JP2013226454A - Apparatus and method for separating and isolating component of biological fluid - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide device for separating and isolating components of a biological fluid.SOLUTION: A device 10 for separating and isolating components of a biological fluid comprises a container 48 for containing the fluid to be processed, a tube cap assembly 46 for closing the container while providing filling and extraction communication therewith, and a float assembly 130 disposed within the container for funneling and controlling biological fluid flow into an inverted domed shaped isolation chamber within the float and controlling the biological fluid flow out of the isolation chamber for effecting an encapsulation or a sealed isolation of at least one component or fraction of the biological fluid flow within the isolation chamber during a centrifugation process. The device further comprises a flexible tube 124 for connecting an extraction passageway disposed within the float assembly and an extraction valve of the tube cap assembly for allowing extraction of at least the one component or fraction encapsulated or isolated within the chamber.

Description

  The present invention generally relates to an apparatus and method for separating bodily fluids into components having different densities or concentrations, and more particularly, receiving bodily fluid samples, separating bodily fluid sample components, and bodily fluid component densities. Apparatus for isolating at least one target component from a non-target component of a body fluid sample based on a difference or concentration difference and extracting at least one target component for at least one diagnostic or therapeutic application And methods. The present invention is particularly useful in centrifuging blood and bone marrow into components.

  It is known to separate body fluids such as aspirated bone marrow or peripheral blood into their components, fractions, phases, or component layers (composition layers) by centrifugation. It is also known to provide a mechanical device composed of a tube (cylinder, tube) that accommodates a solid (solid content) separation device. This separation device is operated by centrifugal force, and has a difference in relative density. The body fluid is allowed to flow through or surrounding the piston, thereby separating the body fluid into one or more components above the solid separator or one or more components below the solid separator. For example, when the body fluid in the tube is blood, the centrifugation process causes red blood cells below the solid separation device, a low density layer of plasma above the solid separation device, and an intermediate density layer to a third density above the solid separation device. A buffy coat layer (soft layer) that defines a fraction of

  One of the earliest solids separators is disclosed in US Pat. No. 3,508,653 issued April 28, 1970 to Coleman. This device is an elastic cylinder made of rubber or other elastomer. A major problem with this device is that it is not possible to maintain a seal (sealing) because maintaining the precise inner diameter of the test tube when the mass is formed is costly. A subsequent improved solids separation apparatus is disclosed in U.S. Pat. No. 3,814,248 issued Jun. 4, 1974 to Lawhead. Next, in US Pat. No. 3,779,383 issued to Ayres on Dec. 18, 1973, the blood introduction end of the tube faces the movable separation device (movable separator) of the tube and is impossible. An apparatus is disclosed in contact with an permeable rubber plug. Following Ayres, issued on January 6, 1976, North, Jr. U.S. Pat. No. 3,931,018, which is used to separate serum and plasma using centrifugal force, and is inserted into a blood collection tube (blood collection tube) after blood collection A separation device is disclosed.

  In a patent granted to Levine et al. (U.S. Pat. No. 4,159,896 issued July 3, 1979), a solid separation device operating with centrifugal force is disclosed, in which the tube is A cylindrical float is disposed on the inside, and the float has an outer diameter that is precisely controlled to fit snugly within the bore (inner diameter) of the tube at rest. When used in collecting blood cells, the float is formed with an axial through hole that receives the leukocyte and platelet layers in the blood sample after centrifugation. The disclosed float is made from a plastic material having a specific gravity that floats in the concentrated red blood cells after centrifugation of the blood sample in the tube.

  Other patents to Levine et al. (US Pat. No. 5,393,674, issued February 28, 1995) have a cylindrical float that is large enough to process 1 ml of blood. A transparent plastic tube is disclosed which is provided and filled with a low-pressure inert gas. The float includes a through-hole, and is fixed and held in an initial position by making firm contact between the outside of the float and the inner wall of the tube before centrifugation. Unlike Coleman's invention, which includes a piston (or float) with no through-holes, Levine's float extends in the axial direction of the float body under centrifugation as a result of a large gravitational gradient from the top to the bottom of the float. (And subsequent lateral stenosis) results in a new position determined by that density relative to the density of the blood fraction. This large G force gradient (thousands of G) causes the tube to stretch and narrow so that it stretches and narrows when the rubber tube is pulled at both ends. The space between the exterior of the float and the interior of the tube that occurs during such centrifugation causes each blood component to move to a new location determined by its density relative to the float. Although not asserted in Levine, some of the redistributed blood components move through the holes during centrifugation, but the top and bottom of the through-holes are not closed, so they are wound up by centrifugation. Cells and platelets can easily enter red blood cells and plasma during normal processing after centrifugation. Designed primarily as a diagnostic tool performed by visual inspection of cells that at least temporarily occludes the through-hole immediately after centrifugation, Levine similarly collects these cells with an injection needle for additional diagnostic tests The possibility is disclosed. This collection method is inevitably inefficient as a means of cell recovery because the target cell (target cell) inevitably moves to the upper or lower part of the hole when the needle is inserted by invading the needle. It is.

  Thus, these known mechanical devices can generally separate body fluids into components or fractions. However, these devices are not very precise and the separated fractions are substantially mixed with each other, which is inefficient in separating body fluids into components or fractions. Furthermore, these known mechanical devices cannot easily and efficiently extract fractions other than the fraction at the top of the sample, especially the clinically important buffy coat fraction, and have a low recovery rate.

It is also known to provide more complex machinery to alleviate the above known problems. For example, the first example of the patent to Leach et al. (US Pat. No. 7,374,678 issued May 20, 2008) separates a blood-like sample into a plurality of fractions. An apparatus for disclosing is disclosed. Prior to centrifugation, the device includes a plunger (ie, a second piston) held in close proximity to a deformable top closed tube during centrifugation and a top closed deformable tube. It consists of a first piston (ie a buoy) fixed firmly in the vicinity, and under centrifugation of a blood sample, the wall of the tube is axially compressed and bent outwards, so that the buoy and the tube The buoys move downwardly between the inside of the wall and the tube that is lifted by the layer of red blood cells that is denser than the piston.
After centrifugation, the tube wall returns to its original size and the first piston is trapped in a new position that coincides with the interface position of the upper plasma fraction and the lower red blood cell fraction of the separated sample (capture). To do). Above or near the collection surface of this first piston (ie, buoy) is a third containing “small, unconcentrated, a specific amount of red blood cells, white blood cells, platelets and the buffy coat that occupies most of the blood sample”. There is a fraction. The device then uses a plunger (ie, a second piston) that is manually pushed down into the tube from an adjacent position at the top of the tube. The plunger (i.e., the second piston) includes a valve, which is set by a depth gauge, at a predetermined distance above the first piston that places the plunger at a distance away from the piston collection surface. The plasma is allowed to pass through the plunger while the plunger is below the depth of this, so that the bottom surface of the plunger (ie, the second piston) and the collecting surface of the first piston A third fraction is defined in between. The extraction of this third fraction is performed by a vacuum generated by a tube extending between a collection valve located in the top of the tube and a hole extending from the top of the plunger to the bottom of the plunger. Is called.

  Thus, this device relies on inaccurate axial compression and tube wall depressurization to control the flow path between each fraction, the centrifuge stops and the tube wall becomes its original It cannot contain fractions that are separated until after they return to size. Furthermore, the extraction of the third fraction requires the entry of the upper plasma. Therefore, this recently patented device is insufficient to separate bodily fluids into components or fractions and remains unable to solve the problem that the separated fractions are mixed together.

  As another example, Leach et al. Have a plunger (ie, a second piston) fixedly or slidably attached to a first piston or buoy, and these pairs are located inside a deformable tube with closed ends. The tube wall bends outward when centrifuging a sample such as blood, so that the pair moves toward the top of the tube and the pair and the tube wall Is lifted by a dense layer of red blood cells that flow downward. After centrifugation, the tube wall returns to its original position and grips around the first piston at the interface between the plasma fraction of the separated sample and most of the buffy coat. On or near the collection surface of this piston is “small, unconcentrated, a specific amount of red blood cells, white blood cells, platelets and the buffy coat that occupies most of the blood sample”. The extraction of the intermediate (buffy coat) to the third fraction consists of “interconnecting the cannula or perforated tube with the connection of the buoy cylinder” and connecting the extraction syringe to the cannula to connect the intermediate or third fraction. This is performed by generating a vacuum for extracting the gas from the space between the first piston and the second piston. In this example, only one centrifugal rotation is described, and the problem of inadequate separation of body fluid components or fractions and mixing of separated fractions cannot be alleviated. Moreover, in order to extract a fraction other than the upper fraction, it is necessary to enter at least one fraction other than the desired fraction to be extracted. In addition, this device relies on inaccurate axial compression and decompression of the tube wall to control the flow path between the fractions, until the centrifugation stops and the tube wall decompression is complete. It cannot contain separated fractions.

  Another problem associated with both embodiments, such as Leach, is that the buoy's collection surface, trough or water reservoir is at the desired density level of the target buffy coat fraction and together with the target red blood cells and platelets to be extracted. Must be shallow so as to prevent further accumulation of. Thus, this shallow trough causes target red blood cells and platelets to rest on the entire large surface area of the first piston, causing red blood cells and platelets to stick together, resulting in a final collection step. Efficiency is reduced. Another problem associated with both embodiments such as Leach et al. Is that it takes time and effort to attach and interconnect multiple parts to the device when performing the extraction process.

  In general, current processes for separating and extracting fractions from bodily fluids require many steps, which are time consuming and labor intensive, and have low recovery or recovery rates of target white blood cells and platelets. . Thus, providing a simplified and more efficient process that requires less time, effort and training to perform the process, and which has less red blood cell and platelet loss and can provide a positive economic impact. Is desired. Such a simplified process also allows for the setting of intra-operative settings by operating room nurses rather than remote laboratory settings by technicians, which allows patients to operate more quickly. Treatment is performed and sample mix-up can in principle be eliminated. Process simplification is also directly related to process reproducibility, which was a problem in the prior art.

