Particle agglutination in a tip
View Patent ↗An apparatus and a related method for performing particle agglutination reactions in a single, disposable probe tip are disclosed. The probe tip includes a sample cavity for sample acquisition, at least one flanking cavity for the capture of particles by centrifugation or other means, a transition zone for the mixing of the sample with reagents for agglutination and a detection zone for the optical detection of particle agglutination. A mechanism may be attached to the probe tip for the controlled movement of fluids through the internal volume of the probe tip. The probe tip is particularly useful for the automation of high-throughput agglutination-type assays.
1. A probe tip for performing a visually detectable agglutination reaction after aspiration of reagents and sample therein, said probe tip comprising:
a) a first port configured to permit application of negative or positive pressure to an internal volume of said probe tip;
b) a sample cavity of said internal volume in fluid communication with said first port;
c) at least one flanking cavity of said internal volume disposed outwardly radial of said sample cavity and having a primary axis disposed at an acute angle relative to a vertical center axis of said probe tip, said at least one flanking cavity being shaped to capture particles in the sample after separation of the particles from the remainder of the sample, wherein said at least one flanking cavity is in fluid communication with said sample cavity;
d) a detection cavity of said internal volume configured for the detection of agglutinated particles within the sample, wherein said detection cavity is in fluid connection with said sample cavity;
e) a transition zone of said internal volume disposed between said sample cavity and said detection cavity, said transition zone being configured for rotational mixing of the sample moving back and forth through said transition zone between said detection cavity and said sample cavity through agitation thereof; and
f) a second port in fluid connection with said detection cavity, said second port being configured to permit reagents and sample to be aspirated into or dispensed from said internal volume of said probe tip.
2. The probe tip of claim 1 , wherein the sample comprises a cell suspension.
3. The probe tip of claim 2 , wherein the cell suspension comprises red blood cells.
4. The probe tip of claim 2 , wherein the cell suspension comprises a donor's red blood cells.
5. The probe tip of claim 2 , wherein the cell suspension comprises a patient's serum and a donor's red blood cells.
6. The probe tip of claim 1 , wherein the sample comprises a ligand-binding molecule.
7. The probe tip of claim 6 , wherein the ligand-binding molecule is an antibody.
8. The probe tip of claim 6 , wherein the ligand-binding molecule is Coomb's reagent.
9. The probe tip of claim 1 , wherein the particles are microspheres.
10. The probe tip of claim 9 , wherein the microspheres are bound to ligand-binding molecules.
11. The probe tip of claim 10 , wherein the ligand-binding molecules are antibodies.
12. The probe tip of claim 1 , wherein the particles are cells.
13. The probe tip of claim 1 , wherein the particles are red blood cells.
14. The probe tip of claim 1 , wherein the agglutination is hemagglutination.
15. The probe tip of claim 1 , wherein the vertical center axis of said sample cavity and the primary axis of said at least one flanking cavity form an angle of 45 degrees or less thereby creating an annular lip.
16. The probe tip of claim 1 , wherein the separation of the particles from the remainder of the sample results from separation by centrifugation.
17. The probe tip of claim 16 , wherein the centrifugation results from the rotation of the probe tip around said vertical center axis of said probe tip.
18. The probe tip of claim 1 , wherein said at least one flanking cavity is configured to permit the resuspension of the centrifuged particles with the reagents aspirated into said flanking cavity.
19. The probe tip of claim 1 , wherein said at flanking cavity is configured to permit the flow of the resuspended centrifuged particles into said sample and detection cavities of said probe tip.
20. The probe tip of claim 1 , wherein the internal diameter of the wall of said sample cavity is larger than the internal diameter of the wall of said detection cavity.
21. The probe tip of claim 1 , wherein the detection cavity is optically transparent to permit optical detection of agglutination.
22. The probe tip of claim 1 , wherein said probe tip is made from a material that permits detection of fluorescence or chemiluminescence in said detection cavity.
23. The probe tip of claim 1 , wherein the walls of said detection cavity are capable of transmitting light at certain pre-determined wavelengths.
24. A kit for performing visually detectable agglutination reactions in a single probe tip, said kit comprising:
a) a probe tip comprising:
i. a first port configured to permit negative or positive pressure to an internal volume of a probe tip;
ii. a sample cavity of said internal volume in fluid communication with said port;
iii. at least one flanking cavity of said internal volume being disposed radially outward in relation to said sample cavity relative to a center vertical axis of said probe tip, said at least one flanking cavity having a primary axis disposed at an acute angle relative to said center vertical axis of said probe tip, said at least one flanking cavity being configured to capture particles in the sample after separation of the particles from the remainder of the sample, wherein said at least one flanking cavity is in fluid communication with said sample cavity;
iv. a detection cavity of said internal volume configured for the detection of agglutination within the sample, wherein said detection cavity is in fluid connection with said sample cavity;
v. a transition zone of said internal volume between said sample cavity and said detection cavity, configured for rotational mixing of the sample moving back and forth through said transition zone; and
vi. a second port in fluid connection with said detection cavity, said second port configured to permit reagents to be aspirated into or dispensed from said internal volume of said probe tip, and
b) reagents for agglutination.
25. The kit of claim 24 , wherein said reagents for agglutination comprise one or more ligand-binding molecules.
26. The kit of claim 24 , wherein said reagents for agglutination comprise microspheres bound by ligand-binding molecules.
27. The kit of claim 25 , wherein said ligand binding molecules are antibodies.
28. The kit of claim 24 , wherein said reagents for agglutination comprise Coomb's reagent.
29. The kit of claim 24 , wherein said reagents for agglutination comprise red blood cells of known blood group.
30. The kit of claim 24 , wherein the particles are separated from the remainder of the sample by centrifugation or by separation in a magnetic field.