Device for isolating small polymeric beads from a suspension of such beads

• US 6,332,540 B1
• Filed: 12/02/1999
• Issued: 12/25/2001
• Est. Priority Date: 04/08/1997
• Status: Expired due to Fees

• Current Assignee: Smithkline Beecham PLC
• Original Assignee: Smithkline Beecham PLC
• Inventors: Paul, Frank, Payne, Richard C, James, Philip John
• Primary Examiner(s): Ellis, Christopher P.
• Assistant Examiner(s): RODRIGUEZ, JOSEPH C
• Application Number:
  US09/402,686
• Time in Patent Office: 754 Days
• Field of Search: 209/3.1, 209/3.2, 209/10, 209/552, 209/577, 209/579, 209/587, 406/86-88, 406/92, 406/94
• US Class Current: 209/3.1
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00468 by manipulation of individual beads
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties

Abstract

A device for separating polymeric beads from a suspension of such beads comprising a flow cell through which the suspension of beads flows, and a radiation source which the stream of beads crosses. A detector detects the presence of a bead in the flow cell and control means directs the bead to a bead outlet channel.

Claims

1. A device for separating polymeric beads from a suspension of a plurality of such beads in a suspension fluid being a liquid, which comprises:

a flow cell through which a suspension of the beads in the suspension fluid can flow;

the flow cell having an inlet end through which the suspension of beads may be introduced into the flow cell, such that the beads are arranged in a successive serial flow stream as they flow through the flow cell toward an exit end of the cell;

a radiation source capable of directing incident radiation into the flow cell in an incident direction such that as beads flow through the flow cell they intercept the incident radiation;

a radiation detector positioned relative to the radiation source and to the flow of beads such that a change in the radiation detected by the detector occurs as the bead intercepts the incident beam;

a bead outlet channel and a suspension fluid outlet channel located downstream from the point where the beads intercept the incident radiation, and being in communication with the exit end;

valve means which in a first configuration directs suspension fluid flow preferentially through the bead outlet channel rather than through the suspension fluid outlet channel, and alternatively which in a second configuration directs suspension fluid flow preferentially through the suspension fluid outlet channel rather than through the bead outlet channel, so that in the first valve configuration beads are caused to flow through the bead outlet channel;

control means responsive to a signal from the radiation detector resulting from bead intercepting the incident beam, and which directs the valve means into the first valve configuration so that a bead flows through the bead outlet channel, and after the bead is downstream of the valve means then directs the valve means into the second valve configuration;

and wherein the valve means comprise a valve plug, and a conforming valve sleeve into which leads the common outlet channel and out of which lead the bead outlet and fluid outlet channels, the valve plug being rotatably moveable within the valve sleeve, a “

Y”

shaped channel passing through the valve plug,

such that by rotation of the plug within the sleeve the common outlet channel can be brought into communication with the “

Y”

shaped channel to allow entry of fluid from the common outlet channel into one or other of the limbs or stem of the “

Y”

, and exit of fluid respectively through the stem or another limb and thence into the bead exit channel to thereby provide a first configuration of the valve,

and rotation of the plug causing a different limb or the stem to be brought into communication with the bead exit channel or the fluid exit channel, to thereby allow entry of fluid from the common outlet channel into the different limb or stem of the “

Y”

, and exit of fluid through the stem or another limb and thence into the fluid exit channel to thereby provide a first configuration of the valve,

the valve sleeve having a flushing fluid conduit through which flushing fluid may be introduced into the valve sleeve from the exterior of the valve sleeve, the valve plug having a flushing fluid channel passing through the plug which by relative rotation of the plug within the sleeve and be brought into contact with both the flushing fluid conduit and the bead outlet channel to thereby direct flushing fluid from the flushing fluid conduit into the bead to thereby direct a bead along the bead outlet channel.

2. Device according to claim 1 characterised in that the flow cell is in the form of a tube of dimensions across the direction of flow of the beads about 1.5-4 times the diameter of the beads.

3. Device according to claim 2 characterized in that a flow of a sheath fluid is introduced around the flow of suspension of beads flowing through the inlet end of the flow cell, the sheath fluid flowing in the same direction as the flow of beads.