  Thus, the prior art is based on the efficiency of recovering the cells of interest (target cells), the selectivity of the separation to reduce contaminating or non-target cells from the target cell population, and the extraction process which takes many steps, labor and time There are problems in many ways, including defects.

  Therefore, there is a need to overcome the major drawbacks of the prior art described in detail above.

  Accordingly, in one aspect, embodiments of the present invention facilitate fluid flow into the chamber of the funnel float and automatic self-sealing of this chamber to sequester at least one component or fraction of the body fluid being processed. To improve one or more major disadvantages of the known prior art by providing a separating and isolating device having a float assembly slidably enclosed in a container containing body fluid There is to do.

  In another aspect, embodiments of the present invention provide an inlet channel with a steep slope to a self-sealing isolation chamber so that components such as cells flowing into the isolation chamber It is possible to eliminate or prevent sticking on the surface of the entrance path with the mark.

  In one aspect, an embodiment of the present invention comprises a float that floats upstream and downstream in a circuit for fluid (body fluid) that passes across the float isolation chamber. The isolation chamber has a side valve, and the isolation chamber is responsive to a first differential pressure on the upstream valve and a second differential pressure on the downstream valve to isolate a target component of bodily fluid within the float. It is sealed by closing.

  In another aspect, another embodiment of the present invention provides an apparatus for separating and isolating body fluid components by centrifugation. The apparatus includes a storage container for storing bodily fluids having a large number of components, a float that is slidably disposed in the storage container and has an internal isolation chamber, and a first valve means on the first valve means. A first valve means for inflow of body fluid into the internal isolation chamber of the float in response to a first pressure difference of the body fluid, and a second valve means for second body fluid on the second valve means A second valve means for draining the body fluid from the float internal isolation chamber in response to the pressure differential, and isolating at least one target component of the body fluid within the float under centrifugation of the apparatus To isolate the interior of the chamber, the first valve means and the second valve means initially allow each component of the bodily fluid to flow through the interior isolation chamber, and then the first valve means In response to a first pressure differential fluid on Lube means, and the second in response to a second pressure differential of fluid on the valve means to seal the internal separation chamber closed.

  In another aspect, embodiments of the present invention provide an apparatus for separating and isolating bodily fluid components by centrifugation, the apparatus defining an interior chamber defining a body fluid containing a number of components. A storage container having a circumferential surface, slidably disposed inside the storage container and having a float that divides the storage container into a lower capacity area and an upper capacity area; The float has an inner isolation chamber defining an intermediate volume region and an outer circumferential surface spaced circumferentially from the inner circumferential surface of the storage container and defining a circumferential gap therebetween. A first valve means for opening and closing a body fluid communication (communication) or communication path (communication path) from the upper volume region to the inner isolation chamber inside the float, and from the inner isolation chamber inside the float Yong A second valve means for opening and closing the body fluid communication (communication) or the communication passage (communication path) to the lower volume region of the body, and at least of the many component body fluids inside the float inner isolation chamber In order to isolate one target component, under centrifugation, according to a first pressure difference of the body fluid on the first valve means and according to a second pressure difference of the body fluid on the second valve means From the upper volume region into the inner isolation chamber, exiting the inner isolation chamber into the lower volume region, exiting the lower volume region through the circumferential gap to the upper volume region, and back to the inner isolation chamber, at least The first valve means and the second valve means control the circulation of one closed-loop body fluid.

In another aspect, embodiments of the present invention provide a method for separating a bodily fluid having many mixed fractions using a conventional centrifuge, the method comprising a first fraction of bodily fluid. Centrifuging a device containing body fluid having a number of fractions, a second fraction and a third fraction to form a third fraction, and the interior of the float's inner isolation chamber during the centrifugation step Using a valve means for controlling the entry and exit of body fluid through the inner isolation chamber of the float of the apparatus. Further, and in one embodiment, the method further comprises the step of actuating a magnetic stir bar located inside the float's inner isolation chamber to agitate the third fraction after the centrifugation step. Do it. Also, and in one embodiment, the method further comprises collecting a third fraction of agitated body fluid from the device using an opening in open communication with the float's inner isolation chamber. In addition, and in one embodiment, the method includes the step of selectively adjusting the float to a density of about 1.02 grams / cm ³ to about 1.08 grams / cm ³ prior to the centrifugation step. The method further comprises connecting at least one weight (weight) to the float.

  In certain aspects, embodiments of the present invention provide an apparatus for separating and isolating components of bodily fluid by centrifugation, the apparatus comprising a closed bottom end, an open top end, and a closed bottom end and open. A container sidewall having an inner circumferential surface defining a containment chamber extending along a central longitudinal axis of the container, and a container sidewall having a container sidewall extending between A cap for selectively closing an open upper end of the container to define a storage container for containing a body fluid having the following components, a slidable arrangement in the container and a lower volume region and an upper side A float assembly that divides into volume regions, the float assembly being surrounded by an inner ceiling surface for defining a reverse dome-shaped chamber for isolating a target component of bodily fluid Bottom including the inner surface A lower cylindrical portion of the float that surrounds the cylindrical portion and the lower cylindrical portion and receives, directs, and facilitates (facilitates) the flow of body fluid from the upper volume region toward the inverted dome-shaped chamber Inside the float to provide open fluid communication between the upper cylindrical portion, including the conical upper surface, and a funnel-shaped cavity and an inverted dome-shaped chamber, defining a funnel-shaped cavity converging toward the And an open end exit passage disposed within the float to provide open communication between the open dome shaped chamber and the lower volume region. A first valve means for controlling fluid flow from the funnel-shaped cavity to the inverted dome-shaped chamber and fluid from the inverted dome-shaped chamber through the first valve means to the funnel-shaped cavity. Backflow A first valve means and a second valve means for selectively opening and closing an inlet passage open at an end in response to a first pressure difference on the first valve means, In order to control the outflow of fluid from the dome-shaped chamber to the lower volume region of the container and to prevent back flow of fluid from the lower volume region through the second valve means to the reverse dome-shaped chamber, the second valve And a second valve means for selectively opening and closing an outlet passage having an open end in accordance with a second pressure difference on the means, and a target component of body fluid during centrifugation The first valve means and the second valve means initially allow body fluid components to flow through the reverse dome-shaped chamber, Then, a first on the first valve means Depending on the pressure difference and on the second pressure difference on the second valve means, the inverted dome shaped chamber is closed to seal.

  Accordingly, it will be apparent that various modifications and applications may be made in accordance with the detailed description of the invention without departing from the scope and correct meaning of the claims set forth below.

1 is a perspective view of an embodiment of an apparatus for separating and isolating bodily fluid components. FIG. Figure 2 is a cross-sectional view of an embodiment of the main centrifuge tube of this device. FIG. 2 is an exploded view of the parts of an embodiment of the cap and valve assembly of the device. FIG. 4 is a top plan view of the cap illustrated in FIG. 3. FIG. 4 is a bottom plan view of the cap illustrated in FIG. 3. FIG. 2 is a cross-sectional view of the embodiment of the funnel float assembly illustrated in FIG. 1. FIG. 7 is a cross-sectional view of an embodiment of the float body of the funnel float assembly illustrated in FIG. 6. FIG. 8 is a top plan view of the outside of the float illustrated in FIG. 7. FIG. 8 is a bottom plan view of the outside of the float illustrated in FIG. 7. FIG. 7 is a perspective view of an embodiment of the funnel float cap of the funnel float assembly illustrated in FIG. 6. FIG. 11 is a top plan view of the funnel float cap illustrated in FIG. 10. FIG. 11 is a bottom plan view of the funnel float cap illustrated in FIG. 10. It is sectional drawing of the funnel float cap illustrated in FIG. It is a perspective view of the Example of an umbrella valve | bulb. It is a perspective view of the Example of a duckbill valve. It is a perspective view of the apparatus filled with body fluid. It is a perspective view of the apparatus before the centrifugation filled with body fluid. FIG. 3 is a cross-sectional view of the device after the centrifugation process. It is the perspective view which showed the target component extract | collected and isolated from the apparatus. 1 is a block diagram of conventional centrifugation and agitation used in the apparatus. It is a block diagram of the Example using an apparatus. FIG. 6 is a perspective view of another embodiment of an apparatus for separating and isolating bodily fluid components. It is a perspective view of the components which decomposed | disassembled the apparatus illustrated in FIG. FIG. 6 is a cross-sectional view of another embodiment of the main centrifuge tube. FIG. 6 is a cross-sectional view of another embodiment of a funnel float assembly. FIG. 26 is a perspective view of parts obtained by disassembling the funnel float assembly illustrated in FIG. 25. FIG. 26 is a cross-sectional view of an embodiment of the float body of the funnel float assembly illustrated in FIG. 25. It is a perspective view of the bottom part outside the float main body illustrated in FIG. FIG. 28 is a top plan view of the float application illustrated in FIG. 27. FIG. 26 is a perspective view of an embodiment of the funnel float cap of the funnel float assembly illustrated in FIG. 25. It is sectional drawing of the funnel float cap illustrated in FIG. FIG. 31 is a top plan view of the funnel float cap illustrated in FIG. 30. FIG. 31 is a bottom plan view of the funnel float cap illustrated in FIG. 30. FIG. 26 is a perspective view of the bottom cap of the funnel float assembly illustrated in FIG. 25. FIG. 35 is a cross-sectional view of the bottom cap illustrated in FIG. 34. FIG. 34 is a bottom plan view of the bottom cap illustrated in FIG. 33. FIG. 26 is a perspective view of another embodiment of a bottom cap for the funnel float assembly illustrated in FIG. 25. FIG. 38 is a cross-sectional view of the bottom cap illustrated in FIG. 37. FIG. 38 is a bottom plan view of the bottom cap illustrated in FIG. 37. FIG. 38 is a perspective view of an exploded part of a plurality of density adjustment weights (weights) and a bottom cap exemplified in FIG. 37. FIG. 38 is a perspective view of a state in which a plurality of density adjustment weights exemplified in FIG. 37 are coupled to a bottom cap. FIG. 35 is a perspective view of a state in which a plurality of weight mounts are added to the bottom cap illustrated in FIG. 34. FIG. 43 is a perspective view of a state in which a plurality of density adjustment weights are coupled to the bottom cap illustrated in FIG. 42. FIG. 7 is a perspective view of disassembled parts of a plurality of density adjustment weights and the float body of the funnel float assembly illustrated in FIG. 6.

