:
the one or more detectors are further constructed and arranged to receive a plurality of pixel intensity values based, at least in part, upon detected emissions from a plurality of probe features disposed on the probe array at the best plane of focus; and
the system further comprises an image generator constructed and arranged to associate each of the pixel intensity values with one or more image pixel positions of a probe array based, at least in part, on comparison to a synthesis file.
2. The scanning system of claim 1 , wherein:
the reflected intensity data correspond to reflected light from one or more reflection spots.
3. The scanning system of claim 2 , wherein:
the best plane of focus is based, at least in part, upon associating the one or more reflection spots with one or more characteristics of a beam diameter.
4. A method, comprising the acts of:
directing an excitation beam at a probe array;
receiving reflected intensity data responsive to the excitation beam, wherein the intensity data is responsive, at least in part, to a focusing distance between an optical element and the probe array;
adjusting the focusing distance in a first direction with respect to the probe array; repeating the steps of receiving and adjusting for a number of iterations;
determining a best plane of focus based, at least in part, upon one or more characteristics of the reflected intensity data at the adjusted focusing distances;
dividing the reflected intensity values into one or more pixels to create pixel intensity values; and
generating an image of the probe array by associating each of the one or more pixel intensity values with one or more image pixel positions of the probe array based, at least in part, on comparison to a synthesis file.
5. The method of claim 4 , wherein:
the first direction is away from or toward the probe array.
6. The method of claim 4 , wherein:
the reflected intensity data are responsive to reflection of the excitation beam from one or more focus features.
7. The method of claim 6 , wherein:
the reflected intensity data correspond to one or more reflection spots.
8. The method of claim 7 , wherein:
the best plane of focus is based, at least in part, upon associating the one or more spots with one or more characteristics of a beam diameter.
9. The method of claim 7 , wherein:
the one or more focus features are positioned at one or more corners of the probe array.
10. A scanning system, comprising:
a probe array;
one or more optical elements constructed and arranged to direct an excitation beam at the probe array;
one or more detectors constructed and arranged to receive reflected intensity data responsive to the excitation beam, wherein the intensity data is determined, at least in part, by a focusing distance between an optical element and the probe array, and wherein the one or more detectors are further constructed and arranged to receive a plurality of pixel intensity values based, at least in part, upon detected emissions from a plurality of probe features disposed on the probe array at the best plane of focus;
an auto-focuser constructed and arranged to determine a best plane of focus based, at least in part, upon one or more characteristics of the reflected intensity data as received at two or more focusing distances, and
an image generator constructed and arranged to associate each of the pixel intensity values with one or more image pixel positions of a probe array based, at least in part, on comparison to a synthesis file.
11. The scanning system of claim 10 , wherein: the reflected intensity data are responsive to reflection of the excitation beam from one or more focus features.
12. The scanning system of claim 11 , wherein:
the one or more focus features are positioned at one or more corners of the probe array.
13. The scanning system of claim 11 , wherein:
the reflected intensity data correspond to one or more reflection spots.
14. The scanning system of claim 13 , wherein:
the best plane of focus is based, at least in part, upon associating the one or more spots with one or more characteristics of a beam diameter.