Source: https://patents.justia.com/patent/10488353
Timestamp: 2020-01-17 22:11:12
Document Index: 715983622

Matched Legal Cases: ['Application No. 61', 'Application No. 2013202808', 'Application No. 2013202808', 'Application No. 2', 'Application No. 2', 'Application No. 2', 'Application No. 16193348', 'Application No. 16193348']

US Patent for Apparatus and system for performing thermal melt analyses and amplifications Patent (Patent # 10,488,353 issued November 26, 2019) - Justia Patents Search
Justia Patents US Patent for Apparatus and system for performing thermal melt analyses and amplifications Patent (Patent # 10,488,353)
Feb 23, 2017 - GEN-PROBE INCORPORATED
This application is a divisional of U.S. application Ser. No. 13/956,060, filed Jul. 31, 2013, now U.S. Pat. No. 9,588,069, which claims the benefit of U.S. Provisional Application No. 61/677,790, filed Jul. 31, 2012, the contents of each of which applications is hereby incorporated by reference herein in its entirety.
Aspects of the invention are embodied in an apparatus configured to apply thermal energy to the contents of a receptacle to increase the temperature of the contents of the receptacle and to detect an optical signal emitted by the contents of the receptacle as the temperature of the contents is rising. The apparatus comprises a receptacle holder configured to receive and releasably hold a receptacle, a vessel-receiving thermal assembly including a portion thereof held at a constant elevated temperature relative to ambient temperature and configured to receive a portion of the receptacle and to apply thermal energy to the contents of the receptacle, a receptacle moving mechanism configured to effect relative movement between the receptacle holder and the vessel-receiving thermal assembly to place a portion of the receptacle held by the receptacle holder into the vessel-receiving thermal assembly and to remove the portion of the receptacle from the vessel-receiving thermal assembly; and an optical signal detecting device constructed and arranged to detect optical signals emitted by the contents of a receptacle held within the vessel-receiving thermal assembly while thermal energy is being applied to the contents by the vessel-receiving thermal assembly.
According to further aspects the receptacle holder comprises a cover positioned over a receptacle carried in the receptacle holder and a yoke comprising side walls along opposed sides of the yoke and lateral support flanges extending along bottom edges of the sides walls.
According to further aspects, the thermal element comprise a resistive foil covering at least a portion of the thermal block.
Further aspects of the invention are embodied in a system for performing a nucleic acid diagnostic assay on a sample carried within a receptacle. The system comprises a target isolation module configured to isolate a target nucleic acid within the sample and to separate the target nucleic acid from non-target components of the sample, an incubation module configured to incubate the contents of a receptacle and perform an amplification procedure on the separated target nucleic acid within the receptacle, a thermal melt analysis module configured to receive a receptacle and to increase the temperature of the contents of the receptacle from a first temperature to a second temperature and to detect and record an optical signal emitted by the contents of the receptacle while the temperature of the contents is rising from the first temperature to the second temperature. The thermal melt analysis module includes a thermal block maintained at a substantially constant temperature that is greater than the first temperature. The temperature of the contents of the receptacle is increased from the first temperature to the second temperature by placing a receptacle having contents that are initially at the first temperature into operative proximity to the thermal block so that heat energy from the thermal block increases the temperature of the contents of the receptacle from the first temperature to the second temperature. The system further includes a receptacle transport mechanism under computer control and configured to (1) provide a receptacle containing a sample to the target isolation module, (2) after the target nucleic acid has been separated from non-target components of the sample, remove the receptacle from the target isolation module, (3) after removing the receptacle from the target isolation module, provide the receptacle to the incubation module, (4) after the amplification procedure is complete, remove the receptacle from the incubation module, and (5) after removing the receptacle from the incubation module, provide the receptacle to the thermal melt analysis module.
Further aspects of the invention are embodied in a method for performing a thermal melt analysis within a thermal melt analysis module. The method comprises the steps of (a) maintaining a thermal block within the module at a steady-state temperature, (b) placing a receptacle within the module in thermal contact with the thermal block, wherein the receptacle has contents that are at an initial temperature that is lower than the steady-state temperature, (c) allowing the receptacle to dwell in thermal contact with the thermal block for at least a predetermined dwell period so that the temperature of the contents of the receptacle increases from the initial temperature to a temperature that is higher than the initial temperature, (d) while the temperature of the contents of the receptacle is increasing from the initial temperature to the temperature that is above the initial temperature, measuring an optical signal emitted from the contents of the receptacle, and (e) detecting a change in the measured optical signal as the temperature of the contents of the receptacle increases from the initial temperature to the temperature that is above the initial temperature.