  In each figure, like reference numerals in the various figures indicate like parts, reference numeral 10 represents an embodiment of a device for separating and isolating components of bodily fluids, and reference numeral 310 represents bodily fluids. Fig. 4 represents another embodiment of an apparatus for separating and isolating components.

  Referring to FIG. 1, in one embodiment, the apparatus 10 includes a container for defining a main centrifuge tube or container 12, a storage container 48 for receiving and containing a bodily fluid having multiple or many components. A tube cap assembly 46 for selectively closing the storage container into a first or lower volume region 260 below the float assembly 130 and a second or upper volume region 262 above the float assembly 130. The float assembly 130 to be divided, and the float assembly 130 and the tube cap assembly 46 are functionally connected to each other so as to perform a function, that is, the function of the tube cap assembly 46 by a crawling structure or a telescopic structure for moving up and down together with the float assembly 130 The flexible tube 124 is formed.

  Main centrifuge tube 12

  More particularly, with reference to FIGS. 1 and 2, the main centrifuge tube or vessel 12 has a closed bottom end 14, a substantially flat annular top 16 defining an open (open) top 18, And a cylindrical side wall 20 extending between the closed bottom end 14 and the annular top end 16. The cylindrical side wall 20 includes an outer cylindrical surface 22 and an inner circumferential or cylindrical surface 24 that defines a cylindrical storage chamber 26 extending along the central longitudinal axis 28 of the tube 124. This tube is also present in the central longitudinal axis of the device 10.

  In one embodiment, the closed bottom end 14 is formed as a substantially disk-shaped or disk-shaped member having an inner surface 30 and an outer surface 32. The inner surface 30 includes an inner rounded end that transitions into the inner cylindrical surface 24 of the cylindrical side wall 20. Similarly, the outer surface 32 includes an outer rounded end that transitions into the outer cylindrical surface 22 of the cylindrical sidewall 20. Further, the closed bottom end 14 may be selectively formed integrally with the cylindrical side wall 20 or configured to cover the side wall 20.

  Furthermore, the tube is preferably, but not limited to, made of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the tube 12 is constructed from a material such as, but not limited to, polycarbonate or polystyrene.

  Separating member 34

  With further reference to FIGS. 1 and 2, the tube 12 is comprised of a circular bottom end 36 that is coaxially disposed within the main centrifuge tube 12 and communicates or is integrally formed with the closed bottom end 14. A hollow cylindrical spacing member 34 having an open end (open), a circular upper end 38 defining an open (open) upper end 40, and extending between the circular bottom end 36 and the circular upper end 38. It further has an existing cylindrical side wall 42. The cylindrical side wall 42 includes an inner surface 44 that defines a cylindrical receiving chamber 45 disposed coaxially with the central longitudinal axis 28 of the main centrifuge tube 12. In one embodiment, the spacing member 34 has an outer diameter and height that are both substantially smaller than the outer diameter and height of the main centrifuge tube 12. The function or role of the separation member 34 will be described in detail below. Further, in one embodiment, the separation member 34 is composed of the same material as, but not limited to, the main centrifuge tube 12.

  Tube cap assembly 46

  With reference to FIG. 3, in one embodiment, the apparatus 10 is comprised of a tube cap 50, a hydrophobic air filter 80, an inlet valve 86, an outlet or extraction valve 100, and a barb fitting 114. A tube cap assembly 46 is further included.

  Tube cap 50

  With reference to FIGS. 3 to 5, the tube cap 50 extends from the upper surface 54 of the tube cap 50 to a lower annular portion 56 having a smaller diameter than the upper annular portion 52 and terminating at the lower surface 58 of the tube cap 50. It has an upper annular portion 52 that extends. Upper annular portion 52 extends adjacent to annular upper end 16 of cylindrical side wall 20, while lower annular portion 56 provides a connection between tube cap 50 and main centrifuge tube 12. The tube cap 50 fits into the open upper end 18 and extends to the cylindrical surface 24 inside the cylindrical side wall 20 to form an interference fit so that it remains in place within the upper end. Yes. In one embodiment, the tube cap 50 is medically bonded or joined to the main centrifuge tube 12.

  The tube cap 50 is also preferably, but not limited to, a material that is biocompatible and stable to gamma irradiation. In one embodiment, tube cap 50 is constructed from, but not limited to, a polycarbonate material.

  3 to 5, the tube cap 50 further includes a passage housing 60 that is attached to or integrally formed with the upper annular portion 52 of the tube cap 50. The passage housing 60 extends upward from the upper side surface 54 of the tube cap 50, and includes a housing branch port 66 and a housing inlet port 68 that is recessed and disposed in the lower side surface 58 of the tube cap 50. An L-shaped passage 62 having a first passage branch 66 extending there between. Similarly, the second passage branch 70 of the L-shaped extraction passage 62 extends between the passage branch bend 66 and a housing outlet port 72 disposed in the housing side wall 74 of the passage housing 60. . The second passage branch portion 70 extends away from the housing outlet port 72 and terminates before reaching the branch bent portion 66.

  The tube cap 50 is preferably, but not limited to, a material that is biocompatible and stable to gamma irradiation. In one embodiment, the tube cap 50 is composed of a material such as, but not limited to, polycarbonate or polystyrene.

  Further, the tube cap 50 is disposed adjacent to one side of the passage housing 60 and through the tube cap 50, and the tube cap 50 is disposed adjacent to the other side of the passage housing 60 and through the tube cap 50. And an air filter opening 78 formed therein.

  Air filter 80, inlet valve 86, extraction valve 100, barb joint 114

  With further reference to FIGS. 3-5, the tube cap assembly 46 is disposed within the air filter opening 78 and within the upper annular portion 52 of the tube cap 50 at a location surrounding the air filter opening 78. It has a hydrophobic air filter 80 located in a ledge 84 with a dent placed.

  The hydrophobic air filter 80 is preferably, but not limited to, a material that is biocompatible and stable to gamma irradiation. In one embodiment, the hydrophobic air filter 80 is comprised of a polypropylene material with, but not limited to, a PET filter material. As an example of the hydrophobic air filter 80, a part number X5009 manufactured by QoSina is commercially available.

  In addition, the tube cap assembly 46 has an inlet valve 86 that includes a conventional male luer lock head 88 having a male thread 90. The head 88 transitions into the shoulder 92 of the cylindrical body 94, which terminates in the lower side 96 and transitions into the lower tapered end 98. The lower tapered end 98 of the inlet valve 86 is disposed through the inlet valve opening 76 until the lower side 96 of the inlet valve 86 contacts the upper side 54 of the tube cap 50.

  In addition, the tube cap assembly 46 includes an outlet or extraction valve 100 that, in one embodiment, includes a conventional male luer lock head 102 having an external thread 104. The head 102 transitions into the shoulder 106 of the cylindrical body 108, which terminates at the lower side 110 and transitions into the lower tapered end 112. The lower tapered end 112 of the extraction valve 100 is disposed through the housing outlet port 72 until the lower side 110 of the extraction valve 100 contacts the housing sidewall 74 of the passage housing 60.

  Both the inlet valve 86 and the outlet or extraction valve 100 are preferably, but not limited to, materials that are biocompatible and stable to gamma irradiation. In one embodiment, both the inlet valve 86 and the outlet or extraction valve 100 are composed of polycarbonate material with, but not limited to, silicon rubber inserts. As an example of the inlet valve 86 and the outlet or extraction valve 100, part number 245501024 manufactured by Halkey-Roberts is commercially available.

  In addition, the tube cap assembly includes a barb fitting 114, which in one embodiment transitions from a short cylindrical portion 116, a short cylindrical portion 116 disposed within the housing input port 68, and the like. A flange 118 adjacent to a recessed shelf 120 disposed in the lower annular portion 56 at a position surrounding the housing input port 68 and a transition from the flange 118 and winding as shown in FIG. And an end 122 with a locking portion operably connected to the upper end 126 of the tube 124. The rolled tube 124, on the other hand, includes a lower end 128 connected to the float assembly 130, as described in detail below.

  Barb joint 114 is also preferably, but not limited to, made of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the barb joint 114 is comprised of, but not limited to, ABS material. An example of a barb joint 114 is Value Plastics, Inc. Commercially available as part number BDMR210-81.

  Float assembly 130

  With reference to FIGS. 1 and 6, as described above, the apparatus 10 includes a float assembly 130, which is for dual density subsurface funnel and isolation. As defined in the float assembly 130 and slidably disposed within the container 12, and as will be described in detail below, the container 12 is also connected to a first to lower volume region 260 below the float assembly 130. The second to upper volume region 262 above the float assembly 130 and the isolation or third volume region defined by the isolation chamber 208 in the float assembly 130.

  In one embodiment, the float assembly 130 includes a funnel and isolation float 132 comprised of a float body 134 and a float cap 194, a first check valve means in the form of an umbrella valve 234, and duckbill. It comprises a second check valve (check valve) means in the form of a valve 248. The funnel float 132 can be made as multiple or many elements or components, or as a single integrated element or component.