4. Device according to claim 1 characterised in that a flow of a sheath fluid is introduced around the flow of suspension of beads flowing through the inlet end of the flow cell, the sheath fluid flowing in the same direction as the flow of beads.

5. Device according to claim 4 characterised in that the suspension fluid and sheath fluid flow through the flow cell in laminar flow.

6. Device according to claim 4 characterised in that upstream of the inlet end of the flow cell, an inlet port for the suspension of beads in suspension fluid is provided, leading to an inlet conduit being of cross section comparable to the diameter of the beads and terminating in an orifice of comparable size which opens into a sheath fluid inlet chamber of wider cross section than the orifice and surrounding the orifice, at its exit end the chamber being in communication with the inlet end of the flow cell.

7. Device according to claim 6 characterized in that the sheath chamber is in the form of a chamber which tapers from a wider upstream end toward a narrower downstream end, being coaxial with the direction of flow through the flow cell, and in communication with the flow cell at its narrow end.

8. Device according to claim 5 characterized in that upstream of the inlet end of the flow cell, an inlet port for the suspension of beads in suspension fluid is provided, leading to an inlet conduit being of cross section comparable to the diameter of the beads and terminating in an orifice of comparable size which opens into a sheath fluid inlet chamber of wider cross section than the orifice and surrounding the orifice, at its exit end the chamber being in communication with the inlet end of the flow cell.

9. Device according to claim 1 characterised in that the incident direction is substantially normal to the flow direction, with the incident radiation in the form of a narrow beam which is intercepted by the flowing beads.

10. Device according to claim 1 characterised in that a radiation detector is positioned relative to the radiation source and to the flow of beads such that the det ec tor is at a non zero angle to the incident direction.

11. Device according to claim 1 characterised in that the radiation detector is positioned relative to the radiation source and to the flow of beads such that the detector is in line with the incident direction, but on the opposite side of the flow of beads to the radiation source.

12. Device according to claim 8 characterised in that between the flow of beads and the detector there is a radiation obstructer of size about the same order of magnitude as the cross section of the incident beam s uch that incident radiation does not directly impinge upon the detector but instead is scattered by beads around the obstructer into the detector.

13. Device according to claim 11 wherein the radiation obstructer is in the form of a strip of opaque material, thin in relation to its width, which may be adjusted by rotation about its longitudinal axis so as to present a surface varying between its width and its thickness to the incident beam.

14. Device according to claim 12 wherein the radiation obstructer is in the form of a strip of opaque material, thin in relation to its width, which may be adjusted by rotation about its longitudinal axis so as to present a surface varying between its width and its thickness to the incident beam.

15. Device according to claim 1 characterized in that the exit end of the flow cell is in communication with a common outlet channel aligned in the direction of flow of the beads, and which divides at a downstream point into two or more channels, being respectively a bead outlet channel and a fluid outlet channel.

16. Device according to claim 1 characterised in that the valve means comprise a valve body, having one or more channels therethrough, the channels and the common outlet channel and bead outlet and fluid outlet channels being moveable relative to each other so that the common outlet channel can be brought into communication with either the bead outlet or fluid outl et channels so as to direct a bead along the bead outlet channel.

17. Device according to claim 16 characterized in that the bead outlet channel is provided with means to introduce a flow of a flushing fluid upstream of a bead, so that a bead may be flushed along the bead outlet channel.

18. Device according to claim 1 characterized in that the valve means comprise shutters situated adjacent the bead outlet channel and fluid outlet channel which are reciprocally moveable into respective “

closed” and

“

open”

positions in which flow through the bead outlet channel and fluid outlet channel is respectively restricted or is allowed.

19. Device according to claim 18 characterized in that the bead outlet channel is provided with means to introduce a flow of a flushing fluid upstream of a bead, so that a bead may be flushed along the bead outlet channel.

20. Device according to claim 15 characterized in that the bead outlet channel is provided with means to introduce a flow of a flushing fluid upstream of a bead, so that a bead may be flushed along the bead outlet channel.

