Flow-through drum centrifuge

• US 6,991,738 B1
• Filed: 10/13/2004
• Issued: 01/31/2006
• Est. Priority Date: 10/13/2004
• Status: Active Grant

• Current Assignee: University of Washing, VisionGate, Inc.
• Original Assignee: University of Washington
• Inventors: Nelson, Alan C., Fauver, Mark E.
• Primary Examiner(s): Reifsnyder, David A.
• Application Number:
  US10/964,513
• Time in Patent Office: 475 Days
• Field of Search: 210/770, 210/778, 210/780, 210/802, 210/783, 210/787, 210/803, 210/360.1, 210/393, 210/394, 210/396, 210/402, 210/771, 210/791, 210/380.1, 118/55, 118/58, 422/72, 422/101, 427/2.1, 427/2.13, 494/36, 494/37, 494/43, 494/58
• US Class Current: 210/771
• CPC Class Codes: B04B 1/02 without inserted separating walls
• CPC Class Codes: B04B 15/08 for ventilating or producing a vacuum in the centrifuge
• CPC Class Codes: B04B 2005/0478 with filters in the separation chamber
• CPC Class Codes: B04B 2005/0485 with a displaceable piston in the centrifuge chamber
• CPC Class Codes: B04B 5/0442 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: G01N 1/2813 Producing thin layers of samples on a substrate, e.g. smearing, spinning-on G01N1/30 takes precedence
• CPC Class Codes: G01N 2001/2846 Cytocentrifuge method

Abstract

A process includes coating a cylinder having an inner wall and a cylinder axis with a gel coating on the inner wall. Then a specimen mixture including solvent is made to flow through the cylinder while the cylinder is being continuously rotated. The specimen mixture is initially directed to flow along the cylinder axis and such that specimen particles from the specimen mixture are accelerated off the cylinder axis toward the inner wall, so as to form a film of specimen particles embedded into the gel coating.

Claims

1. A process for separation of specimen particles, the process comprising coating a cylinder having an inner wall and a cylinder axis with an optical gel coating on the inner wall by injecting an optical gel into the cylinder and centrifuqating the optical gel to produce a uniform coating of the optical gel on the inner wall, then flowing a specimen mixture including solvent through the cylinder while the cylinder is being continuously rotated, wherein the specimen mixture is initially directed to flow along the cylinder axis and such that specimen particles from the specimen mixture are accelerated off the cylinder axis toward the inner wall, so as to form a film of specimen particles embedded into the uniform coating of the optical gel, and ejecting the film of specimen particles embedded into the uniform coating of the optical gel using a plunger mechanism having a tip and a base where the tip has a diameter matched to that of the inner diameter of the film of biological cell specimen particles embedded into the gel, so that the film of specimen particles embedded into the uniform coating of the optical gel is not actuated by the plunger tip, but so that there is a gap left between the film of specimen particles embedded into the uniform coating of the optical gel and the plunger tip, and wherein the plunger base has its diameter matched to that of the inner diameter of the cylinder, so that the film of specimen particles embedded into the uniform coating of the optical gel is forced forward when the plunger mechanism base engages the inner wall of the cylinder.

2. The process of claim 1 further comprising the step of removing the solvent from the film of specimen particles embedded into the uniform coating of the optical gel.

3. The process of claim 1 wherein the film of specimen particles embedded into the uniform coating of the optical gel include specimens of interest in a separation order and wherein the plunger tip maintains the separation order during plunging.

4. The process of claim 1 further comprising the steps of injecting the film of specimen particles into a microcapillary tube and operating a selector valve to discard portions of the film of specimen particles having negligible specimen particles.

5. The process of claim 1 wherein the uniform coating of the optical gel has a thickness that ranges from 100 microns to 1 mm.

6. The process of claim 1 including the step of blowing compressed air through the cylinder to remove solvent before ejecting the film of specimen particles embedded into the uniform coating of the optical gel.

7. The process of claim 1 wherein the specimen mixture comprises a biological sample including a solvent and cell mixture.

8. A process for separation of specimen particles, the process comprising the steps of:

injecting optical gel into a centrifuge having an inner wall;

centrifuging the optical gel to produce a coating of optical gel on the inner wall;

flowing a specimen mixture including solvent through the centrifuge while the centrifuge is rotated such that specimen particles from the specimen mixture are accelerated toward the inner wall and embedded into the coating of optical gel;

removing solvent from the coating of optical gel and specimen particles to form a film of specimen particles in optical gel; and

ejecting the film of specimen particles in optical gel into a microcapillary tube by using a plunger mechanism having a tip and a base where the tip has a diameter matched to that of the inner diameter of the film of specimen particles in optical gel, so that the film of specimen particles in optical gel is not actuated by the plunger tip, but so that there is a gap left between the coating and the plunger tip, and wherein the plunger base is matched to engage the inner wall, so that the film of specimen particles in optical gel is forced forward when the base engages the inner wall of the centrifuge.

9. The process of claim 8 wherein the optical gel is selected from the group consisting of optical gels, oils, fluids, polymer and epoxy.

10. The process of claim 8 further comprising the step of operating a selector valve to discard portions of the film of specimen particles in optical gel having negligible specimen particles.

11. The process of claim 8 wherein the film of specimen particles in optical gel includes specimens of interest in a separation order and wherein the plunger tip maintains the separation order during plunging.

12. The process of claim 8 wherein the coating has a thickness that ranges from 100 microns to 1 mm.

13. The process of claim 8 wherein the specimen mixture comprises a biological sample including a solvent and cell mixture.

14. A process for separation of specimen particles, the process comprising the steps of:

coating a cylinder having an inner wall and a cylinder axis by injecting an optical gel into the cylinder and centrifugating the optical gel to produce a uniform coating of the optical gel on the inner wall, wherein the optical gel has a selected index of refraction matched for subsequent imaging of the specimen particles;

flowing a specimen mixture including solvent through the cylinder while the cylinder is being continuously rotated, wherein the specimen mixture is initially directed to flow along the cylinder axis and such that specimen particles from the specimen mixture are accelerated off the cylinder axis toward the inner wall, so as to form a film of specimen particles embedded into the uniform coating of the optical gel; and

ejecting the film of specimen particles embedded into the uniform coating of the optical gel from the cylinder into a microcapillary tube for imaging by an imaging system.

15. A flow-through centrifuge system comprising:

a centrifuge having a gel-coated inner surface;

a mixture container for holding a specimen mixture coupled to the centrifuge, wherein the specimen mixture is injected into the centrifuge and rotated to embed specimen particles into the gel-coated inner surface of the centrifuge to form a film of gel and specimen particles; and

a plunger mechanism having a tip and a base where the tip is sized so that there is a gap left between the film of gel and specimen particles and the tip when the tip is inserted into the centrifuge, and wherein the base is sized so that the film of gel and specimen particles is forced forward when the base engages the inner surface of the centrifuge.

16. The flow through centrifuge system of claim 15 further comprising:

a microcapillary tube coupled to receive the film of gel and specimen particles; and

a selector valve for selecting a section of the film of gel and specimen particles.

17. The system of claim 15 wherein the specimen mixture comprises a biological sample including a solvent and cell mixture.