  Float body 134

  More specifically, with reference to FIGS. 6 and 7, the float body 134 includes a circular bottom surface 136, a frustum (conical) shaped top top surface 138, and between the circular bottom surface 136 and the frustum shaped top top surface 138. And a float side wall 140 extending to the surface. The circular bottom surface 136 can be substantially flat or taper (tapered) toward the central longitudinal axis 168 of the float body 134.

  The float sidewall 140 includes an outer circumferential or cylindrical surface 142 that extends between the circular outer edge 144 of the circular bottom surface 136 and the circular outer edge 146 of the frustum-shaped top top surface. The outer circumferential surface 142 of the float side wall 140 is the main centrifuge to define a circumferential gap 148 between the outer circumferential surface 142 of the float side wall 140 and the inner circumferential surface 24 of the tube 12. A diameter smaller than the diameter defined by the circumferential surface 24 inside the separation tube 12 is defined.

  In addition, the float sidewall 140 includes an inner hemispherical surface 150 that defines an inverted dome-shaped or hemispherical cavity 152 that extends from the rounded bottom section 154 of the hemispherical surface 150 into an inverted dome-shaped cavity. Extending to an upper annular shelf 156 defining an annular opening 158 in the cavity 152. The upper annular shelf 156 transitions into an inner cylindrical surface 160 that defines a cylindrical cavity 162 having an open end that surrounds the inverted dome-shaped cavity 152 so that the annular opening 158 is similarly cylindrical. An opening below the cavity 162 is defined. Similarly, the inner cylindrical surface 160 extends from the annular shelf 156 to the circular inner end 164 of the frustum-shaped upper top surface 138. The circular inner end 164 defines or bounds an annular opening 166 between the cylindrical cavity 162 and the frustum-shaped upper top surface 138 so that the opening 166 is the top of the cylindrical cavity 162. The opening and the lower opening of the frustum-shaped upper top surface 138 are defined.

  Furthermore, the float body 134 is preferably, but not limited to, made of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the float body 134 is constructed from a type of material such as, but not limited to, polystyrene or polycarbonate.

  Outlet passage 170 and extraction passage 180

  7-9, the float body 134 further includes open fluid communication (opened) between the inverted dome-shaped cavity 152 and the lower first through lower volume regions 260 of the float assembly 130. A first circular outlet port 172 disposed in the rounded bottom section 154 of the hemispherical surface 150 inside the float body 134 and a bottom surface 136 of the float body 134 for fluid communication). And an outlet passage 170 extending between the second circular outlet port 174. Further, in one embodiment, the first and second circular outlet ports have a common axis defined by the central longitudinal axis 168 of the float body 134. The second circular outlet port 174 is also surrounded by an annular shoulder 176 that falls into an annular recessed region 178 disposed through the bottom surface 136 of the float body 134. Yes.

  With further reference to FIGS. 7-9, the float body 134 further includes an L-shaped extraction passage 180 defined between the first branch 182, the second branch 184, and the branch bend 186. ing. The first bifurcation 182 is formed by providing a hole in both the outer circumferential surface 142 and the float sidewall 140 at an angle substantially perpendicular to the central longitudinal axis 168 of the funnel float body 134. Surface 142 is then plugged with plug 145. The second branch portion 184 is formed by providing holes in both the frustum-shaped upper top surface 138 and the float side wall 140, and ends at the first branch portion 182 at the bent portion 186, and the funnel float body 134. At an angle substantially parallel to the central longitudinal axis 168. An L-shaped extraction passageway 180 extends and is opened between a teardrop-shaped port 188 disposed in the hemispherical surface 150 and a circular port 190 disposed in the frustum-shaped upper top surface 138. Fluid communication (fluid connection) is performed. The upper portion 192 of the second bifurcation 184 has an increased diameter near the circular port 190 to receive the lower end 128 of the rolled tube 124, so that the inverted dome-shaped cavity 152 and the extraction valve 100 The tube is connected at its upper end 126 to the barb end 122 of the barb joint 114 to provide open fluid communication therebetween (open fluid communication).

  Float cap 194

  Referring to FIGS. 10-13, the funnel and isolation float 132 further includes a float cap 194 that has a circular bottom surface 196, a frustum-shaped lower top surface 198, and a circular bottom surface 196 and a frustum. It has a float cap circumferential side wall 200 extending between the trapezoidal lower top surface 198.

  The circumferential side wall 200 includes an outer circumferential or cylindrical surface 202 that extends between a circumferential bottom surface 196 and the circular outer edge 206 of the frustum-shaped lower top surface 198.

  In order to define an inverted dome-shaped isolation chamber 208 as illustrated in FIG. 6 and close the circular opening 158 of the inverted dome-shaped cavity 152, the float cap 194 is placed inside the cylindrical cavity 162 of the float body 134. Complementary shape and size to fit or fit.

  6 and 10, the frustum-shaped lower top surface 198 of the float cap 194 is a frustum of the float body 134 to define a conical or funnel-shaped surface 210 of the funnel and isolation float 132. It provides continuity below the upper top surface 138 of the shape and defines a conical or funnel shaped cavity 212. In one embodiment, the frustum-shaped upper top surface 138 and the lower top surface 198 are continuous with each other so that the funnel-shaped surface 210 is conical from the circular outer edge 146 of the funnel-shaped upper top surface 138. Even the circular edge 214 on the lower side of the trapezoidal lower top surface 198 is uniformly tapered (tapered). In one embodiment, the lower top surface 198 may be steeper.

  The funnel-shaped surface 210 tapers inwardly from the circular outer edge 146 of the frustum-shaped upper top surface 138 to the lower circular edge 214 of the frustum-shaped lower top surface 198, and the funnel-shaped surface 210. Has moved into the funnel tube portion 216 of the float cap 194. The funnel tube portion 216 defines a cylindrical opening 218 that is open (opened) at a central end that extends through the central region of the float cap 194. Also, at least one funnel fluid passage 220 has a funnel port 222 (FIG. 11) disposed in the lower top surface 198 of the frustum shape and an inlet port 224 (FIG. 12) disposed in the bottom surface 196 of the float cap. Through the funnel float cap 194 and adjacent to the funnel-shaped cavity 212, thereby providing open communication between the funnel-shaped cavity 212 and the inverted dome-shaped isolation chamber 208 ( Open communication: the communication path is open). In one embodiment, there are four funnel fluid passages 220 that are connected between their respective ports 222, 224 and are equally spaced at 90 ° intervals around the funnel tube portion 216. .

  Further, in one embodiment, as illustrated in FIG. 11, the funnel fluid passage 220 is generally triangular in shape and the recessed apex is rounded disposed adjacent to the funnel tube portion 216. And a recessed base that is disposed distally from the funnel tube portion 216 and has a rounded end. Further, in one embodiment, the funnel-shaped surface 210 preferably has a downward slope of about 30 ° from a plane perpendicular to the central axis 226 (FIG. 13) of the float cap 194. Thus, this causes the funnel surface to have an opening of about 120 °.

  12 and 13, one embodiment of the circular bottom 196 of the float cap 194 surrounds a cylindrical opening 218 and a plurality of funnel fluid passages 220 with a central end open (open). An annular portion 230 transitioning from the substantially flat surface 228 and having a downward slope of about 5 ° from a plane perpendicular to the central axis 226 of the funnel cap 134 and a substantially flat annular shape transitioning from the annular portion 230 It has a bottom end face 232.

  The float cap 194 is preferably, but not limited to, a material that is biocompatible and stable to gamma irradiation. In one embodiment, the float cap 194 is composed of, but not limited to, polystyrenes.

  Umbrella valve 234

  With reference to FIGS. 6 and 14, in this one embodiment, the float assembly 130 further includes a first one-way valve or check valve in the form of a resilient umbrella valve 234. The valve is used to selectively open and close the funnel fluid passage 220 based on the body fluid pressure differential across the umbrella valve 234, thereby allowing fluid flow from the funnel-shaped cavity 212 to the inverted dome-shaped isolation chamber 208. While controlling the flow, it prevents the backflow of fluid (body fluid) through the umbrella valve 234 from the reverse dome-shaped isolation chamber 208 to the funnel-shaped cavity 212. Therefore, the umbrella valve 234 is moved from the second or upper volume region 262 above the float to the third volume region or the third volume region defined by the reverse dome-shaped isolation chamber 208 according to the pressure difference of the body fluid applied to the umbrella valve 234. Provides a unidirectional flow of bodily fluid to the isolation area.

  More particularly, in one embodiment, umbrella valve 234 is comprised of a generally circular canopy or dome 236 that is generally perpendicular to a centrally located stem 238 in an unstressed position. Extend to. Stem 238 includes a spherical portion 240 with a tapered shaft portion 242 above the spherical portion. In one embodiment, the umbrella valve 234 is entirely single and of an integral configuration.

  Referring to FIGS. 6, 10, 12, and 14, umbrella valve 234 secures stem 238 through a central end open (open) cylindrical opening 218 disposed through float cap 194. Thus, the float cap 194 is attached. This is achieved by making the length of the stem 238 between the canopy 236 and the spherical portion 240 larger than the length of the cylindrical opening 218 and making the diameter of the spherical portion larger than the diameter of the cylindrical opening 218. As a result, the tapered shaft portion 242 of the stem 238 is inserted into the cylindrical opening 218 from the bottom surface 196 of the float cap 194 and pulled away from the lower top surface 198 of the frustum shape of the float cap 194. When stretched, the elastic bulb 240 passes through a cylindrical opening 218 with an open central end and returns to its normal shape adjacent to the lower top surface 198 of the frustoconical shape of the float cap 194. Thus, it functions as an anchor (fixing tool) for holding the umbrella valve 234 in a fixed position. After the umbrella valve 234 is fixed in place, the tapered shaft portion 242 may be removed while holding the spherical portion 240.