21. Device according to claim 1, characterised in that fluid from the fluid exit channel is recycled back to the sheath fluid chamber.

22. A device according to claim 1 wherein the flushing fluid is directed into the bead outlet channel whilst the valve means is in its second configuration, and in this second configuration the flow of suspension fluid through the bead outlet channel is closed.

23. Device according to claim 22 characterized in that a flow of a sheath fluid is introduced around the flow of suspension of beads flowing through the inlet end of the flow cell, the sheath fluid flowing in the same direction as the flow of beads.

24. Device according to claim 22 characterized in that the flow cell is in the form of a tube of dimensions across the direction of flow of the beads about 1.5-4 times the diameter of the beads.

25. Device according to claim 22 characterized in that the radiation detector is positioned relative to the radiation source and to the flow of beads such that the detector is in line with the incident direction, but on the opposite side of the flow of beads to the radiation source.

26. A device for separating polymeric beads from a suspension of a plurality of such beads in a fluid being a liquid, which comprises:

a flow cell through which a suspension of the beads in the fluid can flow;

the flow cell having an inlet end through which the suspension of beads may be introduced into the flow cell, such that the beads are arranged in a successive serial flow stream as they flow through the flow cell toward an exit end of the cell;

a radiation source capable of directing incident radiation into the flow cell in an incident direction such that as beads flow through the flow cell they intercept the incident radiation;

a radiation detector positioned relative to the radiation source and to the flow of beads such that a change in the radiation detected by the detector occurs as the bead intercepts the incident beam;

a bead outlet channel and a fluid outlet channel located downstream from the point where the beads intercept the incident radiation, and being in communication with the exit end;

valve means which in a first configuration directs fluid flow preferentially through the bead outlet channel rather than through the fluid outlet channel, and alternatively which in a second configuration directs fluid flow preferentially through the fluid outlet channel rather than through the bead outlet channel, so that in the first valve configuration beads are caused to flow through the bead outlet channel;

control means responsive to a signal from the radiation detector resulting from bead intercepting the incident beam, and which directs the valve means into the first valve configuration so that a bead flows through the bead outlet channel, and after the bead is downstream of the valve means then directs the valve means into the second valve configuration;

and wherein the radiation detector is positioned relative to the radiation source and to the flow of beads such that the detector is in line with the incident direction, but on the opposite side of the flow of beads to the radiation source, and between the flow of beads and the detector there is a radiation obstructer of size about the same order of magnitude as the cross section of the incident beam such that incident radiation does not directly impinge upon the detector but instead is scattered by beads around the obstructer into the detector,

and wherein the radiation obstructer is in the form of a strip of opaque material, thin in relation to its width, which may be adjusted by rotation about its longitudinal axis so as to present a surface varying between its width and its thickness to the incident beam.

27. Device according to claim 26 characterized in that the flow cell is in the form of a tube of dimensions across the direction of flow of the beads about 1.5-4 times the diameter of the beads.

28. Device according to claim 26 characterized in that a flow of a sheath fluid is introduced around the flow of suspension of beads flowing through the inlet end of the flow cell, the sheath fluid flowing in the same direction as the flow of beads.

29. Device according to claim 26 characterized in that the exit end of the flow cell is in communication with a common outlet channel aligned in the direction of flow of the beads, and which divides at a downstream point into two or more channels, being respectively a bead outlet channel and a fluid outlet channel.

30. Device according to claim 26 characterized in that the valve means comprise a valve body, having one or more channels therethrough, the channels and the common outlet channel and bead outlet and fluid outlet channels being moveable relative to each other so that the common outlet channel can be brought into communication with either the bead outlet or fluid outlet channels so as to direct a bead along the bead outlet channel.

31. Device according to claim 26 characterized in that the valve means comprise shutters situated adjacent the bead outlet channel and fluid outlet channel which are reciprocally moveable into respective “

closed” and

“

open”

positions in which flow through the bead outlet channel and fluid outlet channel is respectively restricted or is allowed.

32. Device according to claim 26 characterized in that the bead outlet channel is provided with means to introduce a flow of a flushing fluid upstream of a bead, so that a bead may be flushed along the bead outlet channel.