According to further aspects, the steady-state temperature is between about 70° C. and about 120° C.
According to further aspects, the steady-state temperature is between about 70° C. and about 90° C.
According to further aspects, the change in the measured optical signal results from melting of the hydrogen bonds between hybridized nucleic acid sequences contained in the receptacle. Although nucleic acid melting is exemplified herein, the present apparatuses and methods are also useful in conducting melting analyses of a variety of polymers, including amino acid and nucleic acid based polymers such as polypeptides, proteins, and various length nucleic acid molecules.
Further aspects of the invention are embodied in a method for performing a thermal melt analysis of a sample within a steady-state temperature module without actively monitoring the temperature of the sample. The method comprises the steps of (a) maintaining a thermal block within the module at a steady-state temperature, (b) introducing the receptacle to the module, wherein the receptacle is placed in thermal contact with the thermal block, and wherein the receptacle has contents that are at an initial temperature that is lower than the steady-state temperature, (c) allowing the receptacle to dwell in thermal contact with the thermal block so that the temperature of the contents of the receptacle increases from the initial temperature to a temperature that is higher than the initial temperature, and measuring the elapsed time that the receptacle is in thermal contact with the thermal block, (d) while the temperature of the contents of the receptacle is increasing from the initial temperature to the temperature that is above the initial temperature, detecting an optical signal attributable to a calibrator present in the contents of the receptacle, wherein the calibrator generates a detectable signal when the calibrator is at a predetermined temperature, (e) measuring the elapsed time between introducing the receptacle to the module and the detection of the optical signal attributable to the calibrator, and (f) comparing the measured elapsed time between introducing the receptacle to the module and the detection of the optical signal attributable to the calibrator to a calibration curve to determine the temperature of the contents of the receptacle at any time while the receptacle is present in the module, wherein the calibration curve comprises a plot of time versus temperature.
The analyzer 100 further includes a receptacle transfer apparatus, which, in the illustrated embodiment, comprises a receptacle distributor 300, embodying aspects of the present invention. Each of the modules of the analyzer 100 includes a receptacle transfer portal through which receptacles are inserted into or removed from the respective modules. Each module may or may not include an openable door covering its receptacle portal. The receptacle distributor 300 is configured to move receptacles between the various modules and retrieve receptacles from the modules and deposit receptacles into the modules. In one embodiment, the receptacle distributor 300 includes a receptacle distribution head 312 configured to move in an X direction along a transport track assembly 458, rotate in a theta (Θ) direction, and move receptacles in an R direction into and out of the receptacle distribution head 312 and one of the modules of analyzer 100. An exemplary receptacle distributor is described in WO 2010/132885, entitled “Method and Apparatus for Effecting Transfer of Reaction Receptacles in an Instrument for Multi-Step Analytical Procedure,” the disclosure of which is hereby incorporated by reference.
Though they are not necessary to effectively practice the present methods, such calibrators often eliminate the need to actively monitor the temperature of the sample contained in the receptacle vessel. Such calibrators may be advantageous when, for example, different sample types, different ambient temperatures, different sample volumes, different receptacle vessel materials, different receptacle vessel wall thicknesses, different air gaps between the receptacle vessel and the heat source, and/or other sources of inter- or intra-sample variability are present to appropriately place individual melt profiles on a pre-determined, concurrently determined, or otherwise known melt curve.
