METHOD AND DEVICE FOR PARTICLE ANALYSIS USING THERMOPHORESIS

• US 20110084218A1
• Filed: 05/20/2009
• Published: 04/14/2011
• Est. Priority Date: 05/20/2008
• Status: Active Grant

• Current Assignee: NanoTemper Technologies GmbH
• Original Assignee: Ludwig-Maximilians-Universitat Munchen
• Inventors: Jens Wienken, Christoph, Braun, Dieter, Duhr, Stefan, Baaske, Philipp
• Granted Patent: US 8,941,080 B2
• Time in Patent Office: Days
• US Class Current: 250/459.1
• CPC Class Codes: B01L 2300/041 Connecting closures to device or container
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 3/508 rigid containers not provided for above
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 2015/0038 Investigating nanoparticles
• CPC Class Codes: G01N 2015/1027 Determining speed or velocity of a particle
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 2021/1731 Temperature modulation
• CPC Class Codes: G01N 2035/00356 Holding samples at elevated temperature (incubation)
• CPC Class Codes: G01N 21/171 with calorimetric detection, e.g. with thermal lens detection
• CPC Class Codes: G01N 21/1717 with a modulation of one or more physical properties of the sample during the optical investigation, e.g. electro-reflectance
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence

Abstract

The present invention pertains to a device and method to measure thermo-optical, preferably thermophoretic, characteristics of particles in a solution. The method comprises the steps of: (a) providing a sample probe comprising marked particles in a solution; (b) providing a temperature control system for creating a temperature gradient within said sample probe by contact heating, electrical heating and/or cooling; (c) detecting the marked particles at a first time; (d) creating a temperature gradient within the sample probe by means of the temperature control system; (e) detecting the marked particles in the sample probe at a, preferably predetermined, second time and/or at a predetermined location within the temperature gradient, and (f) characterizing the particles based on said two detections.

Claims

1. Method to measure thermo-optical, preferably thermophoretic, characteristics of particles in a solution comprising the steps of:

(a) providing a sample probe comprising marked particles in a solution;

(b) providing a temperature control system for creating a temperature gradient within said sample probe by contact heating, electrical heating and/or cooling;

(c) detecting the marked particles at a first time;

(d) creating a temperature gradient within the sample probe by means of the temperature control system;

(e) detecting the marked particles in the sample probe at a, preferably predetermined, second time and/or at a predetermined location within the temperature gradient, and

(f) characterizing the particles based on said two detections.

2. Method according to claim 1, wherein the method comprises the step of performing a third detection at a, preferably predetermined, third time.

3. Method according to claim 1, wherein the second time and/or third time is while and/or after heating, particularly in order to measure back diffusion.

4. Method according to claim 1, wherein the first, second and/or third times are predetermined times, preferably depending on absolute time, the completion or conduction of further method steps, and/or relative time between the two times.

5. Method according to claim 1, further comprising the step of exciting luminescence, preferably fluorescence, of said marked particles wherein the detection steps comprise detecting luminescence, preferably fluorescence, of said excited particles.

6. Method according to claim 1, wherein the temperature control system controls the temperature by a heating means and/or cooling means using one or more of a wire, a Peltier element, a plate, a conductive path, means for creating a high frequency electric field in the sample probe, an indium-tin-oxide (ITO) element and/or means with a radiation absorbing surface.

7. Method according to claim 1, wherein detection is performed by use of one or more of epifluorescence (EPI) microscope, total internal reflection fluorescence (TIRF) microscope, confocal microscope, CCD, APD, PMT, and/or a microscope.

8. Method according to claim 1, wherein the temperature gradient lies in the range from about 0 K/μ

m to 20K/μ

m, preferably from 0.001 K/μ

m to 5K/μ

m, more preferably from 0.01K/μ

m to 1K/μ

m and more preferably is about 0.05K/μ

m.

9. Method according to claim 1, wherein the temperature increase in the sample probe lies in the range from about 0°

C. to 100°

C., preferably from 2°

C. to 50°

C. and more preferably from 5°

C. to 20°

C.

10. Method according to claim 1, wherein the sample probe volume lies in the range from about 1 μ

l to 100 μ

l, preferably from 0.1 μ

l to 30 μ

l and also preferably from 0.5 μ

l to 5 μ

l.

11. Method according to claim 1, wherein the time span between the first time and the second time lies in the range from about 1 μ

s to 60 min, preferably about 0.01 sec to 20 min, preferably about 1 sec to 10 min preferably 60 sec to 5 min.

12. Method according to claim 1, wherein the predetermined location within the temperature gradient is at the surface, the bottom, the side of a measurement chamber and/or the sample probe and/or at a predetermined distance from heating and/or cooling means or at a place of specific temperature, preferably close to the heating and/or cooling means or near a surface in a region of high temperature gradients.