  The generally circular canopy or dome 236 also includes a flat lower contact surface 244 in an unstressed position. Therefore, when the stem 238 located at the center is extended, an axial force is applied to the canopy or dome 236, and as a result, the lower contact surface 244 is extended to a normal tight (close state), and the funnel fluid passage To seal 220, contact sealingly against the substantially flat surface 228 of the bottom surface 196 of the float cap 194.

  In this configuration, the elastic umbrella valve 234 controls fluid (body fluid) flowing into the reverse dome-shaped isolation chamber 208 and prevents fluid (body fluid) flowing out of the reverse dome-shaped isolation chamber 208 via the umbrella valve 234. Provide a one-way valve. In particular, the elastic umbrella valve 234 opens under a predetermined pressure difference or cracking pressure. In other words, the pressure in the funnel fluid passage 220 is caused by the predetermined or known cracking pressure of the umbrella valve 234 to cause the outer surface 246 of the canopy or dome 236. Opens when above or below this side. Thus, if there is a positive pressure difference defined by the pressure in the funnel fluid passage 220 and greater than the pressure on the outer surface 246 of the canopy or dome 236 by an amount greater than the cracking pressure, this pressure difference causes the flexible canopy. Or the dome 236 is flipped or bent downward away from the bottom surface 196 of the float cap 194, thereby allowing body fluid to pass into the reverse dome-shaped isolation chamber 208. After the pressure differential reaches a point or point below the cracking pressure or opening pressure, the flexible canopy or dome 236 recovers its sealing position to a position below the funnel fluid passage 220, thereby creating an inverted dome shape. Fluid from the isolation chamber 208 is prevented from entering further.

  In one embodiment, umbrella valve 234 is preferably, but not limited to, composed of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the umbrella valve 234 is composed of, but not limited to, a silicon-type material, but is compatible with the body fluid in which the material is processed, and preferably biocompatible, and Any flexible and flexible material may be used as long as the material is stable to gamma irradiation. As an example of the umbrella valve 234, Vernay part number 2510-102 is commercially available.

  Duck Bill Valve 248

  With further reference to FIGS. 6, 7 and 15, in one embodiment, the float assembly 130 further comprises a second one-way or check valve in the form of a resilient duckbill valve 248; The duckbill valve selectively opens and closes the outlet passage 170 extending between the first circular outlet port 172 and the second circular outlet port 174 based on the pressure difference of the body fluid applied to the duckbill valve 248. This controls the flow of fluid (body fluid) from the inverted dome-shaped isolation chamber 208 to the first to lower volume region 260 while the first to lower side via the duckbill valve 248. Prevents backflow of fluid (body fluid) from the volume region 260 into the reverse dome-shaped isolation chamber 208. Thus, the duckbill valve 248 provides a unidirectional flow of body fluid from the isolated region in the float to the first to lower volume region 260 in response to the pressure difference of the body fluid applied to the duckbill valve 248.

  In one embodiment, the duckbill valve 248 is connected to the outlet passage 170 and has an elongated outlet slit 256 defined by a pair of elastic sealing lips 258 from a radially outwardly projecting annular flange 252. It is composed of a hollow cylindrical portion 250 having an open end that transitions to a hollow V shape or a converging portion 254 that terminates at the end. These elastic sealing lips 258 are configured to separate to open the slit 256 for the purpose of flowing fluid in one direction through a fluid passage extending axially through the duckbill valve 248.

  The elastic sealing lip 258 is normally maintained at a position where the slit is closed. Then, the pressure inside the hollow V-shaped or converging portion located above the sealing lip 258 becomes larger than the fluid pressure below the elastic sealing lip 258 by the predetermined cracking pressure of the duckbill valve 248. The elastic sealing lip 258 expands (opens) and the slit 256 opens, thereby allowing fluid to pass through the outlet passage or opening 170 and through the fluid passage extending in the axial direction of the duckbill valve 248. Will be able to flow downward. The exit slit 256 is closed when the fluid pressure on the lower or outer surface of the elastic sealing lip 258 is higher than the fluid pressure on the elastic sealing lip 258 above the cracking pressure of the duckbill valve 248, This prevents backflow of fluid through the duckbill valve. Further, when the fluid flow is stopped or in the reverse direction, the back pressure applied by the fluid on the outer surface of the elastic sealing lip 258 engages the lips so as to seal each other, and the outlet slit 256 is closed. Closed to prevent back flow of fluid.

  In the combined state, the annular flange portion 252 of the duckbill valve is seated within an annular recessed region 178 disposed through the bottom surface 136 of the float body 134 while the hollow cylindrical portion 250 is Fitting to the top of the annular shoulder 176, the outlet passage 170 is thus open for communication with the fluid passage extending in the axial direction of the duckbill valve 248, and between the duckbill valve 248 and the float body 134. In order to maintain a tight seal, a press-fit fit or connection is made between the duckbill valve 248 and the annular shoulder 176 of the float body 134.

  In one embodiment, the duckbill valve 248 is preferably, but not limited to, composed of a material that is biocompatible and stable to gamma irradiation, and is not limited to silicon Consists of similar materials. However, any material can be used that is sufficiently elastic and flexible as long as it is compatible with the body fluid to be treated and is preferably biodegradable and stable to gamma irradiation. . An example of a duckbill valve of this example is commercially available as part number DU 054.001-154.01 from MiniValve International.

  Use and operation (action)

  In use and operation, with reference to the figures, the apparatus 10 initially begins with a bodily fluid sample of many components (eg, peripheral blood, bone marrow aspirate, blood components, umbilical cord blood, apheresis blood product, aspirated fat , Semen, urine, breast milk, ascites, exudate, or cerebral bone marrow fluid), the next step: removing the cap 85 from the male luer lock head 88 of the inlet valve 86; Connecting a conventional needleless syringe 270 or other input device containing a body fluid sample to the male luer lock head 88 of the inlet valve 86; a body fluid sample from the conventional needleless syringe 270 or other dispensing device into the device 10; The step of separating the conventional needleless syringe 270 or other dispensing device from the male luer lock head 88 of the inlet valve 86 (FIG. 16). And performing the step of attaching the cap 85 again. When supplying body fluid to the apparatus 10, air is released from the storage container 48 by the air filter.

  The received bodily fluid sample fills the upper to second volume region 262 of the inverted dome-shaped isolation chamber 208 including the funnel-shaped cavity 212 as illustrated in FIG. Flow through the lower first to lower volume region 260 through the lower gap assembly 130 via a circumferential gap 148 located between the surface 142 and the inner circumferential surface 24 of the tube 12. A body fluid sample is initially prevented from flowing into or passing through the reverse dome-shaped isolation chamber 208 by the normally closed umbrella valve 234 and the normally closed duckbill valve 248. In addition, initially, the float assembly 130 is first lifted from the inner surface 30 of the closed bottom end 14 of the tube 12 while protecting the resilient duckbill valve 248 in the cylindrical receiving chamber 45 of the spacing member 34. To provide and prevent a vacuum seal formed therebetween, the circular upper end 38 of the spacing member 34 is adjacent to the circular bottom surface 136 of the float body 134 of the float assembly 130.

  If a bodily fluid sample is received in the storage container 48 defined by the tube 12 and the tube cap 50, the device 10 operates in one or more predetermined times or periods and at one or more predetermined speeds. The centrifuge device 280 is arranged. In one embodiment, the conventional centrifuge 280 operates at about 3200 RPM for about 12 to 15 minutes when blood is used as the body fluid sample. Of course, for specific bodily fluids to be processed by the apparatus 10, one or more time intervals and one or more velocities are determined experimentally or empirically, and may be varied as appropriate according to bodily fluids. .

  During the initial centrifugation of the apparatus 10, the body fluid in the funnel-shaped cavity 212 gradually applies pressure to the umbrella valve 234, which causes the pressure difference on the umbrella valve 234 to become larger than the cracking pressure of the umbrella valve 234. This causes fluid (body fluid) to flow through the one or more funnel fluid passages 220 and into the inverted dome-shaped isolation chamber 208, thereby applying a second pressure to the duckbill valve 248, thereby The pressure difference at 248 is greater than the cracking pressure at the duckbill valve 248 and fluid begins to flow out of the reverse dome-shaped isolation chamber 208 through the outlet passage 170 first. Initially, bodily fluids flow rapidly through the reverse dome-shaped isolation chamber 208 and the hematocrit value does not increase initially if the sample is blood. In the position where both the umbrella valve 234 and the duckbill valve 248 are open, the increase in the body fluid column in the first to lower volume region 260, ie, below the float assembly 130, continues. As this bodily fluid column increases, the device 10 provides a unique circulatory or density (concentration) feedback process, which will be described in detail using peripheral blood as an example of bodily fluids to be processed.

  Therefore, a body fluid such as peripheral blood is stratified based on the density (concentration) of each component that makes up the column, and the cells immediately below the bottom surface 136 of the float body 134 continue to be stratified as well. Thus, the cell density above the level of the duckbill valve 248 is smaller than the cell density below the level of the duckbill valve 248 by an amount that provides a pressure difference less than the cracking pressure of the duckbill valve 248, thereby 248 comes to close. This results in red blood cells accumulating in the bottom section 154 of the reverse dome-shaped isolation chamber 208 over time and clumping, and cells accumulating in the bottom section 154 of the chamber, just below the level of the duckbill valve 248. It has a greater density than cells. This is because the density of red blood cells (red cells) increases in the bottom section 154 and at the same time higher density cells move in the same way toward the bottom end 14 of the tube 12 so the density just below the duckbill valve 248. This is because of the decrease. Thus, as this density increases, the pressure differential will exceed the cracking pressure of the duckbill valve 248 so that the valve will then open, thereby allowing red blood cells to flow through the outlet 170, the first to lower region. 260 replaces bodily fluids, thereby pushing fluid upward through a circumferential gap 148 between the outer circumferential surface 142 of the float sidewall 140 and the inner circumferential surface 24 of the tube 12; A sudden rise in fluid occurs. This upward fluid flow has the effect of returning lighter components of the bodily fluid including white blood cells and platelets from the first to lower volume region 260 of the tube 12 to the upper volume region 262 of the tube 12. Yes. These lighter components are heavier than the plasma in the second to upper volume region 262 in which they circulate, so that the white blood cells and platelets or buffy coat contained therein recede down through the funnel-shaped cavity 212. , Passes through the umbrella valve 234 and is collected in an inverted dome-shaped isolation chamber 208. This cell circulation process or density feedback process continues until equilibrium is reached, in which the pressure difference or density difference between the upper and lower cells of the duckbill valve 248 becomes smaller than the cracking pressure of the duckbill valve 248, and the duckbill valve 248 eventually closes. Similarly, the density difference between the upper and lower cells of the umbrella valve 234 becomes smaller than the cracking pressure of the umbrella valve 234, so that the umbrella valve 234 is finally closed. The process or density feedback process ends.

