Simulation of normal fresh blood platelets for reference control
The present invention provides for novel analogs of fresh human platelets for use in hematological instrument. Methods for preparing such analogs are also described.
1. A process for the manufacture of human platelet analogs for use as reference controls in automated blood cell analyzers, the process comprising the steps of:
(a) shrinking red blood cells of a non-human vertebrate to the size of human platelets by extraction of cellular fluid from the red blood cells; and
(b) heating the cells from step (a) at a temperature of at least 50° C. for a time period of at least 1 day, wherein the cells are treated with a denaturing agent prior to the heating or at the same time of the heating.
2. The process of claim 1 , wherein step (a) further comprising heating the cells at a temperature of about 40° C. for about 0.5 to about 5 hours after the shrinking of the cells.
3. The process of claim 1 , wherein step (a) further comprising heating the cells at a temperature of about 40° C. for about 3 hours after the shrinking of the cells.
4. The process of claim 1 , wherein step (b) comprises treating the cells from step (a) with a denaturing agent for at least two hours and then removing the denaturing agent from the cells prior to the heating.
5. The process of claim 1 , further comprising step (c) fixing the cells from step (b) with a fixing agent.
6. The process of claim 5 , wherein the fixing agent is glutaraldehyde.
7. The process of claim 1 , wherein the red blood cells are goat red blood cells.
8. The process of claim 1 , wherein the heating in step (b) is performed at about 50° C. to about 75° C.
9. The process of claim 8 , wherein the heating in step (b) is performed at about 56° C.
10. The process of claim 1 , wherein the time period is at least 2 days.
11. The process of claim 1 , wherein the time period is at least 3 days.
12. The process of claim 1 , wherein the time period is at least 4 days.
13. The process of claim 1 , wherein the time period is at least 7 days.
14. The process of claim 1 , wherein the time period is at least 14 days.
15. The process of claim 1 , wherein the time period is about 21 days.
16. The process of claim 1 , wherein the denaturing agent is α-naphthol.
17. The process of claim 16 , wherein α-naphthol concentration is 100 g/L to 500 g/L.
18. The process of claim 16 , wherein α-naphthol concentration is at least 400 g/L.
19. The process of claim 16 , wherein α-naphthol concentration is at least 500 g/L.
20. The process of claim 16 , wherein the time period is at least 7 days and α-naphthol concentration is at least 400 g/L.
21. The process of claim 16 , wherein the time period is at least 14 days and α-naphthol concentration is at least 400 g/L.
22. The process of claim 16 , wherein the time period is at least 21 days and α-naphthol concentration is at least 400 g/L.
23. The process of claim 16 , wherein the time period is about 21 days and α-naphthol concentration is about 500 g/L.
24. Modified red blood cells for use as reference controls in automated blood cell analyzers prepared by the process of claim 1 .