Avg. R1 Avg. R2 Avg. R3 Avg. R4 Avg. R5 R Set 1 71.46° C. 71.50° C. 71.66° C. 71.58° C. 71.22° C. R Set 2 71.28° C. 71.78° C. 71.54° C. 71.58° C. 71.00° C. R Set 3 71.28° C. 71.46° C. 71.60° C. 71.50° C. 71.32° C. R Set 4 71.28° C. 71.44° C. 71.52° C. 71.46° C. 71.32° C. R Set 5 71.22° C. 71.68° C. 71.60° C. 71.58° C. 71.08° C. Standard 0.082 0.134 0.050 0.051 0.129 Deviation (SD) MEAN 71.30° C. 71.57° C. 71.58° C. 71.54° C. 71.18° C. ACTUAL 71.71° C. 71.71° C. 71.71° C. 71.71° C. 71.71° C. Difference −0.41 −0.14 −0.13 −0.17 −0.53
5 Cycle Average 71.43° C. (all cycles, receptacles, and locations) SD 0.19
Target HCV-4H; HCV-3B HCV-2B HCV-5A HCV-1A Receptacle R1 R2 R3 R4 R5 R set 1 72.80° C. 75.00° C. 71.26° C. 72.32° C. 79.04° C. R set 2 72.76° C. 75.10° C. 71.34° C. 72.48° C. 79.00° C. R set 3 72.60° C. 75.00° C. 71.20° C. 72.22° C. 78.86° C. R set 4 72.66° C. 75.00° C. 71.30° C. 72.26° C. 78.92° C. SD 0.08° C. 0.04° C. 0.05° C. 0.10° C. 0.07° C. MEAN 72.70° C. 75.02° C. 71.27° C. 72.32° C. 78.95° C. ACTUAL 73.61° C. 76.42° C. 71.71° C. 72.98° C. 80.2° C. Difference −0.91 −1.40 −0.44 −0.66 −1.25 R1 R2 R3 R4 R5 5 Cycle Avg. 72.70° C. 75.02° C. 71.27° C. 72.32° C. 78.95° C.
Target HCV-4H HCV-3B HCV-2B HCV-5A HCV-1A Receptacle R1 R2 R3 R4 R5 R set 1 72.94° C. 74.98° C. 71.28° C. 72.34° C. 78.98° C. R set 2 72.96° C. 75.26° C. 71.62° C. 72.76° C. 79.44° C. R set 3 73.10° C. 75.62° C. 71.86° C. 72.92° C. 79.34° C. R set 4 72.58° C. 75.54° C. 71.74° C. 72.64° C. 78.98° C. MEAN 10% 72.89° C. 75.35° C. 71.62° C. 72.66° C. 79.18° C. MEAN 100% 72.70° C. 75.02° C. 71.27° C. 72.32° C. 78.95° C. Difference 0.19 0.33 0.35 0.35 0.23 SD 10% 0.193 0.251 0.216 0.213 0.210 SD 100% 0.08° C. 0.04° C. 0.05° C. 0.10° C. 0.07° C. Scale Factor 2.42 5.82 4.19 2.15 3.00 R1 R2 R3 R4 R5 5 Cycle Avg. 72.89 75.35 71.62 72.66 79.18
1. A system for performing a thermal melt analysis, the system comprising:
a thermal melt analysis module comprising: a receptacle holder configured to receive and releasably hold a receptacle; a vessel-receiving thermal assembly configured to receive the receptacle and comprising a thermal block assembly for applying thermal energy to the contents of the receptacle; a receptacle elevator configured to lower and raise the receptacle holder relative to the vessel-receiving thermal assembly to thereby place the receptacle into the vessel-receiving thermal assembly and to remove the receptacle from the vessel-receiving thermal assembly, respectively; and an optical signal detecting device configured to detect optical signals emitted by the contents of a receptacle placed into the vessel-receiving thermal assembly while thermal energy is being applied to the contents by the vessel-receiving thermal assembly; and
a system controller programmed to control the thermal melt analysis module to perform the thermal melt analysis by: directing the receptacle elevator to lower the receptacle holder, thereby placing a receptacle held by the receptacle holder into the vessel-receiving thermal assembly; directing the thermal block assembly to apply thermal energy to the contents of the receptacle, such that the thermal block assembly is maintained at a steady-state temperature, wherein the steady-state temperature is greater than ambient temperature; and while the thermal block assembly is maintained at the steady-state temperature, directing the optical signal detecting device to measure an optical signal emitted from the contents of the receptacle as the temperature of the contents of the receptacle increases.
2. The system of claim 1, wherein the receptacle holder comprises:
a yoke comprising side walls along opposed sides of the yoke and lateral support flanges extending along bottom edges of the side walls.
3. The system of claim 1, wherein the thermal melt analysis module further comprises a receptacle present detector configured to detect the presence of a receptacle in the receptacle holder.