  Thus, this cell circulation process or density feedback process provides many opportunities to secure white blood cells or other target components of interest within the inverted dome-shaped isolation chamber 208, thereby providing a well-known prior art device and A higher recovery rate can be provided.

  Following the centrifugation step, the device 10 containing the separated and separated body fluid component is removed from the centrifuge device 280 (FIG. 18) and placed on a conventional magnetic stirrer 290 as schematically shown in FIG. Is done. This stirrer is then placed inside an actuated or reverse dome shaped isolation chamber 208 to stir at least one isolated target component inside the isolation chamber 208 for the purpose of increasing recovery in the next collection step. In order to cooperate with the magnetic stir bar 292, a power supply or voltage is applied. In one embodiment, the at least one isolated target component or fraction is agitated for about 20 seconds. As a matter of course, the stirring time may be changed for each body fluid, and the stirring time for a specific body fluid is determined experimentally or empirically.

  Following the agitation step, a step of collecting at least one isolated target component or fraction is performed. This sampling step includes removing the cap 99 from the male luer lock head 102 of the extraction valve 100, connecting a conventional syringe 272 or other extraction device to the male luer lock head 102 of the extraction valve 100, inside At least one isolated target component or fraction from the inverted dome-shaped isolation chamber 208 by supplying a vacuum from the syringe 103 to the tear port 188 located in the rounded bottom section 154 of the hemisphere 150 of And an exit L-shaped extraction passageway 180, a rolled tube 124, a barb disposed from the inverted dome-shaped isolation chamber 208 and through the tear port 188 and in the float body 134 Fitting 114, L-shaped housing passage 62, extraction valve 1 Through the interior of 0, inside the final conventional syringe 272 has a step (19) for collecting at least one isolated target component or fraction.

  After the collection step is completed, the conventional syringe 103 is separated from the male luer lock head 102 of the extraction valve 100 in preparation for at least one diagnostic component or therapeutic application or application of at least one target component or fraction. Is done. When collecting at least one target component or fraction, air enters the storage container 48 by the air filter. Further, if the cracking pressure of the umbrella valve is reached during the sampling step, the interior of the reverse dome-shaped isolation chamber 208 is replaced to replace the removed amount or volume of at least one target component or fraction. Plasma flows in.

  Here, if necessary, a counterweight device 282 may be used in the conventional centrifuge 280 to offset the weight of the device 10 containing bodily fluids.

  In addition, the device 10 should be assembled in an environment that minimizes the risk of particulate matter (contamination risk) in the fluid pathway, and when used in clinical (clinical treatment) applications, the device 10 may not be sterilized. It must be sterilized and the flow path should be non-pyrogenic (free of pyrogens).

  Device 310

  With reference to FIGS. 22 and 23, and in one embodiment, the device 310 defines a main centrifuge tube or container 312 and a storage container 334 for receiving and containing bodily fluids having many components. A tube cap assembly 46 (also shown in detail in FIGS. 3 to 5) for selectively closing the container 312 and slidably disposed within the container 312 and described in detail below. The storage container 334 includes a first to lower volume region 336 below the float assembly 340, a second to upper volume region 338 above the float assembly 340, and an isolation chamber 378 inside the float assembly 340. A dual density subsurface funnel and isolation flow dividing into an isolation or third volume region as defined by In order to move up and down together with the float assembly 340, the float assembly 340 and the tube cap assembly 46 are functionally (operably) connected to each other by a crawler winding structure or a telescopic structure. And a flexible tube 124 (see FIG. 1 for details).

  Main centrifuge tube 312

  More particularly, and with reference to FIG. 24, device 310 includes a closed substantially flat annular bottom end 314, a substantially flat annular top end 316 defining an open top end 318, and a closed bottom. It has a main centrifuge tube or container 312 composed of a cylindrical side wall 320 extending between an end 314 and an annular upper end 316. The cylindrical side wall 320 includes an outer cylindrical surface 322 and an inner circumferential or cylindrical surface 324 that defines a cylindrical storage chamber 326 extending along the central longitudinal axis 328 of the tube 312. This tube is also present at the central longitudinal axis of the device 310.

  In one embodiment, the closed bottom end 314 is formed as a substantially disk-shaped or disk-shaped member having an inner surface 330 and an outer surface 332. The inner surface 330 includes an inner rounded end that transitions into the inner cylindrical surface 324 of the cylindrical side wall 320. Similarly, the outer surface 332 includes an outer rounded end that transitions into the outer cylindrical surface 322 of the cylindrical sidewall 320.

  Further, the closed bottom end 314 may be selectively formed integrally with the cylindrical side wall 320 or configured to cover the side wall 320.

  Furthermore, the tube is preferably, but not limited to, made of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the tube 312 is composed of a material such as, but not limited to, polycarbonate or polystyrene.

  Tube cap assembly 46

  With reference to FIGS. 22 and 23 and returning to FIGS. 3-5, the device 310 further includes a tube cap assembly 46, which has been described in detail above and will not be described for convenience.

  Float assembly 340

  With reference to FIGS. 25 and 26 and as described above, the apparatus 310 further comprises a float assembly comprising a float body 344, a funnel float cap 420, and a bottom float cap 480-500 and an isolating float 42. 340 is further included. Because the funnel and isolation float 342 is precisely made, the funnel and isolation float 342 is a subcomponent consisting of a float body 344, funnel float cap 420, and bottom float caps 480-500 to facilitate accurate injection molding. Preferably, these subcomponents are combined using sonic welding (ultrasonic welding) or other reliable or reliable means.

  The float assembly 340 is in the form of a first check valve means, which in one embodiment is in the form of an umbrella valve 234 detailed in FIG. 14, and in one embodiment is in the form of a duckbill valve 248 detailed in FIG. Second check valve means.

  Float body 342

  More particularly, and with reference to FIGS. 25-27, float body 344, annular bottom end 346, frustum-shaped top surface 348, and annular bottom end 346 and frustum-shaped top top surface 348 And a float body side wall 350 extending therebetween.

The float body side wall 350 includes an outer circumferential or cylindrical surface 352 extending between the annular bottom end 346 and the circular outer edge 354 of the frustum-shaped top surface 348.
The outer circumferential surface 352 of the float body side wall 350 is primarily used to define a circumferential gap (gap) 356 between the outer circumferential surface 352 of the float body side wall 350 and the inner circumference 324 of the tube 312. A diameter smaller than the diameter defined by the inner circumferential surface 324 of the centrifuge tube 312 is defined.

  In addition, the float body side wall 350 includes an inner hemispherical surface 358 that defines an inverted dome-shaped or hemispherical cavity 152 that extends from the rounded bottom section 362 of the hemispherical surface 358 into an inverted dome-shaped configuration. Extending to an upper annular shelf 364 defining an annular opening 366 in the cavity 360 of the first cavity 360. The upper annular shelf 364 transitions into an inner cylindrical surface 368 that defines a cylindrical cavity 370 having an open end that surrounds the inverted dome-shaped cavity 360 so that the annular opening 366 is similarly cylindrical. A lower opening of the cavity 370 is defined. Similarly, the inner cylindrical surface 368 extends from the annular shelf 364 to the circular inner end 372 of the frustum-shaped upper top surface 348. The circular inner end 372 defines an annular opening 374 between the cylindrical cavity 370 and the frustum-shaped upper top surface 348 so that the opening 374 is the top opening and the cone of the cylindrical cavity 370. A lower opening of a trapezoidal top top surface 348 is defined.

  The float body 344 is preferably, but not limited to, made of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the float body 344 is composed of a type of material such as, but not limited to, polystyrene or polycarbonate.

  Exit opening 380

  Referring to FIGS. 27 and 28, the float body 344 further includes open fluid communication (opened) between the inverted dome-shaped cavity 360 and the lower first through lower volume regions 336 of the float assembly 340. A first circular outlet port 382 disposed in the rounded bottom section 362 of the hemispherical surface 358 inside the float body 344 and a bottom surface 386 of the float body 344 for fluid communication). And an outlet opening or passage 380 extending between the second circular outlet port 384.

  Further, the first and second circular outlet ports 382, 384 have a common axis defined by the central longitudinal axis 388 of the float body 344.

  The second circular outlet port 384 is also located within the annular bottom edge 346 of the float body side wall 350 at a position recessed from the annular bottom edge 346 of the float body side wall 350, and this position is four spaced apart. It defines the intersection of two diametrically extending and intersecting cross members (cross members) 390, 392 that define cavities 394, 396, 398, and 400. Four spaced cavities 394, 396, 398, and 400 show the lower outer surface 402 of the dome-shaped cavity 360.

