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(12) United States Patent (10) Patent No.: US 8,509,879 B2 Durkin Et Al

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(12) United States Patent (10) Patent No.: US 8,509,879 B2 Durkin Et Al US008509879B2 (12) United States Patent (10) Patent No.: US 8,509,879 B2 Durkin et al. (45) Date of Patent: Aug. 13, 2013 (54) APPARATUS AND METHOD FOR (56) References Cited WIDEFIELD FUNCTIONAL IMAGING (WIFI) USING INTEGRATED STRUCTURED U.S. PATENT DOCUMENTS LLUMINATION AND LASER SPECKLE 6,208.415 B1* 3/2001 De Boer et al. ............... 356,450 MAGING 2003/0002028 A1 1/2003 Rice et al. ....................... 356/39 2006, O155195 A1* 7, 2006 Maier et al. ........ ... 600,476 2006/0184043 A1* 8/2006 Tromberg et al. ..... ... 600,476 (75) Inventors: Anthony J. Durkin, Irvine, CA (US); 2006/0268241 A1* 11/2006 Watson et al. .................. 353/94 David Cuccia, Costa Mesa, CA (US); OTHER PUBLICATIONS Bruce J. Tromberg. Irvine, CA (US); Dunn, A. K. Simultaneous imaging of total cerebral hemoglobin Amaan Mazhar, Irvine, CA (US); concentration, oxygenation, and blood flow during functional acti Bernard Choi, Irvine, CA (US) vation, Optics Letters, vol. 28, 1 Jan 2003, pp. 28-30.* Briers, J.D., Laser Doppler, speckle, and related techniques for blood (73) Assignee: The Regents of the University of perfusion mapping and imaging. Physiological Measurement, 22 California, Oakland, CA (US) (2001), R35-R66.* MacKinnon, N., et al. Spectrally Programmable light engine for in (*) Notice: Subject to any disclaimer, the term of this vitro or in vivo molecular imaging and spectroscopy, Applied Optics, patent is extended or adjusted under 35 Apr. 10, 2005, vol. 44, No. 1 1. pp. 2033-2040.* U.S.C. 154(b) by 1294 days. Barlow, A.L., etal, Quantization of Widefield Fluorescence Imaging Using Structured Illumination and Image Analysis Software, Micros copy Research and Technique, 70: 76-84 (2007).* (21) Appl. No.: 12/016,918 Ventalon, C., et al., “Dynamic speckle illumination microscopy with Filed: Jan. 18, 2008 translated versus randomized speckle patterns. Opt. Express 14, (22) 71.98-7209 (2006).* (65) Prior Publication Data * cited by examiner US 2009/O118622A1 May 7, 2009 Primary Examiner — Unsu Jung Assistant Examiner — Daniel Huntley (74) Attorney, Agent, or Firm — Daniel L. Dawes; Marcus Related U.S. Application Data C. Dawes (60) Provisional application No. 60/985,872, filed on Nov. (57) ABSTRACT 6, 2007. An apparatus for wide-field functional imaging (WiFI) of (51) Int. C. tissue includes a spatially modulated reflectance/fluores A6 IB 6/00 (2006.01) cence imaging (SI) device capable of quantitative Subsurface (52) U.S. C. imaging across spatial scales, and a laser speckle imaging USPC ............ 600/473; 600/476; 600/479: 600/407 (LSI) device capable of quantitative subsurface imaging (58) Field of Classification Search across spatial scales using integrated with the (SI) device. The USPC ................. 