System for thermocycling of fluids in cartridges

• US 7,256,035 B1
• Filed: 09/02/2000
• Issued: 08/14/2007
• Est. Priority Date: 09/04/1999
• Status: Expired due to Fees

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Roche Molecular Systems (Roche Holding AG)
• Inventors: Sarofim, Emad, Elsenhans, Olivier, Schnell, Urban, Birrer, Lukas, Caratsch, Thomas
• Primary Examiner(s): Beisner; William H
• Application Number:
  US10/070,343
• Time in Patent Office: 2,537 Days
• Field of Search: 435/6, 435/91.2, 435/287.2, 435/288.7, 435/303.1, 422/68.1, 422/102, 422/82.09, 356/246
• US Class Current: 435/287.2
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 2035/00366 Several different temperatures used

Abstract

System for thermocycling of fluids in cartridges comprising a thermocycling unit in thermal contact with a first substantially planar and heat conducting wall of a cartridge, a light source for transmitting light into the interior of said cartridge through a second light transparent wall of said cartridge which is arranged substantially perpendicular to said first wall, a light detector for detecting light emerging from the interior of the cartridge through said second wall and a fluid providing unit coupled to an inlet of the cartridge for providing the cartridge with fluid and an outlet of the cartridge to drain gas when the cartridge is filled with fluid. The invention further concerns a cartridge for conducting the thermal cycling of fluids.

Claims

1. A system for thermocycling of fluids in cartridges comprising

a) a cartridge with at least one heat conducting wall and at least one light transparent wall, the cartridge having a fluid inlet and a fluid outlet which are connected by a channel, said cartridge having a protrusion into the channel, such that the channel is longer than the shortest distance between the fluid inlet and the fluid outlet to allow bubble-free flow-through filling of the cartridge, wherein the fluid inlet is directly connected to a top wall, a bottom wall, and a pair of sidewalls, wherein the side walls form an angle of 100°

-150°

with the walls of the fluid inlet, and wherein the fluid outlet is directly connected to the top wall, the bottom wall, and a pair of side walls, wherein the side walls form an angle of 100°

-150°

with the walls of the fluid outlet,

b) a thermocycling unit in thermal contact with said heat conducting wall of said cartridge,

c) a light source for transmitting light into the interior of said cartridge through said light transparent wall of said cartridge which is arranged substantially perpendicular to said heat conducting wall,

d) a light detector for detecting light emerging from the interior of the cartridge through said light transparent wall,

e) a fluid providing unit coupled to an inlet of the cartridge for providing the cartridge with fluid by flow-through-filling.

2. The system of claim 1, wherein the cartridge has a body comprising the light transparent wall and having at least one opening which is sealed by a foil providing said heat conducting wall.

3. The system of claim 2, wherein the body is a frame which is sealed by two foils providing heat conducting walls.

4. The system of claim 1 for conducting fluorescent measurements wherein the light detector detects fluorescent light emerging from the cartridge.

5. The system of claim 1, wherein the thermocycling unit comprises at least one plate in thermal contact with the heat conducting wall of the cartridge.

6. The system of claim 1, wherein the cartridge is wedge shaped and the thermocycling unit comprises a wedge shaped receiving section for receiving said wedge shaped cartridge.

7. The system of claim 6, wherein the opposing walls forming the wedge shaped cartridge comprise a top wall, and a bottom wall having an angle of 3°

to 8°

with respect to each other.

8. The system of claim 1, wherein said light transparent wall comprises a first section through which light is passed from the light source into the cartridge and a second section through which light is passed from the interior of the cartridge onto the light detector.

9. The system of claim 8, wherein said first section is tilted with respect to the illumination beam axis so that the light is refracted towards the second section of said transparent wall.

10. The system of claim 1 or 5, wherein said thermocycling unit comprises a plate in thermal contact with said heat conducting wall of the cartridge and which exerts pressure onto said wall.

11. The system of claim 1, wherein the quotient of the maximal width of said channel and the depth of said channel is in the range of 1 to 10.

12. The system of claim 1 or 11, wherein the depth of the channel is in the range of 0.5 to 5 mm.

13. A method for thermocycling of fluids employing a system according to claim 1, comprising the steps of filling the cartridge by a flow-through-process avoiding bubbles in a measuring section of the cartridge, thermal cycling of fluid in the cartridge, transmitting light into the cartridge and detecting light emerging the cartridge.

14. The method of claim 13 additionally comprising monitoring the light emerging the cartridge to monitor amplification of nucleic acids in the fluid during thermal cycling.

15. A cartridge for conducting thermal cycling of fluids, comprising

a) a substantially planar and heat conducting wall,

b) a light transparent wall which is disposed substantially vertical to said heat conducting wall,

c) a fluid inlet for providing the cartridge with fluid,

d) a fluid outlet for draining fluid or gas from the cartridge,

e) a channel connecting the fluid inlet and the fluid outlet, wherein said cartridge has a protrusion, into the channel, such that the channel between the fluid inlet and the fluid outlet is longer than the shortest distance between the fluid inlet and the fluid outlet to allow flow-through-filling of the cartridge to avoid bubbles in a measuring section of the cartridge, wherein the fluid inlet is directly connected to a top wall, a bottom wall, and a pair of side walls, wherein the side walls form an angle of 100°

-150°

with the walls of the fluid inlet, and wherein the fluid outlet is directly connected to the top wall, the bottom wall, and a pair of side walls, wherein the side walls form an angle of 100°

-150°

with the walls of the fluid outlet.

16. The cartridge of claim 15, wherein the top wall and the bottom wall are opposing heat conducting walls.

17. The cartridge of claim 15 or 16, wherein the heat conducting wall is a foil with a thickness of less than 200 μ

m.

18. The cartridge of claim 15 or 16, having opposing top and bottom walls from which at least one is a heat conducting wall and which form an angle of 3°

to 8°

with respect to each other.

19. The cartridge of claim 15, wherein the light transparent wall comprises a first section for transmitting light into the cartridge and a second section for transmitting light emerging from the cartridge.

20. The cartridge of claim 15, which is made from a body having at least one opening which is closed by a heat conducting foil.

21. The cartridge of claim 15, which is made from a frame which is closed by two opposing foils.

22. The cartridge of claim 15 having a thickness of 0.5 to 5 mm.

23. The cartridge of claim 15, wherein the quotient of the maximal width of said channel and the depth of said channel is in the range of 1 to 10.

24. The cartridge of claim 23, wherein the depth of the channel is in the range of 0.5 to 5 mm.