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(12) Patent Application Publication (10) Pub. No.: US 2010/0105035 A1 Hashsham Et Al US 20100105035A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2010/0105035 A1 Hashsham et al. (43) Pub. Date: Apr. 29, 2010 (54) ELECTROLUMINESCENT-BASED Related U.S. Application Data FLUORESCIENCE DETECTION DEVICE (60) Provisional application No. 60/860,702, filed on Nov. 22, 2006. (76) Inventors: Syed Anwar Hashsham, Okemos, MI (US); James M. Tiedje, Publication Classification Lansing, MI (US); Erdogan (51) Int. Cl. Gulari, Ann Arbor, MI (US); Dieter CI2O I/68 (2006.01) Tourlousse, East Lansing, MI (US); GOI. I./S8 (2006.01) Robert Stedtfeld, Lansing, MI C4DB 60/2 (2006.01) (US); Farhan Ahmad, East CI2M I/34 (2006.01) Lansing, MI (US); Gregoire CI2M I/240 (2006.01) Seyrig, Lansing, MI (US); Onnop CI2O I/04 (2006.01) Srivannavit, Ann Arbor, MI (US) CI2P 19/34 (2006.01) (52) U.S. Cl. ......... 435/6; 250/458.1506/39; 435/288.7; Correspondence Address: 250/459. 1; 435/34: 435/91.2 MEDLEN & CARROLL, LLP (57) ABSTRACT 101 HOWARD STREET, SUITE 350 The present invention provides compositions providing and SAN FRANCISCO, CA 94105 (US) methods using fluorescence detection device, comprising an electroluminescent light (EL) source, for measuring fluores (21) Appl. No.: 12/312,686 cence in biological samples. In particularly preferred embodiments, the present invention provides an economical, battery powered and Hand-held device for detecting fluores (22) PCT Filed: Nov. 21, 2007 cent light emitted from reporter molecules incorporated into DNA, RNA, proteins or other biological samples, such as a (86). PCT No.: PCT/US07/24290 fluorescence emitting biological sample on a microarray chip. Further, a real-time hand-held PCR Analyzer device S371 (c)(1), comprising an EL light source for measuring fluorescence (2), (4) Date: Dec. 10, 2009 emissions from amplified DNA is provided. B 10 19 17 22 13 23 33 - electrical connections Patent Application Publication Apr. 29, 2010 Sheet 1 of 33 US 2010/0105035 A1 Figure 1 ecolumescene High-field EL Light Emitting Diodes DC-ELD AC-ELD LED OLED (Inorganic) Powder Thin Powder Thin Small Polymer Film Film Molecule Wo HIC No HIC Planar (1983) Primarily Pioneer (1998) In display products Sharp(1980) discrete (Sanyo-kodak) development products (backlight) Denso(1999) devices (1G) luxell(1996) HP (Sony) (iFire Technol) * High Information Content Patent Application Publication Apr. 29, 2010 Sheet 2 of 33 US 2010/0105035 A1 Figure 2 Rear electrode Dielectric PhosphorITO film ICICICI OC NRWA ORON /E NECRA NE SVYWWE N ? Stric ARBON NY N. W EPANSPAR NE ry "Strastrong B OUTPUT LIGHT Patent Application Publication Apr. 29, 2010 Sheet 3 of 33 US 2010/0105035 A1 Figure 3 sy yeaktha is a Battery Power Patent Application Publication Apr. 29, 2010 Sheet 4 of 33 US 2010/0105035 A1 Figure 4A 14 33 = electrical connections Patent Application Publication Apr. 29, 2010 Sheet 5 of 33 US 2010/0105035 A1 Figure 4B B 10 33 = electrical connections Patent Application Publication Apr. 29, 2010 Sheet 6 of 33 US 2010/0105035 A1 Figure 5 27. ELLAMP 31. External