600/473, 407,476, 479; 382/128; SI device and LSI device are capable of independently pro 250/459. 1; 35.9/368 viding quantitative measurement of tissue functional status. See application file for complete search history. 9 Claims, 20 Drawing Sheets 24 U.S. Patent Aug. 13, 2013 Sheet 1 of 20 US 8,509,879 B2 |-— • CN FIG. 1A U.S. Patent Aug. 13, 2013 Sheet 2 of 20 US 8,509,879 B2 DEMOD. & CAL. (EQS. 20, 23) DATA FIT (EQ. 10) FIG. 1B U.S. Patent Aug. 13, 2013 Sheet 3 of 20 US 8,509,879 B2 ORIGINAL SAMPLE FRINGE WRAPPED PATTERN GE_LOÍTHISNOOEYH LH5OIEH FIG. 1C U.S. Patent Aug. 13, 2013 Sheet 4 of 20 US 8,509,879 B2 ILLUMINATION: q = q (z) COS(kx+d) 1444444444444444444,A/A/A/A" X AA ?\. Afis 2 4-N-7.--N 7-N-7--Nys/A. Po(z) D C) e 1-NS- -- a-ara Tea as - A - a -- - - A -- an A. D H as-----> -----> -----As FLUENCE RATE: (p = p(z) COS(kx + d) FIG. 1D U.S. Patent Aug. 13, 2013 Sheet 5 of 20 US 8,509,879 B2 ABSORPTION VARIATION EXPERIMENT 0.002 mm guag 0.12 mm ob2 oboos oos o o2 o4 SPATIAL FREQUENCY, fx(mm) SCA TERING WARIATION EXPERIMENT 0.32 mm.< us''< 1.8 mm a = 0.0046 mm - P as 0.5 mm 0. 0.02 0.04 0.06 0.08 0.1 0.12 0.14 SPATIAL FREQUENCY, f(mm) FIG. 2 U.S. Patent Aug. 13, 2013 Sheet 7 of 20 US 8,509,879 B2 REFLECTANCE IMAGE + ROI (650 nm) ; AMPSAT 650 nm (I/nm) us' AMPSAT 650 nm (I/nm) 0.05 0.04 0.03 R. Sm. 0.02 MEAN=0.034382, SID=0.0048508 6000 ? 4000 D C O 2000 O O 0.02 0.03 0.04 O,O5 O,06 0.07 0.02 0.03 0.04 0.05 0.06 0.07 u (mm) us' (mm) FIG. 4B U.S. Patent Aug. 13, 2013 Sheet 8 of 20 US 8,509,879 B2 9.6 9.4 9.2 8.8 FIG. 5A 86 -5 0 5 O 15 20 25 30 TIME (min) 0.64 0.63 0.62 0.61 5 to 0.6 S. 0.59 FIG. 0.58 -5 O 5 10 15 20 25 30 TIME (min) U.S. Patent Aug. 13, 2013 Sheet 9 of 20 US 8,509,879 B2 AOHb (uM) AHHb (uM) ATHb (uM) Asto (%) 10 15 20 25 30 TIME (min) FIG. 5C U.S. Patent US 8,509,879 B2 ?I-(u?u)BWIL U.S. Patent Aug. 13, 2013 Sheet 11 of 20 US 8,509,879 B2 MAP, 650 nm 5mm FIG. 6A ABSORPTION VS. WAVELENGTH 0.055 O.05 0.045 0.04 0.035 0.03 FIG. 6B 0.025 0.02 0.015 0.0050.01 S 600 650 700 750 800 85O 900 950 1000 1050 WAVELENGTH (nm) REDUCED SCATTERING VS. WAVELENGTH 1.3 1.2 1.1 1. 0.9 FIG. 6C 0.8 0.7 0.6 0.5 600 650 700 750 800 85O 900 950 1000 1050 WAVELENGTH (nm) U.S. Patent Aug. 13, 2013 Sheet 12 of 20 US 8,509,879 B2 FIG. 7A 5 to 15 2025.303s 4045 m FLUORESCENCE (G) f = 0.21mm x10' FIG. 7B 5 O 15 20 25 30 35 40 45 n FLUORESCENCE (G) f = 0.45mm FIG. 7C 5 O 15 20 25 30 35 40 45 U.S. Patent Aug. 13, 2013 Sheet 13 of 20 US 8,509,879 B2 1. UM CY5.5 lar.01, his ~ 1. G) 660nm O. UM CY5.5 FIG. 7D INCREASING SPATIAL FREQUENCY O 2 3 4. 5 6 DISTANCE (mm) FIG. 7E. U.S. Patent Aug. 13, 2013 Sheet 14 of 20 US 8,509,879 B2 FLUORESCENCE IMAGE ABSORPTIONG 660nm f s 4 O.O2 3 5 8. S < S. 2 O. O. O. >- C O ne' 0.01 FIG. 6D "" ; ;QUADRANT ; ; U.S. Patent Aug. 13, 2013 Sheet 15 of 20 US 8,509,879 B2 VENULES TUMOR ARTERIOLES 30 mm/s is 8000 9, 7000 C 2 D O 2 N1 U a. f O 2 4 6 8 10 2 14 16 1820 O 2 3 4. 