US 20050057756A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2005/0057756 A1 (43) Pub. Date: Fang-Yen et al. (54) SYSTEMS AND METHODS FOR PHASE MEASUREMENTS (52) U.S. c1. ............................................................ .. 356/497 (57) (75) Inventors: Christopher M. Fang-Yen, Somerville, MA (US); Gabriel Popescu, Brighton, MA (US); Changhuei Yang, Pasadena, CA (US); Adam Wax, Chapel Hill, NC (US); Ramachandra R. Dasari, Lexington, MA (US); Michael S. Feld, Newton, MA (US) Mar. 17, 2005 ABSTRACT Preferred embodiments of the present invention are directed to systems for phase measurement Which address the prob lem of phase noise using combinations of a number of strategies including, but not limited to, common-path inter ferometry, phase referencing, active stabilization and differ ential measurement. Embodiment are directed to optical devices for imaging small biological objects With light. These embodiments can be applied to the ?elds of, for Correspondence Address: WEINGARTEN, SCHURGIN, GAGNEBIN & LEBOVICI LLP example, cellular physiology and neuroscience. These pre ferred embodiments are based on principles of phase mea surements and imaging technologies. The scienti?c motiva tion for using phase measurements and imaging technologies is derived from, for example, cellular biology TEN POST OFFICE SQUARE BOSTON, MA 02109 (US) at the sub-micron level Which can include, Without limita (73) Assignee: Massachusetts Institute of Technology, Cambridge, MA neuronal transmission and implementation of the genetic (21) Appl. No.: 10/823,389 code. The structure and dynamics of sub-cellular constitu ents cannot be currently studied in their native state using the (22) Filed: Apr. 13, 2004 existing methods and technologies including, for example, Related US. Application Data (63) Continuation-in-part of application No. 10/024,455, ?led on Dec. 18, 2001. (60) tion, imaging origins of dysplasia, cellular communication, Provisional application No. 60/479,732, ?led on Jun. 19, 2003. x-ray and neutron scattering. In contrast, light based tech niques With nanometer resolution enable the cellular machinery to be studied in its native state. Thus, preferred embodiments of the present invention include systems based on principles of interferometry and/or phase measurements and are used to study cellular physiology. These systems include principles of 10W coherence interferometry (LCI) using optical interferometers to measure phase, or light scattering spectroscopy (LSS) Wherein interference Within the cellular components themselves is used, or in the alter native the principles of LCI and LSS can be combined to Publication Classi?cation (51) Int. Cl? ..................................................... .. G01B 9/02 result in systems of the present invention. 10 LI I: target sample 16 X1 D2 22 14 775 nm 34 32 <—L-—> <7— <——— BS 1550 nm M—> 12 composite low coherence 775 nm/CW 1550 nm beam Computer ,26 Patent Application Publication Mar. 17, 2005 Sheet 1 0f 74 US 2005/0057756 A1 mm X or PO>0EmE:Om\E“a?c /8 _ _ HoE mB._QE 823:.2908:Q< .wmoE:wn? $\$mt2. Fx NFvN @M55on w 8/“ \ mmN021| _ww\ ESQ O Patent Application Publication Mar. 17, 2005 Sheet 2 0f 74 50 US 2005/0057756 A1 52 interface light input > x FIG. 2 input 70 I‘ ht ‘g 1St interface 2nd interface 72 74 CW heterodyne signal 76 Z8 I. l I“ \ LC heterodyne signal / / 80 FIG. 3 A 84 Patent Application Publication Mar. 17, 2005 Sheet 3 0f 74 25 I I I US 2005/0057756 A1 I 20 15 b) v‘ 104 102 10 c 5 w “H H 106 —1OO O I “I a! l \1 -200 0 0 100 I 110 >1 I V ‘i. .' : * l 200 Min 300 400 500 . x (centered on ?rst interface) (pm) FIG. 4 600 Patent Application Publication Mar. 17, 2005 Sheet 4 0f 74 om? mo08 .3:8235.»60 H n u u EBPNQNSQWTN2VREKAVQ r5um2kaw“?*oMGmOEpLRw NE.we03 6?mg 893*NEEM + so BEénEs:F3IS.