  Extraction opening 404

  Referring to FIGS. 27 and 29, the float body 344 further includes an extraction opening 404, which in one embodiment includes a first branch 406, a second branch 408, and these This is an L-shaped extraction passage 404 defined by a branched bent portion 410 therebetween. The first branch 406 is formed by providing a hole through the float body sidewall 350 at an angle substantially perpendicular to the central longitudinal axis 388 of the float body 344 and adjacent to the outer circumferential surface 352. The portion of the sidewall 350 is then plugged with a plug 412 as illustrated in FIG. The second branch portion 408 is formed by providing holes in both the frustum-shaped upper top surface 348 and the float side wall 350, ends at the first branch portion 406 at the bent portion 410, and An angle substantially parallel to the central longitudinal axis 388. An extraction opening or L-shaped extraction passage 404 extends between an elliptical port 414 disposed in the inner hemisphere 358 and a circular port 416 disposed in the frustum-shaped upper top surface 348. In addition, open fluid communication (fluid connection) is performed. The upper portion 418 of the second bifurcation 408 has an increased diameter near the circular port 416 to receive the lower end 128 of the rolled tube 124, so that the inverted dome-shaped cavity 360 and the extraction valve 100 The tube is connected at its upper end 126 to the barb end 122 of the barb joint 114 to provide open fluid communication therebetween.

  Funnel float cap 420

  Referring to FIGS. 30-33, the funnel and isolation float 342 further includes a funnel float cap 420 that transitions to a disc-shaped base 470 and is installed in a cylindrical shape. It has a frustum or funnel-shaped wall 422 that transitions to the neck 460 and is installed. The funnel-shaped wall 422 includes a conical or funnel-shaped upper surface 424 that defines a truncated cone or funnel-shaped cavity 426. The funnel shaped wall 422 further includes a frustoconical or funnel shaped lower surface 428 and an outer circumferential or cylindrical rim 430 extending between the frustoconical or funnel shaped upper side 424 and the lower side 428. Yes.

  The disk-shaped base 470 is a supplemental shape and size that fits or fits within the cylindrical cavity 370 of the float body 344, and also provides a seal between them when fitted inside. , And a peripheral chamfer 474 on the underside of the disk-shaped base 470 to close the annular opening 366 of the reverse-dome-shaped cavity 360 for purposes of defining the reverse-dome-shaped isolation chamber 378 as illustrated in FIG. And an annular O-ring 472 is captured between the annular shelf 364. Furthermore, the funnel-shaped lower surface 428 of the funnel-shaped wall 422 is complementary to be adjacent to the frusto-conical top surface 348 of the float outer 344 when the annular shelf 364 is sealed by the elastic O-ring 472. Shape and dimensions.

  Means for alignment and connection between the funnel float cap 420 and the float body 344 if the disc-shaped base 470 is properly fitted or fitted into the cylindrical cavity 370 of the float body 344 as described above. Is provided. That is, with reference to FIGS. 25, 27, and 31, the circular segment 432 protruding from the frustum-shaped top surface 348 of the float body 344 has a circular segment notch 434 disposed through the funnel-shaped lower surface 428. Receiving the interior and entering the inside of the funnel-shaped wall 422, the protruding circular segment 432 is adjacent and connected by the circular segment notch 434. Also, the frustum-shaped top surface 348 of the float body 344 includes an annular protrusion 436 that is received within an annular recess 438 (FIG. 25) having a triangular cross-sectional area and a supplemental shape. Dimensions and passes through the funnel-shaped lower side 428 and is disposed within the funnel-shaped wall 422 of the funnel float cap 420. Further, the means for alignment and coupling is similarly aligned or aligned with the opening 440 extending through the funnel-shaped wall 422 with the circular port 416 of the extraction opening or L-shaped extraction passage 404. This causes a portion of the rolled tube 124 adjacent the lower end 128 to be received therethrough.

  30-33, the frustoconical or funnel-shaped upper side 424 of the funnel-shaped wall 422 tapers inwardly from the annular outer edge 442 to the lower annular edge 444; Here, the upper surface 424 of the funnel shape is transferred into the funnel tube portion 446 of the funnel float cap 420. The funnel tube portion 446 defines a cylindrical opening 448 with an open central end that extends through the central portion of the cylindrical neck 460 and the disk-shaped base 470 of the funnel float cap 420. On the other hand, at least one funnel fluid passage or opening 450 is disposed through the funnel float cap 420 at a location adjacent to the funnel tube portion 446, thereby isolating the funnel-shaped cavity (FIG. 25) and the inverted dome shape. Open communication with the chamber 378 is provided. In one embodiment, there are four funnel fluid passages 450 equally spaced at 90 ° intervals around the funnel tube portion 446.

  Further, in one embodiment, the funnel fluid passage or opening 450 is generally triangular and has a rounded end adjacent to the funnel tube portion 446, as illustrated in FIGS. It has a concave tip and a convex base with a rounded end located away from the funnel tube portion 446. In one embodiment, the funnel-shaped upper surface 210 has a downward slope of about 30 ° from a plane perpendicular to the central axis 452 (FIG. 31) of the float cap 420. As a result, an opening of about 120 ° is provided on the funnel surface.

  Further, and with reference to FIGS. 31 and 33, the lower peripheral chamfer 474 transitions into an annular recessed bottom region 476, while this bottom region is centered as illustrated in FIG. And an open cylindrical opening 448 and a plurality of funnel fluid passages or openings 450 within a substantially flat bottom surface 478.

The funnel float cap 420 is preferably, but not limited to, a material that is biocompatible and stable to gamma irradiation.
In one embodiment, funnel float cap 420 is constructed from a material such as, but not limited to, polystyrene or polycarbonate.

  Bottom cap 480

  34-36, the funnel and isolation float further includes a bottom float cap 480 that has a substantially flat top surface 484, a substantially flat bottom surface 486, and A circular wall 482 having an outer peripheral edge 488 extending therebetween is provided. In addition, the bottom float cap 420 has an annular lip or protrusion 490 that protrudes from a substantially flat top surface 484 at a location adjacent to the outer circumferential edge 488 of the circular wall 482. , And a triangular shaped cross-sectional area sized to be received within a complementary shaped annular recess 538 (FIG. 25) disposed within the float body 344. The bottom float cap 480 also has a centrally located annular shelf 492 that is recessed below the substantially flat top surface 484 of the circular wall 482. The shelf 492 disposed in the center surrounds the centrally located opening 492 disposed through the circular wall 482. In addition, the bottom float cap 480 has a plurality of spaced fins 496 that project downward from the substantially flat bottom surface of the circular wall 482. In one embodiment, there are four downwardly projecting fins 496 spaced equidistantly at 90 ° around an opening 494 located in the center of the bottom float cap 480. In use and operation, these fins 496 protect the float assembly 340 from the closed bottom end of the tube 312 to protect the elastic duckbill valve 248 in the initial state while preventing the vacuum seal formed therebetween. It functions to first lift off the inner surface 330 of 314.

  The bottom float cap 480 is preferably, but not limited to, composed of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the bottom float cap 480 is composed of a material such as, but not limited to, polystyrene or polycarbonate.

  Bottom cap 500

  Referring to FIGS. 37-39, and in another embodiment, the funnel and isolation float 342 further includes a bottom float cap 500 that has a substantially flat top surface 504, substantially flat. And a circular wall 502 comprising an outer peripheral edge 508 extending therebetween. In addition, the bottom float cap 500 has an annular lip protrusion 510 that protrudes from a substantially flat top surface 504 at a location adjacent the outer peripheral edge 508 of the circumferential wall 502, and It has a triangular cross-sectional area dimensioned to be received within a complementary shaped annular recess 538 (FIG. 25) disposed within the float body 344. In addition, the bottom float cap 500 has a centrally located annular shelf 512 that is recessed below the substantially flat top surface 504 of the circular wall 502. A centrally disposed annular shelf 512 surrounds a centrally located opening 514 disposed through the circular wall 502. Further, the bottom float cap 500 has a plurality of spaced cylindrical legs 516 that project downwardly from the substantially flat bottom surface 506 of the circular wall 502. In one embodiment, there are four downwardly projecting cylindrical legs 516 that are spaced equidistantly at 90 ° around an opening 514 located in the center of the bottom float cap 500. Each cylindrical leg 516 extends downwardly from an open end disposed within the substantially flat top surface 504 of the circular wall 502. In use and operation, these cylindrical legs 516 prevent the float assembly 340 from closing the tube 312 to protect the elastic duckbill valve 248 in the initial state while preventing the vacuum seal formed therebetween. It functions to lift first from the inner surface 330 of the bottom end 314. The bottom float cap 500 is preferably, but not limited to, made of a material that is biocompatible and stable to gamma irradiation. In one embodiment, the bottom float cap 500 is composed of a material such as, but not limited to, polystyrene or polycarbonate.

40 and 41, each blind hole 518 is a carrier for cylindrical weights (weights) inserted therein for the purpose of adjusting or controlling the overall density of the float assembly 340. It can also function as a support place. As such, one or more weights 530 can be coupled to the bottom float cap 500 to provide a ballast for adjusting or controlling the overall density of the float assembly 340. A preferred material for the ballast or weight for adjusting the specific gravity of the float assembly 340 is a metal rod such as stainless steel. For application or application of the present invention to blood and bone marrow processing to collect buffy coat, it is preferable to adjust the float assembly 340 between the density of plasma and concentrated red blood cells, more specifically, Preferred assembly densities are in the range of 1.02 to 1.08 grams / cm ³ , most preferably in the range of 1.03 to 1.07 grams / cm ³ . The specific gravity used may be optimized for the desired therapeutic and diagnostic application with respect to the cellular composition produced by the device. The higher the specific gravity, the greater the use of red blood cells, and the higher the efficiency of white blood cell (WBC) recovery.

