High temperature flow-through device for rapid solubilization and analysis

• US 7,592,139 B2
• Filed: 02/16/2005
• Issued: 09/22/2009
• Est. Priority Date: 09/24/2004
• Status: Expired due to Fees

• Current Assignee: National Technology & Engineering Solutions Of Sandia LLC (Honeywell International Inc.)
• Original Assignee: Sandia National Laboratories (Government of the United States of America)
• Inventors: Hukari, Kyle W., Patel, Kamlesh D., Peterson, Kenneth A., Renzi, Ronald F., West, Jason A. A.
• Primary Examiner(s): Chunduru; Suryaprabha
• Application Number:
  US11/059,079
• Publication Number:
  US 20090215023A1
• Time in Patent Office: 1,679 Days
• Field of Search: 435/287.2, 435/6, 435/287.1, 536/23.1, 536/22.1
• US Class Current: 435/6.11
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 7/00 Heating or cooling apparatus autoclaves B01J3/04; Heat insulating devices
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12M 47/20 Heating or cooling heating or cooling apparatus for laboratory uses B01L7/00
• CPC Class Codes: C12N 1/066 by physical methods

Abstract

Devices and methods for thermally lysing of biological material, for example vegetative bacterial cells and bacterial spores, are provided. Hot solution methods for solubilizing bacterial spores are described. Systems for direct analysis are disclosed including thermal lysers coupled to sample preparation stations. Integrated systems capable of performing sample lysis, labeling and protein fingerprint analysis of biological material, for example, vegetative bacterial cells, bacterial spores and viruses are provided.

Claims

1. A device for lysing biological material, comprising:

an inner conduit for transporting the biological material,

the inner conduit having an entrance and an exit,

the inner conduit being characterized as a capillary tube or a microchannel; and

a heater situated exterior to the inner conduit capable of heating the biological material to at least about 125°

C. while the biological material is transported within said inner conduit.

2. The device of claim 1, further comprising an outer conduit surrounding the inner conduit.

3. The device of claim 2, wherein said heater comprises a heating coil wrapped around the outer conduit.

4. The device of claim 1, wherein the microchannel resides in a first substrate layer and the heater comprises a metal trace formed on a second substrate layer, both the first and second substrate layers being bonded together.

5. The device of claim 1, wherein the heater is capable of heating the biological material to a temperature of at least about 200°

C.

6. A micro-total analysis system, comprising:

an analytical device, and

a system for delivering biological material to the analytical device, comprising;

a biological material source;

a thermal lyser comprising an entrance and an exit, the entrance being in fluid communication with the biological material source, the thermal lyser capable of heating the biological material to at least about 100°

C. while the biological material is transported within the thermal lyser;

a dye source in fluid communication with the exit of the thermal lyser;

a buffer source in fluid communication with the exit of the thermal lyser; and

an exit capable of being in fluid communication with the analytical device.

7. The micro-total analysis system of claim 6, wherein the analytical device is capable of analyzing biological material using electrophoresis.

8. A method of preparing a biological sample for analysis, comprising:

flowing a biological sample fluid comprising biological material and a solvent through a conduit;

heating the biological sample fluid as it flows through said conduit to greater than 100°

C. at a pressure greater than 1 atmosphere;

lysing at least a portion of the biological material; and

labeling at least a portion of the lysed biological material with a dye.

9. The method of claim 8, wherein the biological material includes bacterial cells, bacterial spores, viruses, animal cells, plant cells, or any combination thereof.

10. The method of claim 8, wherein the solvent comprises water, an organic solvent, or any combination thereof.

11. The method of claim 8, wherein the solvent consists essentially of water, ethylene glycol, or any combination thereof.

12. The method of claim 8, wherein the biological sample is heated to at least about 125°

C.

13. The method of claim 8, wherein the residence time of the biological material as it is heated is up to about 300 seconds.

14. The method of claim 8, further comprising at least one PCR step.

15. The method of claim 14, wherein the concentration of organic solvent present during the PCR step is less than at out 3%.

16. A method of analyzing a biological sample, comprising:

flowing a biological sample fluid comprising a biological material and a solvent at a pressure greater than 1 atmosphere;

heating the biological sample fluid to greater than 100°

C. as it flows;

lysing at least a portion of the biological material to release biomolecules;

labeling at least a portion of the biomolecules with a dye; and

measuring the molecular distribution of the labeled biomolecules.

17. The method of claim 16, wherein the solvent comprises water, an organic solvent, or any combination thereof.

18. The method of claim 17, wherein the solvent consists essentially of water, ethylene glycol, or any combination thereof.

19. The method of claim 17, wherein the molecular distribution is measured using capillary electrophoresis.

20. The method of claim 16, wherein the dye is a rapid reacting dye.