4. The system of claim 1, wherein the vessel-receiving thermal assembly further comprises a vessel alignment block, wherein the vessel alignment block is configured to position a portion of a receptacle carried by the receptacle holder into the thermal block assembly when the receptacle elevator effects relative movement between the receptacle holder and the vessel-receiving thermal assembly.
5. The system of claim 4, wherein the thermal melt analysis module further comprises a thermal element in thermal contact with the thermal block assembly.
6. The system of claim 5, wherein the thermal element comprises a resistive foil covering at least a portion of the thermal block assembly.
the thermal block assembly is formed from a thermally conductive material and comprises a receptacle opening formed therein, wherein the thermal block assembly is positioned with respect to the vessel alignment block so that the receptacle opening formed in the thermal block assembly is aligned with the alignment opening formed in the vessel alignment block so that a receptacle inserted through the alignment opening formed in the vessel alignment block is positioned within the receptacle opening formed in the thermal block assembly.
8. The system of claim 4, further comprising at least one signal hole formed in the thermal block assembly and extending into the receptacle opening formed therein, the signal hole being configured to enable the optical signal detecting device to detect optical signals emitted by the contents of a receptacle positioned within the receptacle opening.
9. The system of claim 7, wherein the thermal melt analysis module further comprises an interface block disposed between the vessel alignment block and the thermal block assembly and having an opening aligned with the alignment opening of the vessel alignment block and the receptacle opening of the thermal block assembly.
10. The system of claim 7, wherein the alignment opening formed in the vessel alignment block is circular in cross-section and the receptacle opening formed in the thermal block assembly is circular in cross-section.
11. The system of claim 4, wherein the vessel alignment block comprises a raised center portion extending across a top surface of the vessel alignment block and defining recess shoulder portions on opposite sides of the raised center portion.
12. The system of claim 4, wherein the thermal block assembly comprises one or more receptacle holes formed therein from a top surface of the thermal block assembly and a hollowed-out portion extending from a lower surface of the block and surrounding the one or more receptacle holes without extending into any of the receptacle holes.
13. The system of claim 12, wherein the thermal melt analysis module further comprises a bottom cover secured to a bottom surface of the thermal block assembly to substantially enclose the hollowed-out portion.
14. The system of claim 13, wherein the thermal melt analysis module further comprises signal holes formed in the thermal block assembly and the bottom cover and extending into the receptacle holes formed in the thermal block assembly, the signal holes being configured to enable the optical signal detecting device to detect optical signals emitted by the contents of receptacles positioned within the receptacle holes.
15. The system of claim 4, wherein the vessel alignment block includes one or more mounting blocks raised from a surface thereof at which the vessel alignment block is attached to the thermal block assembly.
the receptacle holder is configured to receive and releasably hold a plurality of receptacles;
the vessel-receiving thermal assembly is configured to receive a portion of a plurality of receptacles and to apply thermal energy to the contents of the receptacles; and
the thermal melt analysis module further comprises a detector translating mechanism configured to move the optical signal detecting device with respect to the vessel-receiving assembly to selectively position a signal detecting channel of the signal detecting device in detecting alignment with two or more different receptacles held within the vessel-receiving thermal assembly.
17. The system of claim 1, wherein the receptacle elevator comprises:
18. The system of claim 17, wherein the thermal melt analysis module further comprises:
19. The system of claim 18, wherein each position sensor comprises a slotted optical sensor configured to be activated by a tab projecting from a portion of the receptacle holder.
20. The system of claim 17, wherein the screw follower is attached to a translating support bracket to which the receptacle holder is attached.
21. The system of claim 20, wherein the thermal melt analysis module further comprises one or more isolation mounts disposed between the translating support bracket and the receptacle holder, each isolation mount comprising:
a pin extending from the translating support bracket though an opening formed in the receptacle holder; and
a coil spring coaxially surrounding the pin.
22. The system of claim 1, wherein the receptacle does not physically contact the thermal block assembly.
23. The system of claim 1, wherein the optical signal detecting device is configured to detect optical signals at two or more distinct and distinguishable wavelengths.
24. The system of claim 23, wherein the optical signal detecting device is configured to detect optical signals at six (6) distinct and distinguishable wavelengths.
25. The system of claim 1, wherein the receptacle holder and the vessel-receiving thermal assembly are configured such that only a lower end of a receptacle held by the receptacle holder can be placed into the vessel-receiving thermal assembly.
