Light modulated microarray reader and methods relating thereto
AssigneeDigital Optical Imaging Corporation
InventorsCalum E. MacAulay, Pierre M. Lane
Life of this patent
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Abstract
Microarray readers and methods that compensate for target spots that are too dim or too bright for the microarray reader to accurately measure. The readers adjust the amount of light directed at or received from specific non-acceptable target spots, such that dim spots receive more excitation light and overly bright spots receive less. This increases or decreases, respectively, their measured brightness, which in turn effectively increases the range over which a microarray reader can accurately measure the spots, and can also improve the signal-to-noise ratio and other aspects of the measurements.
Claims64
An automated method of reading a microarray comprising,
- a) providing an initial representation of a microarray comprising a plurality of target spots illuminated by illumination light having a designated intensity;
- b) determining from the initial representation whether at least one of the target spots has an emanating light intensity that is not between selected upper and lower threshold values, and designating such target spot a non-acceptable target spot;
- and,
- c) modulating the designated intensity of the illumination light via an automated upstream selective light modulator located in an illumination light path substantially at a conjugate image plane of the sample to provide a modulated illumination light and an adjusted target spot that emanates an adjusted light intensity between the selected upper and lower threshold values.
An automated method of reading a microarray comprising,
- a) providing an initial representation of a microarray comprising a plurality of target spots illuminated by illumination light having a designated intensity;
- b) determining from the initial representation whether at least one of the target spots has an emanating light intensity that is not between selected upper and lower threshold values, and designating at least one of such target spots a non-acceptable target spot;
- and,
- c) modulating the emanating light intensity via an automated downstream selective light modulator located in a detection light path substantially at a conjugate image plane of the sample to provide a modulated detection light comprising an adjusted emanating light intensity that is between the selected upper and lower threshold values.
A microarray reader comprising an automated upstream selective light modulator located upstream of a microarray in an illumination light path substantially at a conjugate image plane of the sample, and a light detector disposed downstream from the microarray in a detection light path substantially at a conjugate image plane of the sample, wherein the selective light modulator and the light detector are operably connected to at least one controller containing computer-implemented programming that controls transmissive characteristics of the upstream selective light modulator and that compiles an amount of modulated illumination light when the upstream selective light modulator is modulated and an adjusted light intensity emanating from a target spot on a microarray receiving the modulated illumination light, and wherein the controller selectively varies the transmissive characteristics of the selective light modulator to vary the modulated illumination light impinging on at least one non-acceptable target spot of the microarray such that light emanating from the at least one non-acceptable target spot is between selected threshold levels.
A microarray reader comprising an automated downstream selective light modulator located downstream of a microarray in a detection light path substantially at a conjugate image plane of the sample, and a light detector disposed in the detection light path substantially at a conjugate image plane of the sample and downstream from the downstream selective light modulator and the microarray, wherein the downstream selective light modulator and the light detector are operably connected to at least one controller containing computer-implemented programming that controls transmissive characteristics of the downstream selective light modulator and that compiles an amount of modulated detection light when the downstream selective light modulator is modulated and an adjusted light intensity received by the detector, and wherein the controller selectively varies the transmissive characteristics of the downstream selective light modulator to vary the modulated detection light emanating from at least one non-acceptable target spot of the microarray such that light received at the detector from the at least one non-acceptable target spot is between selected threshold levels.
An automated method of reading a microarray comprising,
- a) providing an initial representation of a microarray comprising a plurality of target spots illuminated by illumination light having a designated intensity;
- b) determining from the initial representation whether at least one of the target spots has an emanating light intensity that is not between selected upper and lower threshold values, and designating at least one of such target spots as a non-acceptable target spot;
- c) selectively illuminating the non-acceptable target spot via selectively transmitting light to the microarray using a first automated upstream selective light modulator located in an illumination light path substantially at a conjugate image plane of the sample;
- and,
- d) modulating the designated intensity of the illumination light via a second automated upstream selective light modulator located in the illumination light path substantially at a conjugate image plane of an aperture diaphragm of the objective lens, to provide a modulated illumination light and an adjusted target spot that emanates an adjusted light intensity between the selected upper and lower threshold values.
An automated method of reading a microarray comprising,
- a) providing an initial representation of a microarray comprising a plurality of target spots illuminated by illumination light having a designated intensity;
- b) determining from the initial representation whether at least one of the target spots has an emanating light intensity that is not between selected upper and lower threshold values, and designating at least one of such target spots as a non-acceptable target spot;
- c) selectively detecting light from the non-acceptable target spot via selectively transmitting light from the microarray using a first automated downstream selective light modulator located in a detection light path substantially at a conjugate image plane of the sample;
- and,
- d) modulating the emanating light intensity via a second automated downstream selective light modulator located in a detection light path substantially at a conjugate image plane of an aperture diaphragm of the objective lens, to provide a modulated detection light comprising an adjusted emanating light intensity between the selected upper and lower threshold values.
A microarray reader comprising a first automated upstream selective light modulator located upstream of a microarray in an illumination light path substantially at a conjugate image plane of the sample, a second automated upstream selective light modulator located upstream of the microarray in the illumination light path substantially at a conjugate image plane of an aperture diaphragm of the objective lens, and a light detector disposed downstream from the microarray in a detection light path substantially at a conjugate image plane of the sample, wherein the first and second selective light modulators and the light detector are operably connected to at least one controller containing computer-implemented programming that controls transmissive characteristics of the first and second upstream selective light modulators and that compiles an amount of modulated illumination light when the second upstream selective light modulator is modulated and an adjusted light intensity emanating from a target spot on a microarray receiving the modulated illumination light, and wherein the controller selectively varies the transmissive characteristics of the second selective light modulator to vary the modulated illumination light impinging on at least one non-acceptable target spot of the microarray such that light emanating from the at least one non-acceptable target spot is between selected threshold levels.
A microarray reader comprising a first automated downstream selective light modulator located downstream of a microarray in a detection light path substantially at a conjugate image plane of the sample, a second automated downstream selective light modulator located downstream of the microarray in the illumination light path substantially at a conjugate image plane of an aperture diaphragm of the objective lens, and a light detector disposed in a detection light path substantially at a conjugate image plane of the sample and downstream from the first and second downstream selective light modulators and the microarray, wherein the first and second selective light modulators and the light detector are operably connected to at least one controller containing computer-implemented programming that controls transmissive characteristics of the first and second downstream selective light modulator and that compiles an amount of modulated detection light when the second downstream selective light modulator is modulated and an adjusted emanating light intensity received by the detector, and wherein the controller selectively varies the transmissive characteristics of the second downstream selective light modulator to vary the modulated detection light emanating from at least one non-acceptable target spot of the microarray such that light received at the detector from the at least one non-acceptable target spot is between selected threshold levels.
Dependencies are inferred from the claim text.