PBMC separation systems and methods of use
The invention provides cartridges, systems and methods for the separation of a PBMC fraction from a blood sample. The cartridge comprises a valve which is configured to be actuated by physical or magnetic force. The system comprises a rotor for rotating a cartridge and an actuator to operate a valve. A sensor may be provided in the system to provide feedback on the separation of the sample in the cartridge. The method provides for loading a blood sample into a cartridge, rotating the cartridge, and opening or closing a valve on the cartridge. The disclosed cartridges, systems and methods may be used together; however, it is also envisioned that the systems and methods may utilize other cartridges and/or other systems.
1 . A fluidic cartridge for separation of blood by rotational force, the fluidic cartridge comprising:
an inlet region comprising a sample reservoir having an opening adapted to receive a blood sample;
a separation region comprising a separation reservoir and a density gradient media reservoir comprising a medium for separation of blood components upon application of rotational force;
a collection region;
a plurality of channels connecting the inlet region, the separation region, and the collection region and connecting the density gradient media reservoir to the separation reservoir; and
a plurality of valves wherein a valve of the plurality of valves is on each channel of the plurality of channels, wherein each valve of the plurality of valves comprises an empty valve cavity and a valve cover, said valve cover bonded to a bottom portion of the fluidic cartridge that is adhered to a top portion of the fluidic cartridge, said valve cover having a rigid portion and a flexible portion that forms a continuous seal beneath the valve cavity,
wherein the bottom portion of the fluidic cartridge has two bottom portion openings at each valve to form a passage from a fluidic channel through the valve cavity when the valve is in an open position,
wherein the valve is configured to be opened and closed repeatedly, and
wherein the valve is not a wax valve.
2 . The fluidic cartridge of claim 1 , wherein the plurality of channels comprise a hydrophilic or hydrophobic coating.
3 . The fluidic cartridge of claim 1 , wherein the plurality of channels is coated with heparin or other anticoagulant substance.
4 . The fluidic cartridge of claim 1 , wherein the flexible portion is a flexible membrane.
5 . The fluidic cartridge of claim 1 , wherein the fluidic cartridge further comprises a wash reservoir, a storage media reservoir, and a waste collection reservoir, wherein the wash reservoir, the storage media reservoir, and the waste collection reservoir are connected to the sample reservoir by the plurality of channels.
6 . The fluidic cartridge of claim 1 , wherein the collection region comprises a plasma collection reservoir and a PBMC collection reservoir.
7 . The fluidic cartridge of claim 6 , wherein the plurality of channels connect the plasma collection reservoir to the separation reservoir and connects the PBMC collection reservoir to the sample reservoir.
8 . The fluidic cartridge of claim 1 , wherein the plurality of channels connect the density gradient media reservoir to the separation reservoir at a branched section that is connected to the outer edge of the separation reservoir.
9 . The fluidic cartridge of claim 1 , wherein the plurality of channels connect the outer edge of the sample reservoir to the inner edge of the separation reservoir.
10 . The fluidic cartridge of claim 1 , wherein the fluidic cartridge comprises a plurality of capillary stop valves connected to the plurality of channels to prevent fluidic movement through the fluidic cartridge without centrifugal force.
11 . The fluidic cartridge of claim 1 , wherein the valve on each of the plurality of channels further comprises a valve body inside the valve cavity molded to the flexible portion of the valve cover.
12 . A fluidic cartridge for separation of blood by rotational force, the fluidic cartridge comprising:
an inlet region comprising a wash reservoir and a sample reservoir having an opening adapted to receive a blood sample;
a separation region comprising a separation reservoir and a density gradient media reservoir comprising a medium for separation of blood components upon application of rotational force;
a collection region comprising a plasma collection reservoir, and a PBMC collection reservoir;
a plurality of channels connecting the sample reservoir, the plasma collection reservoir, and the density gradient media reservoir to the separation reservoir, and connecting the PBMC collection reservoir, the wash reservoir, a storage media reservoir, and a waste collection reservoir to the sample reservoir; and
a plurality of valves wherein a valve of the plurality of valves is on each channel of the plurality of channels, wherein each valve of the plurality of valves comprises an empty valve cavity and a valve cover, said valve cover bonded to a top portion of the fluidic cartridge that is adhered to a bottom portion of the fluidic cartridge, said valve cover having a rigid portion and a flexible portion that forms a continuous seal above the valve cavity,
wherein the top portion of the fluidic cartridge has two top portion openings at each valve to form a passage from a fluidic channel through the valve cavity when the valve is in an open position,
wherein the valve is configured to be opened and closed repeatedly, and
wherein the valve is not a wax valve.
13 . The fluidic cartridge of claim 12 , wherein the flexible portion is a flexible membrane.
14 . The fluidic cartridge of claim 12 , wherein the plurality of channels connect the density gradient media reservoir to the separation reservoir at a branched section that is connected to the outer edge of the separation reservoir.
15 . The fluidic cartridge of claim 12 , wherein the plurality of channels connect the outer edge of the sample reservoir to the inner edge of the separation reservoir.
16 . The fluidic cartridge of claim 12 , wherein the fluidic cartridge comprises a plurality of capillary stop valves connected to the plurality of channels to prevent fluidic movement through the fluidic cartridge without centrifugal force.
17 . The fluidic cartridge of claim 12 , wherein the valve on each of the plurality of channels further comprises a valve body inside the valve cavity molded to the flexible portion of the valve cover.
18 . A fluidic cartridge for separation of blood by rotational force, the fluidic cartridge comprising:
an inlet region comprising a wash reservoir and a sample reservoir having an opening adapted to receive a blood sample;
a separation region comprising a separation reservoir and a density gradient media reservoir comprising a medium for separation of blood components upon application of rotational force;
a collection region comprising a plasma collection reservoir, and a PBMC collection reservoir;
a plurality of channels connecting the plasma collection reservoir and the density gradient media reservoir to the separation reservoir, connecting the PBMC collection reservoir, the wash reservoir, a storage media reservoir, and a waste collection reservoir to the sample reservoir, and connecting the outer edge of the sample reservoir to the inner edge of the separation reservoir; and
a plurality of valves wherein a valve of the plurality of valves is on each channel of the plurality of channels, wherein each valve of the plurality of valves comprises an empty valve cavity and a valve cover, said valve cover bonded to a bottom portion of the fluidic cartridge that is adhered to a top portion of the fluidic cartridge, said valve cover having a rigid portion and a flexible portion comprising a flexible membrane that forms a continuous seal beneath the valve cavity,
wherein the bottom portion of the fluidic cartridge has two bottom portion openings at each valve to form a passage from a fluidic channel through the valve cavity when the valve is in an open position,
wherein the valve is configured to be opened and closed repeatedly,
wherein the valve is not a wax valve, and
wherein the fluidic cartridge comprises a plurality of capillary stop valves connected to the plurality of channels to prevent fluidic movement through the fluidic cartridge without centrifugal force.
19 . The fluidic cartridge of claim 18 , wherein the plurality of channels connect the density gradient media reservoir to the separation reservoir at a section that is connected to the outer edge of the separation reservoir.
20 . The fluidic cartridge of claim 18 , wherein the valve on each of the plurality of channels further comprises a valve body inside the valve cavity molded to the flexible portion of the valve cover.