26. The system of claim 1, wherein the thermal melt analysis module further comprises a signal detecting device moving mechanism configured to move the optical signal detecting device with respect to vessel-receiving thermal assembly.
27. The system of claim 26, wherein the optical signal detecting device comprises two or more channels, each channel being configured to detect an optical signal at a distinct and distinguishable wavelength, and wherein the signal detecting device moving mechanism is configured to sequentially position each channel relative to the receptacle to enable the signal detecting device to sequentially detect the wavelength corresponding to each channel.
28. The system of claim 26, wherein the signal detecting device moving mechanism comprises:
29. The system of claim 28, wherein the thermal melt analysis module further comprises:
30. The system of claim 29, wherein each position sensor comprises a slotted optical sensor configured to be activated by a tab projecting from a portion of the optical signal detecting device or the signal detecting device moving mechanism.
31. A system for performing a nucleic acid diagnostic assay on a sample carried within a receptacle, comprising:
a thermal melt analysis module configured to receive a receptacle and to apply thermal energy to the contents of the receptacle and to detect and record an optical signal emitted by the contents of the receptacle while thermal energy is being applied to the contents of the receptacle, wherein the thermal melt analysis module includes a thermal block, and wherein thermal energy is applied to the contents of the receptacle by placing a receptacle into operative proximity to the thermal block wherein the thermal melt analysis module is configured to hold the thermal block at a constant elevated temperature relative to ambient temperature for the duration of a thermal melt analysis; and
(3) after removing the receptacle from the target isolation module, provide the receptacle to the incubation module;
4021123 May 3, 1977 Atwood et al.
4851330 July 25, 1989 Kohne
5354663 October 11, 1994 Charm et al.
5482384 January 9, 1996 Lyle
5501963 March 26, 1996 Burckhardt
5686272 November 11, 1997 Marshall et al.
5840573 November 24, 1998 Fields
5998217 December 7, 1999 Rao et al.
6197520 March 6, 2001 Wittwer et al.
6248518 June 19, 2001 Parkhurst et al.
6391940 May 21, 2002 Blackwell et al.
6448066 September 10, 2002 Wheatcroft
6503711 January 7, 2003 Krull et al.
6635427 October 21, 2003 Wittwer et al.
7019267 March 28, 2006 Weinfield et al.
7255833 August 14, 2007 Chang et al.
7282333 October 16, 2007 Brow et al.
7319022 January 15, 2008 Mahoney et al.
7327459 February 5, 2008 Kim et al.
7348141 March 25, 2008 French et al.
7374885 May 20, 2008 Becker et al.
7381811 June 3, 2008 Weisburg et al.
7419786 September 2, 2008 Kurane et al.
7456281 November 25, 2008 Dujols
7462469 December 9, 2008 Bass et al.
7466908 December 16, 2008 Lem et al.
7482121 January 27, 2009 Sorge et al.
7485442 February 3, 2009 Afonina et al.
7541147 June 2, 2009 Marshall et al.
7547512 June 16, 2009 Peiris et al.
7549978 June 23, 2009 Carlson et al.
9588069 March 7, 2017 Opalsky et al.
20040014119 January 22, 2004 Itoh et al.
20050092643 May 5, 2005 Craven
20050202491 September 15, 2005 Nelson et al.
20060127906 June 15, 2006 Lee et al.
20070148677 June 28, 2007 Chagovetz et al.
20070172836 July 26, 2007 Exner et al.
20070184453 August 9, 2007 Sagner et al.
20080000892 January 3, 2008 Hirano et al.
20080089818 April 17, 2008 Ammann et al.
20090029877 January 29, 2009 Ammann et al.
20090155808 June 18, 2009 Hansen et al.
20100075312 March 25, 2010 Davies et al.
20100099581 April 22, 2010 Arciniegas
20100279276 November 4, 2010 Kacian
20100288395 November 18, 2010 Hagen et al.
20110236960 September 29, 2011 Bird et al.
20120221252 August 30, 2012 Heinz et al.
2549754 December 2006 CA
102422163 April 2012 CN
4214866 July 1993 DE
19964054 July 2001 DE
102007057651 June 2009 DE
0933132 August 1999 EP
1686190 August 2006 EP
5312813 November 1993 JP
2005010049 January 2005 JP
2002116202 April 2012 JP
9215853 September 1992 WO
9913976 March 1999 WO
0049557 August 2000 WO
0173399 October 2001 WO
2010/132885 November 2010 WO
Stratagene (Mx3000P™ Real-Time PCR System Instruction Manual, attached, Dec. 31, 2004).
