Method and detection system for detection of aflatoxin in corn with fluorescence spectra
A system and method for fluorescence spectral imaging of target material to detect the presence of a contaminant (such as aflatoxin in corn) is provided. An ultraviolet light source is coupled with a light-excluding compartment. The fluorescence from the UV excited target passes through a filter (liquid crystal tunable, acoustic-optic tunable, a filter wheel, or other wavelength splitting device) and a lens, to a spectral imaging camera. Fluorescence spectral image data from the camera are analyzed by a computer and presented in human-readable form. Aflatoxin detection in contaminated corn kernels is based on peak fluorescence and peak fluorescence shift in the spectral range from 451 nm to 500 nm. Aflatoxin contamination level within the target material is quantified based on peak fluorescence and peak fluorescence shift and computed corn kernel pixel statistics.
1. A method for aflatoxin detection using fluorescence spectral images, comprising the steps of: introducing target material before an ultraviolet light source; exciting the target material with ultraviolet light from the ultraviolet light source; passing fluorescence generated by the ultraviolet light excitation of the target material through a focusing device and a wavelength splitting optical device; detecting wavelength split fluorescence from the focusing device and the wavelength splitting optical device with a spectral data acquisition device which generates fluorescence spectral image data from the wavelength split fluorescence; transferring the fluorescence spectral image data from the spectral data acquisition device to a computer analysis system; processing the fluorescence spectral image data with the computer analysis system, wherein the processing includes pre-processing the fluorescence spectral image data with the computer system, identifying pixels associated with the target material in the fluorescence spectral images, computing statistics of spectral features for each identified target material pixel, characterizing fluorescence spectral features of each identified target material pixel, identifying peak fluorescence and peak fluorescence shift features within the characterized fluorescence spectral features, identifying aflatoxin contaminated target material pixels based on identified peak fluorescence and identified peak fluorescence shift features, and quantifying aflatoxin contamination levels within the imaged target material based on identified peak fluorescence and identified peak fluorescence shift features and computed target material pixel statistics; and presenting the analyzed fluorescence spectral imaging data regarding target material aflatoxin contamination detection and quantification results in human-readable form.
2. The method of claim 1 , wherein
the target material is corn, and
the ultraviolet light wavelength is centered at approximately 365 nm.
3. The method of claim 2 , wherein
the ultraviolet light source includes an ultraviolet lamp or ultraviolet light-emitting-diode array (UV-LED) and a band pass fluorescence excitation filter, and
the ultraviolet lamp or ultraviolet light-emitting-diode array and the band pass fluorescence excitation filter wavelengths are both centered at approximately 365 nm.
4. The method of claim 1 , wherein the wavelength splitting optical device includes
a wavelength splitting optical instrument, and
a front-mounted long pass emission filter which blocks transmission of ultraviolet light with wavelength less than 400 nm.
5. The method of claim 4 , wherein
the wavelength splitting optical instrument is one of a spectrograph, a set of separate filters having pre-defined wavelengths, or a tunable filter such as a liquid crystal tunable filter, an acoustic-optic tunable filter, or a filter wheel with a set of filters having pre-defined wavelengths.
6. The method of claim 1 , wherein
the spectral data acquisition device includes an imaging camera or photodiode detector array, and
the imaging camera or photodiode detector array have an effective spectral response from 400 to 1000 nm.
7. The method of claim 1 , wherein the pre-processing includes
dark current removal,
image calibration to convert from radiance values received at the spectral data acquisition device to fluorescence values of the target material,
spectral noise smoothing, and
noisy band removal.
8. The method of claim 1 , wherein the spectral features are fluorescence spectrum values for each identified target material pixel.
9. The method of claim 2 , wherein the characterizing fluorescence spectral features of each aflatoxin contaminated corn pixel is in the spectral range from 451 nm to 500 nm.
10. The method of claim 2 , wherein the peak fluorescence and peak fluorescence shift features is in the spectral range from 451 nm to 500 nm.
11. The method of claim 2 , wherein
aflatoxin contaminated corn kernel pixels are identified based on peak fluorescence and peak fluorescence shift features, where healthy, uncontaminated corn has fluorescence features shifted toward shorter wavelength and aflatoxin contaminated corn has fluorescence features shifted toward longer wavelength in the spectral range from 451 nm to 500 nm.
12. The method of claim 1 , wherein
the quantification of aflatoxin contamination is further based on identified target material pixel statistics.
13. The method of claim 2 , wherein the presenting step includes
presenting aflatoxin contaminated corn detection and quantification results including a corn sample grading report based on published aflatoxin regulations using different ppb (parts per billion) levels for differentiation, and aflatoxin contamination level for each corn sample.
14. A system for the detection of aflatoxin using fluorescence spectral imaging, comprising: at least one ultraviolet light source; at least one band pass excitation filter; a compartment for containing ultraviolet light generated by the at least one ultraviolet light source and excluding outside light, the at least one ultraviolet light source being located within the compartment such that light emitted from the at least one ultraviolet light source passes through the at least one band pass excitation filter and is incident upon target material which is introduced into the compartment; a spectral data acquisition device which detects fluorescence generated by excitation of the target material by the ultraviolet light and generates fluorescence spectral image data; at least one emission filter to block ultraviolet light with wavelength less than 400 nm from entering the spectral data acquisition device; at least one wavelength splitting optical instrument through which fluorescence generated by excitation of the target material by the ultraviolet light passes prior to entering the spectral data acquisition device; and a computer analysis system which receives and analyzes the fluorescence spectral image data generated by the spectral data acquisition device and presents at least a portion of the aflatoxin detection and quantification results of the analyzed fluorescence spectral image data in human-readable form, wherein the fluorescence spectral image data includes identified peak fluorescence and identified peak fluorescence shift features.
