System and method for fluorescence detection
A system and method for determining fluorescence decay in a biological sample is provided. The method includes acquiring optical signal data from at least a part of a biological sample undergoing fluorescence, assembling the optical signal data into a set of spatial time-series data, and converting the set of spatial time-series data into a set of spatial frequency time-series data. The method also includes applying a spatial filter to the set of spatial frequency time-series data to yield a set of filtered frequency-series data, the spatial filter configured to separate, from the set of frequency-series data, fluorescence signals consistent with a non-diffuse component and converting the set of filtered frequency-series data into a set of filtered time-series data. The method further includes determining, using the filtered time-series data, a fluorescence signature consistent with fluorescence decay of at least one fluorophore, and generating a report indicative of the fluorescence signature of the biological sample.
1. A method for determining fluorescence decay in a biological sample, the method comprising:
acquiring optical signal data from at least one part of a biological sample undergoing fluorescence;
assembling the optical signal data into a set of spatial time-series data;
converting the set of spatial time-series data into a set of spatial frequency time-series data;
applying a spatial filter to the set of spatial frequency time-series data to yield a set of filtered spatial frequency time-series data, the spatial filter configured to separate, from the set of spatial frequency time-series data, fluorescence signals consistent with a non-diffuse component;
converting the set of filtered spatial frequency time-series data into a set of filtered spatial time-series data;
determining, using the filtered spatial time-series data, a fluorescence signature consistent with fluorescence decay of at least one fluorophore in the biological sample; and
generating a report indicative of the fluorescence signature corresponding to the biological sample.
2. The method of claim 1 , the method further comprises creating an excitation in the at least one part of biological sample using a point-like light source.
3. The method of claim 1 , the method further comprises creating an excitation in the at least one part of biological sample using a pulsed light source.
4. The method of claim 1 , the method further comprises creating an excitation in the at least one part of biological sample using a spatially modulated light source.
5. The method of claim 1 , wherein assembling the set of spatial time-series data further comprises performing a threshold processing using a maximum intensity of signals associated with the optical signal data.
6. The method of claim 1 , wherein converting the set of spatial time-series data includes performing a Fourier transform for a plurality of time points.
7. The method of claim 1 , wherein the spatial filter is dependent upon a fluorescence decay profile determined by optical properties of the biological sample.
8. The method of claim 1 , wherein the spatial filter is a band-pass filter that minimizes fluorescence signals described by spatial frequencies consistent with a diffuse component.
9. The method of claim 1 , wherein converting the set of filtered spatial frequency time-series data includes performing an inverse Fourier transform for a plurality of time points.
10. The method of claim 1 , the method further comprises performing a curve fitting process, using the filtered spatial time-series data, to determine the fluorescence decay of the at least one fluorophore.
11. The method of claim 1 , wherein the report includes at least one lifetime map obtained using the fluorescence decay of the at least one fluorophore.
12. The method of claim 1 , wherein the fluorescence decay of the at least one fluorophore is shorter than an intrinsic temporal response of a diffuse medium including the at least one part of the biological sample undergoing fluorescence.
13. A system for determining fluorescence decay in a biological sample, the system comprising:
an input designed to receive optical signal data from at least one part of a biological sample undergoing fluorescence;
at least one processor configured to:
assemble the optical signal data into a set of spatial time-series data;
convert the set of time-series data into a set of spatial frequency times-series data;
apply a spatial filter to the set of spatial frequency time-series data to yield a set of filtered spatial frequency time-series data, the spatial filter configured to
separate, from the set of spatial frequency time-series data, fluorescence signals consistent with a non-diffuse component;
convert the set of filtered spatial frequency time-series data into a set of filtered spatial time-series data;
determine, using the filtered spatial time-series data, a fluorescence signature consistent with fluorescence decay of at least one fluorophore in the biological sample; and
generate a report indicative of the fluorescence signature corresponding to the biological sample.
14. The system of claim 13 , the system further comprising an excitation source for creating an excitation in the at least one part of biological sample.
15. The system of claim 14 , the excitation source configured to create an excitation defined by at least one of a point-like light excitation, a pulsed light excitation, and a spatially modulated excitation.
16. The system of claim 13 , wherein the at least one processor is further configured to perform a threshold processing using a maximum intensity of signals associated with the optical signal data.
17. The system of claim 13 , wherein the at least one processor is further configured to perform a Fourier transform, using the set of spatial time-series data, for a plurality of time points.
18. The system of claim 13 , wherein the spatial filter is dependent upon a fluorescence decay profile determined by optical properties of the biological sample.
19. The system of claim 13 , wherein the spatial filter is a band-pass filter that minimizes fluorescence signals described by spatial frequencies consistent with a diffuse component.
20. The system of claim 13 , wherein the at least one processor is further configured to perform an inverse Fourier transform, using the set of filtered spatial frequency time-series data, for a plurality of time points.
21. The system of claim 13 , wherein the at least one processor is further configured to perform a curve fitting process, using the filtered spatial time-series data, to determine the fluorescence decay of the at least one fluorophore.
22. The system of claim 13 , wherein the report includes at least one lifetime map obtained using the fluorescence decay of the at least one fluorophore.
23. The system of claim 13 , where the fluorescence decay of the at least one fluorophore is shorter than an intrinsic temporal response of a diffuse medium including the at least one part of the biological sample undergoing fluorescence.
24. A method for determining fluorescence decay in a biological sample, the method comprising:
assembling a set of spatial time-series data using optical signals acquired from a biological sample having a plurality of fluorophores;
converting the set of spatial time-series data into a set of spatial frequency time-series data;
applying a filter to the set of spatial frequency time-series data to yield a set of filtered spatial frequency time-series data, the filter configured to separate, from the set of spatial frequency time-series data, fluorescence signals consistent with a non-diffuse component;
converting the set of filtered spatial frequency time-series data into a set of filtered spatial time-series data;
determining, using the filtered spatial time-series data, a fluorescence signature consistent with fluorescence decay of at least one fluorophore in the plurality of fluorophores; and
generating a report indicative of the fluorescence signature corresponding to the biological sample.
25. The method of claim 24 , the method further comprises creating an excitation in the biological sample using a point-like light source.
26. The method of claim 24 , the method further comprises creating an excitation in the biological sample using a pulsed light source.
27. The method of claim 24 , the method further comprises creating an excitation in the biological sample using a spatially modulated light source.
28. The method of claim 24 , wherein assembling the set of spatial time-series data further comprises performing a threshold processing using a maximum intensity of signals associated with the optical signals acquired from the biological sample.
29. The method of claim 24 , wherein converting the set of spatial time-series data includes performing a Fourier transform for a plurality of time points.
30. The method of claim 24 , wherein the filter is dependent upon a fluorescence decay profile determined by optical properties of the biological sample.
31. The method of claim 24 , wherein the filter is a band-pass filter that minimizes fluorescence signals described by spatial frequencies consistent with a diffuse component.
32. The method of claim 24 , wherein converting the set of filtered spatial frequency time-series data includes performing an inverse Fourier transform for a plurality of time points.
33. The method of claim 24 , the method further comprises performing a curve fitting process, using the filtered spatial time-series data, to determine the fluorescence decay of the at least one fluorophore.
34. The method of claim 24 , wherein the report includes at least one lifetime map obtained using the fluorescence decay of the at least one fluorophore.
35. The method of claim 24 , where the fluorescence decay of the at least one fluorophore is shorter than an intrinsic temporal response of a diffuse medium including the biological sample.