Buchman et al., “Selective RNA Amplification: A Novel Method Using dUMP-containing Primers and Uracil DNA Glycosylase,” PCR Methods and Applications, Aug. 1993, pp. 28-31, vol. 3, No. 1, Cold Spring Harbor Laboratory Press, Woodbury, NY, US.
Idaho Technology Inc., HR1™ High Resolution Melter, Technical Users Manual, Rev-01 (2003).
Imboden et al., “Simultaneous Detection of DNA and RNA by Differential Polymerase Chain Reaction (DIFF-PCR)”, PCR Methods and Applications, Aug. 1993, pp. 23-27, vol. 3, No. 1, Cold Spring Harbor Laboratory Press, Woodbury, NY, US.
Mullis et al., “Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction”, Methods in Enzymology, 1987, pp. 335-350, vol. 155, Academic Press, Inc., Elsevier Inc., Philadelphia, PA, US.
Reed et al., “High-resolution DNA melting analysis for simple and efficient molecular diagnostics,” Pharmacogenomics, Jun. 2007, pp. 597-608, vol. 8, No. 6, Future Medicine Ltd, London, United Kingdom.
Stratagene, Mx3000P™ Real-Time PCR System, Instruction Manual (2004).
Walker et al. “Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system,” Proc. Natl. Acad. Sci., Jan. 1, 1992, pp. 392-396, vol. 89, No. 1, Proc. Natl. Acad. Sci, USA.
Walker et al. “Strand displacement amplification—an isothermal, in vitro DNA amplification technique,” Nucleic Acids Res., 1992, pp. 1691-1696, vol. 20, No. 7, Oxford University Press, Oxford, UK.
Zhang, Modern Molecular Biology Laboratory Manual, 2000, pp. 255-260, Science Publishers, USA.
APO Notice of Acceptance, Australian Patent Application No. 2013202808, dated Nov. 4, 2014.
APO Patent Examination Report No. 1, Australian Patent Application No. 2013202808, dated Mar. 3, 2014.
CIPO Office Action, Canadian Patent Application No. 2,879,720, dated Mar. 6, 2015.
CIPO, Exam Report, Canadian Patent Application No. 2,879,720, dated Jul. 23, 2015.
CIPO, Notice of Allowance, Canadian Patent Application No. 2,879,720, dated Aug. 28, 2015.
EPO Extended European Search Report, European Patent Application No. 16193348.6, dated Nov. 7, 2016.
EPO Communication pursuant to Article 94(3) EPC, European Patent Application No. 16193348.6, dated Feb. 26, 2019.
PCT Search Report, International Application No. PCT/US2013/053021, dated Dec. 13, 2013.
PCT Written Opinion, International Application No. PCT/US2013/053021, dated Dec. 13, 2013.
PCT International Preliminary Examination Report, International Application No. PCT/US2013/053021, dated Feb. 3, 2015.
USPTO Non-Final Office Action, U.S. Appl. No. 13/956,060, dated Mar. 19, 2015.
USPTO Final Office Action, U.S. Appl. No. 13/956,060, dated Oct. 15, 2015.
USPTO Non-Final Office Action, U.S. Appl. No. 13/956,060, dated Apr. 27, 2016.
USPTO Notice of Allowance, U.S. Appl. No. 13/956,060, dated Oct. 26, 2016.
EPO Communication Pursuant to Article 94(3) EPC, European Patent Application 16193348.6, dated Sep. 30, 2019.
Patent number: 10488353
Patent Publication Number: 20170160218
Inventors: David Opalsky (San Diego, CA), Norbert D. Hagen (Carlsbad, CA), Rolf Silbert (Del Mar, CA), Sean S. Chiu (Redmond, WA), Haitao Li (San Diego, CA)
Application Number: 15/440,806
International Classification: B01L 7/00 (20060101); G01N 25/04 (20060101); G01N 21/64 (20060101); B01L 9/06 (20060101);