Abstract:
A disposable kit for use in directing fluid through a biological cell separator device ( 10 ). The kit generally includes a separator tube ( 22 ), a buffer fluid container ( 34 ), cell sample container ( 32 ), separated cell container ( 60 ), and flushing fluid container ( 62 ), as well as various conduits ( 36, 38, 42, 50, 50   a,    50   b ) for connecting the containers ( 32, 34, 60, 62 ) and separator tube ( 22 ) in fluid communication together. A cell separator system is provided including a separator tube ( 22 ), magnet ( 20 ), pump ( 120 ) and a motorized drive unit ( 96 ). The motorized drive unit ( 96 ) is operatively connected to the magnet ( 20 ) to allow the magnet ( 20 ) to be moved a sufficient distance away from the separator tube ( 22 ) so as to allow cells adhered to the inside surface thereof to be flushed out of the tube ( 22 ).

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
       [0001]    This application is claims the benefit of priority of U.S. Provisional Patent Application Ser. No. 61/305,380, filed Feb. 17, 2010 (pending) and is related to the subject matter disclosed in U.S. application Ser. No. 12/514,618, filed on Dec. 11, 2009, the disclosures of which are fully incorporated by reference herein. 
     
    
     SUMMARY 
       [0002]    In one embodiment, a disposable kit is provided for directing a fluid sample of biological cells through a separator device and collecting separated cells from the sample. The kit includes a sample receiving container including an inlet and an outlet, the inlet coupled with an inlet conduit for receiving a buffer fluid and the outlet coupled with an outlet conduit. A separator tube is provided in the kit, and includes an inlet end and an outlet end The inlet end is coupled with the outlet conduit from the sample receiving container. The separator tube has a flow channel, which may be annular in cross sectional shape, for receiving a flow of the buffer fluid and sample. A separated cell container is further provided and includes an inlet port and a vacuum port. A separated cell conduit is coupled for fluid communication between the outlet end of the separator tube and the inlet port of the separated cell container. A flushing fluid container is also provided and includes an inlet port and a vacuum port. A flushing conduit is coupled for fluid communication between the outlet end of the separator tube and the inlet port of the flushing fluid container. 
         [0003]    The kit can further include a buffer fluid container coupled with the inlet conduit to the sample receiving container. At least a portion of the outlet conduit from the sample receiving container may comprise a flexible tubing for receipt within a first pinch valve. At least a portion of the separated cell conduit may comprise a flexible tubing for receipt within a second pinch valve. At least a portion of the flushing conduit may comprise a flexible tubing for receipt within a third pinch valve. A venting conduit may be connected for fluid communication with the flushing fluid container. A first pump conduit is coupled with the vacuum port of the separated cell container, and a second pump conduit is coupled with the vacuum port of the flushing fluid container. 
         [0004]    In another embodiment, a device is provided that separates a first group of biological cells from a mixture of at least first and second groups of cells in a suspending fluid. The device comprises a flow channel having a length, a magnet, a pump, and a motorized drive unit. The magnet is mounted along at least a portion of the length of the flow channel, such that a magnetic field is provided through at least a selected portion of the flow channel. The pump is operatively connected to the flow channel and is configured to direct the suspending fluid through the flow channel. The motorized drive unit is operatively coupled to the magnet and is adapted to move the magnet along the length of the flow channel to lessen the effect of the magnetic field on the selected portion of the flow channel. Preferably, the motorized drive unit is configured to move the magnet away from the flow channel by a distance sufficient to allow flushing of cells from an interior wall of the flow channel. 
         [0005]    Various additional features of the invention and the illustrative embodiments will become more apparent from a review of the following detailed description of the illustrative embodiments. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0006]      FIG. 1  is a perspective view of a cell separator device constructed according to an illustrative embodiment. 
           [0007]      FIG. 2  is a cross sectional view of the device shown in  FIG. 1 , taken along line  2 - 2  thereof. 