visual display (LCD) 28. Excitation Filter 29. Emission Filter //IV //IV ACHS(YYYSV A A A V V V V 30. CMOS carthera 21. Microcontroller/ module processor Patent Application Publication Apr. 29, 2010 Sheet 7 of 33 US 2010/0105035 A1 Figure 6 Patent Application Publication Apr. 29, 2010 Sheet 8 of 33 US 2010/0105035 A1 1-chip un hand-held 14+ -ch ip uni hand-held 14-ch ip uni any thermal cycler Patent Application Publication Apr. 29, 2010 Sheet 9 of 33 US 2010/0105035 A1 Figure 8 SYBR filters C Electroluminescent Film (0.2mm thick) Low power consumption, PCR chip in combination with transparent low R Road integrated heater and peltier cooling unit Light weight. Low power consumption, Light weight; and MEMS based construction. Patent Application Publication Apr. 29, 2010 Sheet 10 of 33 US 2010/0105035 A1 Figure 9 300mm) Hy -- AX, LEAD BGE: -SE" O42 WDE. O.35' 0EEP status A (it 1X 9 mm) 0.010" (0.25mm) opean OVER LEAOS: oozi” (amm) OENSIONAL TOLERANCE: 650 mm) OR ESS: to c3 (to.8mm) 6.01 TO 12" (SoTO300 m): ecos (atismm) Patent Application Publication Apr. 29, 2010 Sheet 11 of 33 US 2010/0105035 A1 Figure 10 0.06 inch Patent Application Publication Apr. 29, 2010 Sheet 12 of 33 US 2010/0105035 A1 Figure 11 forward primer reverse mes - oligonucleotide hooks Pre-Amplification (room temperature) eVeSe r forward primer -- oligonucleotide hooks - During Amplification (elevated temperatures) Patent Application Publication Apr. 29, 2010 Sheet 13 of 33 US 2010/0105035 A1 Figure 12 Hand-held E component COSt 1. Temperature E Cycler E S 50 till 4 Tirne (min) 2. Chip positive results S1 OO synthesis and Visualization 3. Light Source 4. Reporting Patent Application Publication Apr. 29, 2010 Sheet 14 Of 33 US 2010/0105035 A1 Figure 13 Competition Sensitivity Quantitative Startup Sample ample Cost Wolume per reaction PCR-chip wif es SOO S0 to 3 to 500 handheld SO al PCR-chip wi No. S8,400 S0 to 3 to 500 S20 al S35,000 S18 100 Technologies RAZOR Bio-Seg. S31,500 S38 HANAA T. T. T. T. T. Biasetis Yes S70,000 S1,000 MM Source Yes Service S300 to to S Niolecular based S600 per (add S15S sample per target) Patent Application Publication Apr. 29, 2010 Sheet 15 of 33 US 2010/0105035 A1 Figure 14 900000 Bioyaries (M- ES) seles) 8OOOOO Conventional ORAR 7OOOOO 6OOOOO M 500000 Conventional 400000 ?y 3OOOOO gy. Idaho Tech. 200OOO s RAZARBioTrove g PCR-Chip w/ essessesseesear" Thernpig 'elp & hand-held S eS S 'S $ $$$$$$$$$$sSS SS (S No. of samples (50 targets each) Patent Application Publication Apr. 29, 2010 Sheet 16 of 33 US 2010/0105035 A1 Figure 15 OOOOOOO BioVaries (M-series) list Bio-Seeq (HANNA) OOOOOO Conventional QPCR I Conventional PCR RRRIdaho Tech. RAZAR BioTrove OOOOO PCR-Chip wif ThermoCycle PCR-Chip & hand-held S S S is SSSSSSSSSSSSSS (S (SSS S S S S SSSSSSSSSS No. of samples (60 targets each) Patent Application Publication Apr. 29, 2010 Sheet 17 of 33 US 2010/0105035 A1 Figure 16 BioVeri s M1M analyze Base Unit Consumable Price (U) Time of Samples # of Targets Sample volume Quantitatio n Sensitivity Portability (Weight) Patent Application Publication Apr. 29, 2010 Sheet 18 of 33 US 2010/0105035 A1 Figure 