Absolute intensity determination for a fluorescent light source in low level light imaging systems
First Claim
1. A method for generating absolute fluorescent light intensity data for a fluorescent light source being imaged at low intensity light levels, the method comprising:
- determining a compensation factor for converting data output by a camera into absolute fluorescent intensity light data, the compensation factor corresponding to a set of imaging conditions;
storing the compensation factor;
calling the compensation factor based on the set of imaging conditions when the set of imaging conditions are used in imaging the fluorescent light source; and
modifying image data, output by the camera, by the compensation factor to obtain the absolute fluorescent light intensity data for the fluorescent light source, wherein the fluorescent light source emits light with an intensity in the range of about 103 to about 1010 photons/second/centimeter squared/steradian, and wherein the fluorescent light source includes fluorescing molecules.
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Abstract
The invention describes systems and methods to obtain and present imaging data in absolute units. The systems and methods convert relative image data produced by a camera to absolute light intensity data using a compensation factor. The compensation factor accommodates for hardware and specific imaging conditions in the imaging system that variably affect camera output. The present invention determines the compensation factor based on assessing the output of the camera against a known light source for a specific set of imaging conditions in the imaging system. The compensation factor is then stored in memory corresponding to the specific set of imaging conditions. Upon subsequent imaging with the set of imaging conditions, the corresponding compensation factor is called from memory and applied to the camera output. A compensation factor may be determined and stored for each hardware state and imaging condition available to the imaging system.
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Citations
19 Claims
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1. A method for generating absolute fluorescent light intensity data for a fluorescent light source being imaged at low intensity light levels, the method comprising:
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determining a compensation factor for converting data output by a camera into absolute fluorescent intensity light data, the compensation factor corresponding to a set of imaging conditions; storing the compensation factor; calling the compensation factor based on the set of imaging conditions when the set of imaging conditions are used in imaging the fluorescent light source; and modifying image data, output by the camera, by the compensation factor to obtain the absolute fluorescent light intensity data for the fluorescent light source, wherein the fluorescent light source emits light with an intensity in the range of about 103 to about 1010 photons/second/centimeter squared/steradian, and wherein the fluorescent light source includes fluorescing molecules. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method for imaging a light source at low intensity light levels in an imaging system including a camera and an imaging box, the method comprising:
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placing a portable light calibration device in the imaging box, said portable light calibration device having been calibrated against a known absolute intensity light source; setting one or more image capture conditions in the imaging system; determining a compensation factor that accommodates for differences between light emitted from the portable light calibration device and data output by the camera based on said one or more image capture conditions; removing said portable light calibration device from the imaging box; placing a light source which is to be imaged in the imaging box; capturing image data from the light source using the camera; and converting image data produced by the camera to absolute light intensity data for the light source using said compensation factor. - View Dependent Claims (16, 17, 18)
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19. A computer readable medium including instructions for generating absolute light intensity data for a fluorescent light source being imaged at low intensity light levels in an imaging box, the instructions comprising:
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instructions for determining a compensation factor for convening relative intensity data output by a camera into absolute light data, the compensation factor corresponding to differences between light emitted from a portable light calibration device and data output by the camera for a set of imaging conditions within the imaging box; instructions for storing the compensation factor; instructions for calling the compensation factor based on the set of imaging conditions when the set of imaging conditions are used in imaging the light source within the imaging box; and instructions for modifying image data, output by the camera, by the compensation factor to obtain the absolute light intensity data for the light source.
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Specification