15. The system for the detection of aflatoxin using fluorescence spectral imaging of claim 14 , wherein
the target material is corn;
the at least one ultraviolet light source wavelength is centered at approximately 365 nm;
the at least one band pass excitation filter is set at approximately 365 nm; and
the fluorescence spectral image data is in the spectral range from 451 nm to 500 nm.
16. The system for the detection of aflatoxin using fluorescence spectral imaging of claim 14 , wherein the compartment includes
an aperture arranged to receive the fluorescence generated from the target material, and
a platform upon which the target material resides during excitation by the at least one ultraviolet light source.
17. The system for the detection of aflatoxin using fluorescence spectral imaging of claim 14 , wherein
the platform is a conveyor which conveys the target material into and out of the compartment and into the field of view of the spectral data acquisition device.
18. The system for the detection of aflatoxin using fluorescence spectral imaging of claim 14 , wherein
the platform is a tray that holds the target material in the field of view of the spectral data acquisition device.
19. The system for the detection of aflatoxin using fluorescence spectral imaging of claim 14 , wherein
the spectral data acquisition device includes an imaging camera or photo-diode detector array.
20. The system for the detection of aflatoxin using fluorescence spectral imaging of claim 14 , wherein
the at least one wavelength splitting optical instrument is one of a spectrograph, a set of separate filters having pre-defined wavelengths, or a tunable filter such as a liquid crystal tunable filter, an acoustic-optic tunable filter, or a filter wheel with a set of filters having pre-defined wavelengths.
21. The system for the detection of aflatoxin using fluorescence spectral imaging of claim 15 , wherein
the at least one ultraviolet light source includes an ultraviolet lamp or ultraviolet light-emitting-diode array (UV-LED) and a band pass fluorescence excitation filter, and
the ultraviolet lamp or ultraviolet light-emitting-diode array and the band pass fluorescence excitation filter wavelengths are both centered at approximately 365 nm.
22. A portable system for the detection of aflatoxin using fluorescence spectral imaging, comprising: a portable power source; an ultraviolet light source; at least one band pass excitation filter; a compartment for containing ultraviolet light generated by the ultraviolet light source and excluding outside light, the ultraviolet light source being located within the compartment such that light emitted from the ultraviolet light source passes through the band pass excitation filter and is incident upon target material which is introduced into the compartment; a spectral data acquisition device includes a photo-diode detector array which detects fluorescence generated by excitation of the target material by the ultraviolet light and generates fluorescence spectral image data; an inserter for inserting the target material into and out the compartment and into the field of view of the spectral data acquisition device; at least one emission filter to block ultraviolet light with wavelength less than 400 nm from entering the spectral data acquisition device; a wavelength splitting optical instrument includes a set of separate filters having pre-defined wavelengths through which fluorescence generated by excitation of the target material by the ultraviolet light passes prior to entering the spectral data acquisition device; and a computer analysis system which receives and analyzes the fluorescence spectral image data generated by the spectral data acquisition device and presents at least a portion of the aflatoxin detection and quantification results of the analyzed fluorescence spectral image data in human-readable form, wherein the ultraviolet light source, the band pass excitation filter, the spectral data acquisition device, the at least one emission filter, and the wavelength splitting optical instrument are located within the compartment, and the fluorescence spectral image data includes identified peak fluorescence and identified peak fluorescence shift features.
23. The portable system for the detection of aflatoxin using fluorescence spectral imaging of claim 22 , wherein
the target material is corn;
the ultraviolet light source wavelength is centered at approximately 365 nm;
the at least one band pass excitation filter is set at approximately 365 nm; and
the fluorescence spectral image data is in the spectral range from 451 nm to 500 nm.
24. The portable system for the detection of aflatoxin using fluorescence spectral imaging claim 22 , wherein the inserter includes
an inserter handle arranged to support the target material; and
an inserter platform arranged to act as a seal to contain the ultraviolet light and exclude outside light from entering the compartment when the inserter is fully inserted into the compartment.
25. The portable system for the detection of aflatoxin using fluorescence spectral imaging of claim 22 , wherein
the inserter is a tray that holds the target material in the field of view of the spectral data acquisition system.
26. The portable system for the detection of aflatoxin using fluorescence spectral imaging of claim 22 , wherein
the spectral data acquisition device includes a photodiode detector array or a micro-spectrometer.
27. The portable system for the detection of aflatoxin using fluorescence spectral imaging of claim 23 , wherein
the ultraviolet light source includes an ultraviolet light-emitting-diode array (UV-LED) and a band pass fluorescence excitation filter, and
the ultraviolet light-emitting-diode array and the band pass fluorescence excitation filter are both centered at approximately 365 nm.