5 6 ACTUAL FLOW RATE mm/s) ACTUAL FLOW RATE mm/s) ? 500000 ?ts, 80000 400000 CY 2 2, 60000 as 300000 O 5 85 40000 3 200000 6 a. ein 20000 100000 T = 10 us i T = 100 us - l f O O O 50 100 150 200 250 300 O 50 100 150 200 250 300 ACTUAL FLOW RATE Imm/s) ACTUAL FLOW RATE Imm/s) FIG. 9 U.S. Patent Aug. 13, 2013 Sheet 16 of 20 US 8,509,879 B2 o Co o O o O Cd Co f C O N N o ser i. s U.S. Patent Aug. 13, 2013 Sheet 17 of 20 US 8,509,879 B2 ANALYSIS WORKSTATION FIG. 11A DUAL WAVELENGTHLED (785nm,820mm) IMAGING STAGE FIG. 11B L2 Ay LP 1 11 || Y IN NJ /1 || 1 U.S. Patent Aug. 13, 2013 Sheet 18 of 20 US 8,509,879 B2 FIG. 12A ANALYSIS WORKSTATION DUAL WAVELENGTH LASER (785nm,82Onm) CUSTOM LIGHT LIGHT GUIDE ENGINE (APOGEN) FILTER WHEEL BROADBAND SOURCE FILTER (TUNGSTEN HALOGEN) WHEEL IMAGING STAGE 8 ENCOSURE U.S. Patent Aug. 13, 2013 Sheet 19 of 20 US 8,509,879 B2 SNAPSHOT FIG. 13A ANALYSIS WORKSTATION DUAL WAVELENGTH LASER (785nm,820mm) BROADBAND SOURCE CUSTOM LIGHT ran-ras A re- IGHT GUIDE (TUNGSTEN HALOGEN) ENGINE (APOGEN) CTF CIS w M DUAL WAVELENGTH LED (785nm,820mm) ARTICULATING ARM FIG. 13B U.S. Patent Aug. 13, 2013 Sheet 20 of 20 US 8,509,879 B2 sassisssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssarsssssssssssssssssssssssssarrarararass s & US 8,509,879 B2 1. 2 APPARATUS AND METHOD FOR BRIEF SUMMARY OF THE INVENTION WIDEFIELD FUNCTIONAL IMAGING (WIFI) USING INTEGRATED STRUCTURED We disclose a wide-field functional imaging (WiFI) with LLUMINATION AND LASER SPECKLE the objective of developing an integrated imaging platform MAGING capable of quantitative Subsurface metabolic imaging across spatial scales. WiFI simultaneously measures tissue blood RELATED APPLICATIONS flow, biochemical composition (i.e. oxy- and deoxy-hemo globin, water and lipid content), and molecular fluorescence The present application is related to U.S. Provisional in turbid tissues. It possesses sufficient spatio-temporal reso Patent Application Ser. No. 60/985,872, filedon Nov. 6, 2007, 10 lution to study both fast (i.e., ms timescale) and localized (i.e., which is incorporated herein by reference and to which pri tens of um to mm) events at depths of several millimeters in ority is claimed pursuant to 35 USC 119. thick tissues. This platform enables quantitative insight into disease progression and therapeutic response in areas such as GOVERNMENT RIGHTS wound healing, neuroscience and cancer. 15 We disclose the design specifications and fabrication ele This invention was made with government Support under ments necessary to construct a series of instruments based on grant RR001 192 awarded by the National Institutes of the integration of two wide-field imaging modalities; Spa Health.
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