Ex vivo flow cytometry method and device
The invention relates to a method for diagnosing a disease state mediated by pathogenic cells. The method comprises the steps of combining with an ex vivo patient sample a composition comprising a conjugate or complex of the general formula A b -X, wherein the group A b comprises a ligand that binds to the pathogenic cells and the group X comprises an imaging agent, and detecting the pathogenic cells that express a receptor for the ligand using flow cytometry.
1. A method of detecting an imaging agent bound to pathogenic cells comprising the steps of:
(i) combining 1) an ex vivo patient sample comprising the pathogenic cells collected from a patient, wherein the ex vivo patient sample is a body fluid of the patient, and 2) a composition comprising a conjugate of the general formula
A b -X
wherein the group A b comprises a ligand that binds to the pathogenic cells and the group X is the imaging agent, wherein the imaging agent emits a detectable signal,
wherein A b comprises a folate receptor binding ligand, and wherein the folate receptor binding ligand, binds to the folate receptor on the pathogenic cells; and
(ii) detecting the signal from the imaging agent bound to the pathogenic cells using flow cytometry,
wherein the method is capable of quantifying the pathogenic cells at about 1 to about 10 cells per two milliliters of the ex vivo patient sample.
2. The method of claim 1 wherein the body fluid is selected from the group consisting of spinal fluid, lymph fluid, urine, mucus, and blood.
3. The method of claim 1 wherein the pathogenic cells are cancer cells.
4. The method of claim 3 wherein the cancer cells are metastatic cancer cells.
5. The method of claim 1 wherein the imaging agent comprises a chromophore.
6. The method of claim 5 wherein the chromophore is a fluorescent chromophore.
7. The method of claim 6 wherein the chromophore is selected from the group consisting of fluorescein, OREGON GREEN®, rhodamine, phycoerythrin, TEXAS RED®, DYLIGHT®, and ALEXAFLUOR®.
8. The method of claim 6 wherein the fluorescent chromophore is group X of A b -X, and A b -X is selected from the group consisting of folate-fluorescein, folate-OREGON GREEN®, folate-rhodamine, folate-phycoerythrin, folate-cys-TEXAS RED®, folate-ALEXAFLUOR®, and folate-DYLIGHT®.
9. The method of claim 1 further comprising the step of quantitating the pathogenic cells in the ex vivo patient sample.
10. The method of claim 1 wherein said pathogenic cells are detected using multiphoton flow cytometry.
11. The method of claim 1 wherein A b is folate.