           [0008]      FIG. 3  is an enlarged perspective view of a portion of the device shown in  FIG. 1 , including the magnet and separated cell container. 
           [0009]      FIG. 4  is an exploded perspective view of the magnet and separator tube associated with the device of  FIG. 1 . 
           [0010]      FIG. 5  is an exploded perspective view of a portion of the magnet. 
           [0011]      FIG. 6  is a cross sectional view of the magnet and separator tube taken in a plane along the longitudinal axis thereof. 
           [0012]      FIG. 7  is a cross sectional view of the magnet and separator tube taken in a plane perpendicular to the longitudinal axis thereof. 
           [0013]      FIG. 8  is a perspective view of the disposable container and tubing system associated with the device of  FIG. 1 . 
       
    
    
     DETAILED DESCRIPTION 
       [0014]    Referring first to  FIGS. 1-3 , a device  10  is shown and includes a housing or support structure  12 . An electrical power plug  14  is provided for receiving a power cord to supply electric current to the various electric components associated with the device  10 . A main on/off power switch  16  is provided, as well as a control panel  18  for allowing the user to operate the device  10 . A magnet  20  receives a separator tube  22  for purposes to be discussed further below. The magnet  20  is supported by a support or mounting structure  30  for vertical movement, as also discussed below. 
         [0015]    A biological cell sample container  32  is provided and receives a sample of biological cells, such as a blood sample, to be processed in the device  10 . A buffer or suspension fluid container  34  receives a buffer fluid and is in fluid communication with the cell sample container  32  via an inlet conduit  36 . A venting conduit  38  is also in fluid communication with the buffer fluid container  34  and leads to a fitting  40  and a vent ( FIG. 8 ). An outlet conduit  42  of cell sample container  32  extends through a slot  44   a  in a pinch valve  44 . The various conduits associated with the device  10  are preferably formed of a highly flexible material, such as silicone, so that they may be opened and closed to fluid flow using the illustrated pinch valves, including pinch valve  44 , for reasons to be explained below. Conduit  42  leads to the inlet end  22   a  of the separator tube  22  as best shown in  FIG. 2 . 
         [0016]    Still referring to  FIGS. 1-3 , a conduit  50  is in fluid communication with the outlet end  22   b  of the separator tube  22  and splits into respective conduits  50   a  and  50   b.  Conduit  50   a  extends through a pinch valve  52 , while conduit  50   b  extends through pinch valve  54 . Conduits  50   a  and  50   b  are respectively directed into a separated cell container  60  and a flushing fluid container  62 . A pump conduit  64  communicates with the interior of the separated cell container  60  and leads to a positive displacement pump  120  ( FIG. 8 ) via a fitting  66 . Another pump conduit  68  leads to a fitting  70  and a second pump  122  ( FIG. 8 ). Pump conduit  68  communicates with the interior of the flushing fluid container  62 . 
         [0017]    Turning more specifically to  FIG. 2 , the magnet  20  is mounted for vertical, linear movement along respective rails  80  (only one shown in  FIG. 2 ) via linear motion elements  82  and mounting structure  84  affixed with the magnet  20 . A screw drive receiving assembly  86  is coupled with a screw drive element  90  such that when the screw drive element  90  is rotated by a motor  92 , the screw drive receiving assembly  86  and the attached mounting structure  84  and magnet  20  will move vertically along the screw drive element  90 . In this illustrative embodiment, therefore, a motorized drive unit  96  is formed generally by elements  82 ,  84 ,  86 ,  89  and  82 , Although not shown, the separator tube  22  may be suitably clipped or mounted at the top to the device  10  so that it remains stationary and stable relative to the magnet  20  and does not drop downwardly through the magnet  20 . 