13. Method according to claim 1, wherein a plurality of sample probes are measured subsequently and/or in parallel.

14. A device for measuring thermo-optical, preferably thermophoretic, characteristics of particles in a solution, in particular according to the method of claim 1, the device comprises:

a measurement chamber for receiving a sample probe containing marked particles in a solution;

means for detecting the marked particles in the sample probe;

a temperature control system for creating a temperature gradient within said sample probe by contact heating and/or cooling, and

a control means adapted for controlling said detection means to detect the marked particles at a first time and for controlling said detection means and temperature control system to detect the marked particles in the sample probe at a second time and/or at a predetermined location within a temperature gradient created by the temperature control system.

15. Device according to claim 14, wherein the control means is adapted to control the first and/or second times to be predetermined times, preferably depending on absolute time, the completion or conduction of further method steps and/or relative time between the two times.

16. Device according to claim 14, further comprising means for exciting luminescence, preferably fluorescence, of said marked particles wherein the detection means is adapted to detect luminescence, preferably fluorescence, of said excited particles.

17. Device according to claim 14, wherein the temperature control system comprises one or more of each or more of the following heating and/or cooling means:

a wire, a Peltier element, a plate, a conductive path, means for creating a high frequency electric field, an indium-tin-oxide (ITO) element and/or means with a radiation absorbing surface for controlling the temperature.

18. Device according to claim 14, wherein the detection means comprises one or more of epifluorescence (EPI) microscope, total internal reflection fluorescence (TIRF) microscope, confocal microscope, CCD, APD, PMT, and/or a microscope.

19. Device according to claim 14, wherein the device comprises means for characterizing the particles based on said detections.

20. Device according to claim 14, wherein the temperature control system is adapted to create a temperature gradient lying in the range from about 0K to 20K, preferably from 0.001 K/μ

m to 5K/μ

m, more preferably from 0.01K to 1K/μ

m and more preferably is about 0.05K.

21. Device according to claim 14, wherein the temperature control system is adapted to create a temperature rise in the sample probe lying in the range from about 0°

C. to 100°

C., preferably from 2°

C. to 50°

C. and more preferably from 5°

C. to 20°

C.

22. Device according to claim 14, wherein the device is adapted to receive a sample probe having a volume lying in the range from about 1 pl to 100 μ

l, preferably from 0.1 μ

l to 30 μ

l and also preferably from 0.5 μ

l to 5 μ

l.

23. Device according to claim 14, wherein control means is adapted to control the time span between the first time and the second time to lie in the range from about 1 μ

s to 60 min, preferably about 0.01 sec to 20 min, preferably about 1 sec to 10 min, preferably 60 sec to 5 min.

24. Device according to claim 14, wherein the detection means is adapted to conduct the detection at the surface, the bottom, the side of the measurement chamber or the sample probe and/or at a predetermined distance from heating means or at a place of specific temperature, preferably close to the heating means or near a surface in a region of high temperature gradients.

25. Device according to claim 14, wherein the device is adapted to measure a plurality of sample probes subsequently and/or in parallel.

26. Device according to claim 14, wherein the device comprises a substrate containing a measurement chamber for receiving the sample probe.

27. Device according to claim 14, wherein the measurement chamber and/or the sample probe is covered by a cover lid.

28. Device according to claim 27, wherein the cover lid comprises one or more filling holes and/or pin holes.

29. Device according to claim 14, wherein the measurement chamber is defined as a recess in a substrate.

30. Device according to claim 14, comprising filling holes for filling the measurement chamber.

31. Device according to claim 14, wherein one or more of the heating and/or cooling means of the temperature control system extend into and/or through the measurement chamber and/or the sample probe.

32. Device according to claim 14, wherein one or more of the heating and/or cooling means of the temperature control system contacts the measurement chamber and/or the sample probe.

33. Device according to claim 14, wherein one or more of the heating and/or cooling means are electrically isolated vis-à

-vis the measurement chamber and/or the sample probe, preferably by an electrical isolation coating and/or an electrical isolation layer.

34. Device according to claim 14, wherein the measurement chamber and/or the sample probe is covered by a cover lid and wherein the cover lid is a heating and/or cooling element, preferably an ITO element.

35. Device according to claim 14, wherein the measurement chamber is defined by a structured surface and/or the sample probe is positioned on a surface of a substrate as a droplet, and where the heating and/or cooling element is defined by/on/in the substrate on which the droplet is placed.

36. Device according to claim 14, wherein the temperature of the measurement chamber and/or the sample probe can be/is controlled in a way that biochemical reactions such as PCR can be carried out.

37. A method to measure thermo-optical, particularly thermophoretic, characteristics of particles in a solution by using a device according to claim 14.

38-41. -41. (canceled)