‘?7_fmwN© US 2005/0057756 A1 mm .OEm Patent Application Publication Mar. 17, 2005 Sheet 5 0f 74 US 2005/0057756 A1 124 Placing sample interfaces as the end /126 re?ectors of a signal interferometer arm l Scanning the reference mirror in the /12s reference interferometer arm l Combining the re?ections from the —> /130 signal and reference arms and separating them by wavelength l Detecting the heterodyne /132 oscillations in the intensities of the combined light l Extracting the phases of the /134 heterodyne signals l Evaluating a difference phase for /136 the entire scan l Repeating the scan with the /137 wavelength of the light being slightly detuned To Fl+G.6B FIG. 6A Patent Application Publication Mar. 17, 2005 Sheet 6 0f 74 US 2005/0057756 A1 From FIG. 6A i Superposing the two difference phases found from the two scans /138 1 Determining the locations of the /140 sample interfaces by the phase crossing points i Determining the optical separation between the interfaces by counting /142 the number of times the heterodyne signal associated with the CW light wraps over by 21: between the two crossing points i Re?ning the separation distance by /144 measuring the difference phase at the crossing points FIG. 6B Patent Application Publication Mar. 17, 2005 Sheet 7 0f 74 US 2005/0057756 A1 ow? v om? mmF\ 2E9Q5E3 893:62 80 >EmoOtm\:E.awc mm? H41. I¢t\ @2\ N9:v2 MR8“E: Ntwt\? N0 _ \ \A 1E 5598 82E:NRE: o5<wt/ _ oEwSQ\E Q Patent Application Publication Mar. 17, 2005 Sheet 8 0f 74 US 2005/0057756 A1 I, / 184 Placing sample interfaces as the end /186 re?ectors of a signal interferometer arm 1 Scanning the reference mirror in the /138 reference interferometer arm 1 Combining the re?ections from the /190 signal and reference arms and separating them by wavelength Further separating the LC wavelength-with ?lters ' Detecting the heterodyne /196 oscillations in the intensities of the \ l combined light Detecting the heterodyne i oscillations with three detectors Extracting the phases of the /198 heterodyne signals 1 Evaluating a difference phase for each LC signal with the CW signal /2oo i To FIG. 88 FIG. 8A Patent Application Publication Mar. 17, 2005 Sheet 9 0f 74 US 2005/0057756 A1 From FIG. 8A l Superposing the two difference /202 phases Determining the locations of the /204 sample interfaces by the phase crossing points Determining the optical separation between the interfaces by counting /206 the number of times the heterodyne signal associated with the CW light wraps over by 2n between the two crossing points i Re?ning the separation distance by measuring the difference phase at the crossing points FIG. 8B 208 / Patent Application Publication Mar. 17, 2005 Sheet 10 0f 74 US 2005/0057756 A1 wow @2w382w 4SE3w2ow C 8N DQB32E06O5mQ\)6B2@62862xL58 wm / 5N NmN ’TAHow868I.% > ES6mg. 522%i“302s mmw\.E b 202 .‘I’, v m8 NaH56o8w%SE \IY‘mmw! wow / EShQ\oEUQ mEw oE a.QE Patent Application Publication Mar. 17, 2005 Sheet 11 0f 74 / US 2005/0057756 A1 270 phase crossing system using low coherence 274 n 500 nm and 1000 nm U '’ CW light / 272 phase crossing system using low coherence n 900 nm and 1800 nm U CW light / 276 FIG. 10 278 ’ Patent Application Publication Mar. 17, 2005 Sheet 12 0f 74 300 / __ M~306 3?2 / 308 PM laser US 2005/0057756 A1 % 304% /31O : MM BS fast .... .§'.°.v.v...’5‘ 3 / C“' 312 L0 \320 317 display \/ /318 D / LO 322 feedback MX electronics 316/ FIG. 11 314 315 ( 1 computer Patent Application Publication Mar. 17, 2005 Sheet 13 0f 74 US 2005/0057756 A1 350 "(A / /352 laser 35 5 --_-----£.------.?. ........... - / 353 LC source 354 B3 375 364 AM LC display 373 ( feedback /L0 370 LO ’— 384 t electronics — comp“ er MX / 368 sample on coverslip W phase ~ ~ 386 FIG. 12 W 3% d Patent Application Publication Mar. 17, 2005 Sheet 14 0f 74 US 2005/0057756 A1 400 / Am,‘ W" r v 1 v -2nd M ?nv 11W! +2nd FIG. 13 AL Patent Application Publication Mar. 17, 2005 Sheet 16 0f 74 US 2005/0057756 A1 0 OEm50330‘ on.‘ L2BEcEm w \ \ now.\* won‘>m5w5 00 Ev 6=9.Eo Patent Application Publication Mar. 17, 2005 Sheet 17 0f 74 US 2005/0057756 A1 470 / LC2 \ALUV TV LCl *1 RM vnU UV VnvF|G.15A l MUM .l" 48° FIG. 15B AL Patent Application Publication Mar. 17, 2005 Sheet 18 0f 74 / US 2005/0057756 A1 500 d ‘L- Ef?l» sampl\e on coverslip T l C. f2 508 f1 502 "L211- _ __ LC2 \ 504 B LC2 I 526'] 91% D88 518 CCD or 530 '| LC1 516 V PD array Display D/524 525 L‘O 525 522 i Feedbapk electronlcs 525/MX FIG. 16 5,23 Computer Patent Application Publication Mar. 17, 2005 Sheet 19 0f 74 US 2005/0057756 A1 / 550 sample detected _ image a |e"$(f1) 5 \556 i, incidenti‘,_-' /-> i 54 \ " I: i E |en3(f2)\ plane g 558 g 560 ------ --<- --- ‘ beam ?“~~\‘__ 552 or oblectlve reference plane g \554 * . > ~ __:_ ______ __‘_h__\__ 5 f 562 f 1 mm I _5, 5 f 1 FIG. 17 I!‘ 2 \ I /> f >54 2 a‘
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