Thermo-Scientific patent 6,628,382

1. Thermo-Scientific patent6,628,382 “Liquid photometer using surface tension to contain sample”

2. Thermo-Scientific patent6,628,382 The NanoDrop is a simple, easy-to-use spectrophotometer for measuring DNA, RNA, and protein concentrations insmall volume samples. “Liquid photometer using surface tension to contain sample”

3. Spectrophotometers measure absorbance pathlength Beer’s Law dictates absorption rate proportional to number of particles in optical path At 260 nm Double-stranded DNA has extinction coefficient ε260 = 0.020 (ug/ml)-1cm-1 Single-stranded DNA and RNA have extinction coefficient ε260 = 0.025 (ug/ml)-1cm-1

4. Spectrophotometers measure absorbance pathlength Beer’s Law dictates absorption rate proportional to number of particles in optical path At 260 nm Double-stranded DNA has extinction coefficient ε260 = 0.020 (ug/ml)-1cm-1 Single-stranded DNA and RNA have extinction coefficient ε260 = 0.025 (ug/ml)-1cm-1 The absorbance A260 = ε260 x (path length) x (concentration) A260 A280 Ratio gives measurement of protein contamination

5. State of the field prior Samples (often >50 ul) added to cuvettes ~1 cm path-length McMillan et al‘402, 1990 oceanoptics.com Surface tension,internal reflectionof ~1 ul dropletbetween rigid fibers SpectroPippeter from Ocean Optics aspirates ~ 2 ul fluid into small channel for measurement

6. NanoDrop optical fibercarrying lightto detector pin anvil/pedestal permanent magnet allen head-cap screw sampleof ~ 1ul plunger optical fibercarrying lightfrom source solenoid (with actuator)

7. CLAIM 1 ELEMENTS AT ISSUE 1. A photometric or spectrophotometric apparatus wherein -a sample in the form of a liquid drop is contained by surface tension forces between two planar surfaces, one containing               --a photometric or spectrophotometric source and the other               --a photometric or spectrophotometric detector and   -an optical path is established through the sample between the two surfaces said apparatus comprising:  [A] first and second anvil surfaces   -at least one being moveable relative the other to any one of three positions;  [B]an adjustable sample loading position so selected that   -the at least one moveable surface and   -the other surface are so remotely spaced that               --a liquid drop can be placed on the first surface;  [C] an adjustable sample compressionposition so selected that   -the surfaces are               --opposed and               --substantially parallel and               --proximally spaced so that                     ---the liquid                           ----wets and                           ----spreads upon both surfaces;  [D] an adjustable sample measuringposition so selected that         -the opposed substantially parallel surfaces are spaced apart to               --pull the sample into a columnwherein --it is contained by the surface tension thereby                             ----providing an optical path for a                                 -----photometric or                                 -----spectrophotometric measurement.

8. NanoDrop fixed pin plunger 0.2-2 mm sample columnof path-lengthoptimized forsample 1 ul sample solenoid Measurementposition Loadingposition Compressionposition an adjustable sample compression position so selected that … the liquid wets and spreads upon both surfaces; an adjustable sample measuring position so selected that the opposed substantially parallel surfaces are spaced apart to pull the sample into a column … an adjustable sample loading position so selected that … a liquid drop can be placed on the first surface;

9. Spec-2-Go interchangeablepin 0-1 ul sample,depending on pin size 0.2-2 mm pin stop 1 ul sample Loadingposition Compression/Measurementposition

10. LITERAL INFRINGEMENT There is a one-to-one correspondence between the Spec-2-Go and theelements of Claim 1 of the ‘382 patent NanoDrop position 1 an adjustable sample loading position so selected that   -the at least one moveable surface and   -the other surface are so remotely spaced that               --a liquid drop can be placed on the first surface; Spec-2-Go position 1(open arms and sample loading to bothanvils) NanoDrop position 2 an adjustable sample compression position so selected that   -the surfaces are               --opposed and               --substantially parallel and               --proximally spaced so that                     ---the liquid                           ----wets and                           ----spreads upon both surfaces; Spec-2-Go position 2(compression/measuring position) NanoDrop position 3 an adjustable sample measuring position so selected that         -the opposed substantially parallel surfaces are spaced apart to               --pull the sample into a column wherein                     ---it is contained by the surface tension thereby                             ----providing an optical path for a                                 -----photometric or                                 -----spectrophotometric measurement.

11. an adjustable sample measuring position so selected … to pull the sample into a column COMPRESSION POSITION MEASUREMENT POSITION NANODROP SPEC-2-GO COMPRESSION/MEASUREMENT POSITION

12. “Adjustable position” • Preset freedom of movement:(A) Claim 1: “an adjustable sample compression position” (uses anvils as stops for the NanoDrop arm-movements) • (B) Claim 21: “an adjustable stop regulates the spacing between the two surfaces when the solenoid plunger is depressed”Movement during measurement: • (A) Claim 25: “adjusting the distance between those two surfaces such that the sample is drawn into a column”

13. DOE Claim Element: Three positions

14. DOE Claim Element: Sample compression position … the liquid [drop] wets and spreads upon both surfaces

15. CLAIM 1 1. A photometric or spectrophotometric apparatus wherein -a sample in the form of a liquid drop is contained by surface tension forces between two planar surfaces, one containing               --a photometric or spectrophotometric source and the other               --a photometric or spectrophotometric detector and   -an optical path is established through the sample between the two surfaces said apparatus comprising:  [A] first and second anvil surfaces   -at least one being moveable relative the other to any one of three positions;  [B] an adjustable sample loading position so selected that   -the at least one moveable surface and   -the other surface are so remotely spaced that               --a liquid drop can be placed on the first surface;  [C] an adjustable sample compression position so selected that   -the surfaces are               --opposed and               --substantially parallel and               --proximally spaced so that                     ---the liquid                           ----wets and                           ----spreads upon both surfaces;  [D] an adjustable sample measuring position so selected that         -the opposed substantially parallel surfaces are spaced apart to               --pull the sample into a column wherein --it is contained by the surface tension thereby                             ----providing an optical path for a                                 -----photometric or                                 -----spectrophotometric measurement.

16. DOE Claim Element: Sample compression position … the liquid [drop] wets and spreads upon both surfaces

17. DOE Claim Element: Sample measuring position … [surfaces] pull the sample into a column

18. Figures 3a, 3b, 3c, and 3d