17 w Patent Application Publication Apr. 29, 2010 Sheet 19 of 33 US 2010/0105035 A1 Figure 18 Pass Pump required (tood, tasalskab, arrying withinario 80r Last basis retup deciftus Retire4 credic carries requirii uretter trava laser wisp a Purse and microtic at litterta East to get to the is totrisardaralysis chartress vispi off wise if lectic state twis aidered distra fire Seco so ARRoon riperature be careful rattatue dNA has to ve orotherCR clip CR chiparply ?pecific tauncretipoes Chip Rexacted Fan fested to cooking parts olthe DNA (schia: soft C issi) for trerare bacterial part arbi ristics distristoralia daress if sacca als) is recred controle cycles between reitsperature three Cerreraare: surfak (95 to 72 tas up in 40 devictoired times This can be tune in the PC darieg at ty, the CCO or hott sourco hockdates tapito ge crweis which coin UWitt sata is C Perforginists artiysts in vicina snapshot; ury Uttard odnopotassigst to Aiptures to erarie over the per chip air of urratt ?amplifications artication euros curwairadial dia Alex 4 fitaris cara rathoghis circycle (Pice tests with X cases in Airplied large frictoil cigangas) DNA skiigis Lopcir will for his parta parts, we no ior care cas Patent Application Publication Apr. 29, 2010 Sheet 20 of 33 US 2010/0105035 A1 Figure 19 e u fasci Patent Application Publication Apr. 29, 2010 Sheet 21 of 33 US 2010/0105035 A1 Figure 20 Control Chip as a { M s s s s s Patent Application Publication Apr. 29, 2010 Sheet 22 of 33 US 2010/0105035 A1 A Trehose 6 24 2. 34 1a 19 2.4 29 3.43 log target copy number) log targat oopy number 15 days 30 days ..a 24 29 3. s 2 25 3. B log target copy number) log?copy number) Patent Application Publication Apr. 29, 2010 Sheet 23 of 33 US 2010/0105035 A1 Figure 22 Targel is labeled with AGC AAA CGACCGGTGCGGAGTGCGA ACGGCCACGCCT Probe attached to chip surface at 3' end 55 OC 95 OC Patent Application Publication Apr. 29, 2010 Sheet 24 of 33 US 2010/0105035 A1 Figure 23 ABVEWale Solic State Relay Switch Termistor Data Acquisition Board N.USB 6009 doped silicon PCR chip Themistor D Times ec) Patent Application Publication Apr. 29, 2010 Sheet 25 of 33 US 2010/0105035 A1 Figure 24 +5V Supply from DAQ 10 Kohm Resistor Ground +SV Supply from DAQ Analog Output Ch0 from DAQ Analog Input Ch.3 on DAQ Therstor Resistor Analog Input Ch.4 on DAQ Patent Application Publication Apr. 29, 2010 Sheet 26 of 33 US 2010/0105035 A1 Figure 25 Patent Application Publication Apr. 29, 2010 Sheet 27 of 33 US 2010/0105035 A1 Figure 26 a firstiii. Patent Application Publication Apr. 29, 2010 Sheet 28 of 33 US 2010/0105035 A1 Figure 27 Electrophoresis RD reagents . Section : RD PCR PCR chamber/ reagents f chambor Sensor eater F teater Sensor Patent Application Publication Apr. 29, 2010 Sheet 29 of 33 US 2010/0105035 A1 Figure 28 (3) Sallurip for STR linics, sar (c) Stial fan to efiance colling Saye-ita is Pisabietoa Fo Spaktota S8Power SP-84 Accessible Pius Fur s. 2433-R250 Patent Application Publication Apr. 29, 2010 Sheet 30 of 33 US 2010/0105035 A1 Figure 29 Resequencing 10 bp 1x would cost S500 k.
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