Methods and systems of optical imaging for target detection in a scattering medium
View Patent ↗A method for enhancing contrast in fluorescence imaging is provided. The method comprises providing a patterned illumination source for illuminating one or more regions corresponding to a scan step, scanning at least a portion of a surface of a subject using a plurality of scan steps, acquiring image frames corresponding to two or more scan steps, deducting a background fluorescence from the image frames corresponding to the two or more scan steps to form one or more processed image frames, and reconstructing an image using one or more of the processed image frames.
1. A method for enhancing contrast in fluorescence imaging, comprising:
providing a patterned illumination source for illuminating one or more regions corresponding to a scan step;
scanning at least a portion of a surface of a subject using a plurality of scan steps;
collecting an emitted light from an illuminated region as well as from an area of interest away from the illuminated region;
acquiring image frames corresponding to two or more scan steps;
deducting a background fluorescence from the image frames corresponding to the two or more scan steps to form one or more processed image frames, wherein the background fluorescence is based on an excitation spread function; and
reconstructing an image using one or more of the processed image frames.
2. The method of claim 1 , wherein the step of deducting comprises:
collecting an emitted spectrum from a first illumination region;
collecting an excitation spread function for the first illumination region; and
deducting the excitation spread function from the emitted spectrum.
3. The method of claim 1 , wherein an illumination the illuminated region is illuminated by a patterned illumination, and wherein the patterned illumination comprises a point, an array of points, a line, an array of lines, a grid, a non-solid pattern extending along the surface, or combinations thereof.
4. The method of claim 1 , wherein at least one illumination region comprises a target, and wherein one or more non-target illumination regions are used to assess the background fluorescence.
5. The method of claim 1 , wherein the background fluorescence is expressed as an empirical equation, and wherein the empirical equation comprises a Lorentzian fit, a Gaussian fit, or a polynomial fit.
6. The method of claim 1 , further comprising administering a contrast agent into a scattering medium.
7. The method of claim 1 , wherein reconstructing the image comprises summing together the processed image frames to generate a composite image.
8. The method of claim 1 , further comprising using a noise reduction algorithm prior to or as a part of image reconstruction.
9. The method of claim 1 , wherein acquiring the image frames corresponding to the two or more scan steps comprises moving the illumination source relative to the subject while acquiring the image frames.
10. The method of claim 1 , wherein acquiring the image frames corresponding to the two or more scan steps comprises moving the subject relative to the illumination source.
11. The method of claim 1 , wherein acquiring the image frames comprises descanning the image frames corresponding to the two or more scan steps.
12. The method of claim 1 , wherein reconstructing the image comprises summing up the one or more processed image frames.
13. A fluorescence imaging system for generating an enhanced image of a target in a subject, comprising:
a source configured to illuminate at least a portion of a surface of the subject by a patterned illumination at one or more scan steps;
an imaging detector configured to acquire image frames at one or more scan steps, wherein the imaging detector is configured to collect emitted light from an illuminated region as well as from an area of interest away from the illuminated region; and
an imaging processor configured to process the image frames to deduct a background fluorescence from the image frames and reconstruct an image of the target using the processed image frames, wherein the background fluorescence is based on an excitation spread function.
14. The system of claim 13 , wherein the imaging detector is a one dimensional detector.
15. The system of claim 13 , wherein the imaging detector is selected from a group consisting of a charged coupled device, an intensified charged coupled device, a time-gated charged coupled device, a gain-modulated charged coupled device, a complementary metal oxide semiconductor device, an electron bombardment charge coupled device, and an image intensifier tube.
16. The system of claim 13 , wherein the source comprises a substantially monochromatic light source.
17. The system of claim 13 , wherein the source comprises a continuous wave light source, a pulsed laser, a frequency modulated light source, an intensity modulated light source, a phase varying light source, or combinations thereof.
18. The system of claim 13 , further comprising an illumination deflection device for directing the patterned illumination onto the surface of the subject.
19. The system of claim 18 , wherein the illumination deflection device is in operative association with the detector to enable a descanned arrangement.
20. The system of claim 13 , further comprising collection optics for directing the emitted light to the detector.