         [0018]    Referring now to  FIGS. 4-7 , the magnet  20  comprises an assembly of various components including four outer, nonmagnetic (e.g., aluminum) plates  100   a,    100   b,    100   c,    100   d  that provide a casing for four magnet assemblies. Each magnet assembly, as best shown in  FIG. 7  comprises an elongate element  102  having a stepped configuration in cross section and formed of a magnetic material, such as iron. Element  102  receives a series of permanent magnet plate elements  104 ,  106 ,  108 ,  110 ,  112 . Upper and lower end caps  116 ,  118  are coupled with each assembly and are formed of nonmagnetic material, such as aluminum, to contain the magnetic field at opposite ends of the magnet  20 . 
       Operation 
       [0019]    The operation of the device  10  will now be described in connection with  FIGS. 1 ,  2  and  8 . A biological cell sample fluid is prepared and placed into the cell sample container  32 . This sample may be a blood sample prepared in the manner described by the above-incorporated patent application. The size of the sample may, for example, be 5 ml. Pinch valves  44  and  52  are each in an open condition allowing fluid flow through conduits  42  and  50 ,  50   a.  Pinch valve  54  is closed to prevent fluid flow past the pinch valve  54  in conduit  50   b.  The positive displacement pump  120  is operated at a flow rate of, for example, 5 ml/min. to apply vacuum to the system, including to the interior of the separated cell container  60  via the pump conduit  64 , conduits  50   a,    50  leading to the interior annular flow channel of the separator tube  22  as well as conduit  42 , sample cell container  32 , conduit  36 , and the buffer fluid container  34 . This applied vacuum draws buffer fluid into the cell sample container  32 . Approximately 30 ml of fluid is thereby collected in the separated cell container  60  by drawing the 5 ml sample and 25 ml of buffer fluid through the separator tube  22  and into the container  60 . During this initial draw of fluid using the positive displacement pump  120 , the magnetic cells are attracted and adhered to the interior wall of the separator tube  22  by the magnet  20  generally as discussed in the above-incorporated patent application. One difference between the magnet  20  and the previously disclosed magnet of the above-incorporated patent application is that magnet  20  develops a substantially uniform magnetic field along the length of the separator tube, as opposed to a magnetic field of gradually increasing magnitude. 
         [0020]    After the initial draw of 30 ml fluid, pinch valves  44  and  52  are closed and the system at least temporarily shuts down. If the user desires to operate the device  10  in a flushing mode, the magnet  20  is moved downwardly by the motorized drive unit so as to completely expose the separator tube  22 . This at least substantially entirely removes the magnetic field from the length of the separator tube  22 , however, the cells are still adhered to each other and to the inside wall of the tube. Pinch valves  44  and  54  are then opened, while pinch valve  52  is closed. The high flow diaphragm pump  122  is then activated at a flow rate of, for example, 200 ml/min. to draw additional buffer fluid through the system and flush or wash the cells adhered to the interior of the separator tube  22  out of the separator tube  22  and into the flushing fluid container  62  via conduit  50   b.  It will be understood that if the user only has interest in collecting the nonmagnetic cells initially collected in the separated cell container  60 , the user may choose not to perform the flushing step. The fluid and cells in the container  60 /or and  62  may be further processed and analyzed as desired. These containers  60  and  62 , as well as the other components, including the containers  32 ,  34 , silicone tubing and separator tube  22  shown in  FIG. 8  may comprise a disposable kit or system that is designed for one-time use with the device  10 . Thus, to avoid contamination from process-to-process, a new disposable kit or system, such as shown in  FIG. 8 , may be utilized with the device  10  each time a sample cell separation is performed. 
         [0021]    While the present invention has been illustrated by a description of various illustrative embodiments and while these embodiments have been described in some detail, it is not the intention of the Applicants to restrict or in any way limit the scope of the appended claims to such detail. Additional advantages and modifications will readily appear to those skilled in the art. The various features of the invention may be used alone or any combinations depending on the needs and preferences of the user. However, the invention itself should only be defined by the appended claims.