  Referring to FIGS. 42 and 43, each cylindrical weight 530 may be supported by each gantry structure 532, which is formed on the flat top surface 484 of the bottom float cap 480 and has a bottom portion. When the float cap 480 mates with the float body 344, it is received within one of the four spaced cavities 394, 396, 398, and 400 in the bottom surface 386 of the float body 344 (FIG. 28). Accordingly, one or more weights 530 are coupled to the bottom float cap 480 to provide ballast for adjusting or controlling the overall density of the float assembly 340 as described in detail above.

In addition, and with reference to FIG. 44, a cylindrical weight 300 may be received within the blind hole 302 to fine tune or control the overall density of the float assembly 340. As such, one or more weights 300 can be coupled to the float body 134 to provide a ballast for adjusting or controlling the overall density of the float assembly 130. In this embodiment, the preferred material for the ballast or weight for adjusting the specific gravity of the float assembly 130 is also a metal rod, such as stainless steel. In addition, for application or application of the present invention to blood and bone marrow processing for collecting buffy coat, it is preferable to adjust the float assembly 130 between the density of plasma and concentrated red blood cells, more specifically. The preferred assembly density is in the range of 1.02 to 1.08 grams / cm ³ and most preferably in the range of 1.03 to 1.07 grams / cm ³ . The specific gravity used may be optimized for the desired therapeutic and diagnostic application with respect to the cellular composition produced by the device. The higher the specific gravity, the greater the use of red blood cells, and the higher the efficiency of white blood cell (WBC) recovery.

  Umbrella valve 234

  Referring to FIGS. 25, 30 to 33 and again to FIG. 14, in this embodiment, the float assembly 340 further includes a first one-way valve or check valve in the form of a resilient umbrella valve 234. This one-way valve is used to selectively open and close the funnel fluid passage 450 based on the body fluid pressure differential across the umbrella valve 234, thereby allowing the funnel-shaped cavity 426 to be inverted dome-shaped. Controls the flow of fluid to the isolation chamber 378 while preventing backflow of fluid (body fluid) through the umbrella valve 234 from the inverted dome-shaped isolation chamber 378 to the funnel-shaped cavity 426. Accordingly, the umbrella valve 234 is configured to change from the second or upper volume region 338 above the float to the third volume region defined by the reverse dome-shaped isolation chamber 378 in accordance with the pressure difference of the body fluid applied to the umbrella valve 234. Provides a unidirectional flow of bodily fluid to the isolation area.

  More particularly, and as illustrated above and illustrated in FIG. 14, in one embodiment, umbrella valve 234 is comprised of a generally circular canopy or dome 236 that is centrally located in an unstressed position. It extends substantially perpendicular to the stem (bar) 238 located at. Stem 238 includes a spherical portion 240 with a tapered shaft portion 242 above the spherical portion. In one embodiment, the umbrella valve 234 is entirely single and of an integral configuration.

  Referring to FIGS. 25, 30, 33 and again to FIG. 14, umbrella valve 234 secures stem 238 through a centrally open cylindrical opening 448 disposed through funnel float cap 420. Is attached to the funnel float cap 420. This is achieved by making the length of the stem 238 between the canopy 236 and the spherical portion 240 larger than the length of the cylindrical opening 448 and making the diameter of the spherical portion larger than the diameter of the cylindrical opening 448. As a result, the tapered shaft portion 242 of the stem 238 is inserted into the cylindrical opening 448 from the substantially flat bottom surface 478 of the funnel float cap 420 and away from the funnel-shaped upper side 424 of the funnel float cap 420. When pulled and stretched, the elastic bulb 240 passes through a cylindrical opening 448 having an open central end and is adjacent to the funnel-shaped upper side 424 of the funnel float cap 420 in its normal shape. Thus, it functions as an anchor (fixing tool) for holding the umbrella valve 234 in a fixed position. After the umbrella valve 234 is fixed in place, the tapered shaft portion 242 may be removed while holding the spherical portion 240.

  The generally circular canopy or dome 236 also includes a flat lower contact surface 244 in an unstressed position. Thus, when the central stem 238 is extended, an axial force is applied to the canopy or dome 236, resulting in the lower side 244 being extended to a normal tight (close contact) condition and a funnel fluid passage or opening 450. To seal against the substantially flat bottom surface 478 of the funnel float cap 420.

  In this configuration, the elastic umbrella valve 234 controls the fluid (body fluid) flowing into the reverse dome-shaped isolation chamber 378 and prevents the fluid (body fluid) flowing out of the reverse dome-shaped isolation chamber 378 via the umbrella valve 234. Provide a one-way valve. In particular, the elastic umbrella valve 234 opens under a predetermined pressure difference, that is, cracking pressure. In other words, the pressure in the funnel fluid passage or the opening 450 is reduced by the predetermined or known cracking pressure of the umbrella valve 234, so that the outer surface of the canopy or dome 236. 246 is opened when it is above or below this 246. Thus, if there is a positive pressure difference defined by the pressure in the funnel fluid passage or opening 450 and greater than the pressure on the outer surface 246 of the canopy or dome 236 by an amount greater than the cracking pressure, this pressure difference causes the flexibility. One canopy or dome 236 flips or bends downward away from the substantially flat bottom surface 478 of the funnel float cap 420, thereby allowing body fluid to pass into the reverse dome-shaped isolation chamber 378. After the pressure differential reaches a point or point below the cracking pressure or opening pressure, the flexible canopy or dome 236 recovers its sealing position to a position below the funnel fluid passage or opening 450, thereby reversing. Further entry of body fluid from the dome-shaped isolation chamber 378 is prevented.

  Duck Bill Valve 248

  25, 27, 34-39 and again with reference to FIG. 15, one embodiment of the float assembly 340 further comprises a second one-way valve or check valve in the form of a resilient duckbill valve 248. The one-way valve has an outlet passage or opening 380 extending between the first circular outlet port 382 and the second circular outlet port 384 based on the body fluid pressure difference across the duckbill valve 248. Used to selectively open and close, thereby controlling the flow of fluid from the inverted dome-shaped isolation chamber 378 to the first to lower volume region 336, while from the first to lower volume region 336. Prevents backflow of fluid (body fluid) through the duckbill valve 248 into the reverse dome-shaped isolation chamber 378. Thus, the duckbill valve 248 provides a unidirectional flow of body fluid from the isolation chamber 378 within the float assembly 340 to the first to lower volume region 336 in response to the pressure differential of the body fluid applied to the duckbill valve 248. .

  In one embodiment, and as illustrated above and illustrated in FIG. 15, the duckbill valve 248 is coupled to the outlet passage 170 and from a radially outwardly projecting annular flange portion 252 for a pair of elastic sealing. It is composed of a hollow cylindrical portion 250 with an open end that transitions to a hollow V-shaped or converging portion 254 that terminates in an elongated exit slit 256 defined by a lip 258. These elastic sealing lips 258 are formed away to open the slit 256 for the purpose of flowing fluid in one direction through a fluid passage extending axially through the duckbill valve 248.

  The elastic sealing lip 258 is normally maintained at a position where the slit is closed. Then, the pressure inside the hollow V-shaped or converging portion located above the sealing lip 258 becomes larger than the fluid pressure below the elastic sealing lip 258 by the predetermined cracking pressure of the duckbill valve 248. The elastic sealing lip 258 expands (opens) and the slit 256 opens, thereby allowing fluid to pass through the outlet passage or opening 170 and through the fluid passage extending in the axial direction of the duckbill valve 248. Will be able to flow downward. The exit slit 256 is closed when the fluid pressure on the lower or outer surface of the elastic sealing lip 258 is higher than the fluid pressure on the elastic sealing lip 258 above the cracking pressure of the duckbill valve 248, This prevents backflow of fluid through the duckbill valve. Further, when the fluid flow is stopped or in the reverse direction, the back pressure applied by the fluid on the outer surface of the elastic sealing lip 258 engages the lips so as to seal each other, and the outlet slit 256 is closed. Closed to prevent back flow of fluid.

  In the combined condition, the annular flange portion 252 of the duckbill valve sits between the bottom surface 386 of the float body 344 and the annular shelf portion 492 of the float cap 480 or the annular shelf portion 512 of the bottom float cap 500, The cylindrical portion 250 passes through a centrally located opening 494 or 514, respectively, so that a hollow V-shaped or converging portion 254 extends under the bottom float cap 480 or 500, respectively. In this configuration, the outlet passage or opening 380 is in open communication with the axially extending fluid passage of the duckbill valve 248, and the bottom surface 386 of the float body 344, the duckbill valve 248, and the duckbill valve 248 and the float body 344. In order to maintain a hermetic seal, a press fit fit or connection is made between the annular shelf 492 of the bottom float cap 480 or the annular shelf 512 of the bottom float cap 500.

  Use and operation (action)

  In use and operation, with reference to the figures, device 310 conforms to the use and operation described above for device 10. The device 310 initially receives a body fluid sample of many components (eg, peripheral blood, bone marrow aspirate, blood components, umbilical cord blood, apheresis blood product, semen, urine, breast milk, ascites, exudate, or cerebral bone marrow fluid). In order to achieve this, the following steps are taken: a step of removing the cap (lid) 85 from the male luer lock head 88 of the inlet valve 86; a conventional fluid sample contained in the male luer lock head 88 of the inlet valve 86; Connecting a needleless syringe 270 or other dispensing device, injecting or dispensing a body fluid sample into the device 310 from the conventional needleless syringe 270 or other dispensing device, a conventional needleless syringe 270 or other dispensing device Separating the male luer lock head 88 of the inlet valve 86 and reattaching the cap 85. When supplying body fluid to the apparatus 310, air is released from the storage container 334 by the air filter.

  The received bodily fluid sample fills the upper to second volume region 338 of the inverted dome-shaped isolation chamber 378 including the funnel-shaped cavity 426, and the outer circumferential surface 352 of the float sidewall 350 and the inner circle of the tube 312. It flows to a lower first through lower capacity region 336 through a circumferential gap 356 disposed between the peripheral surface 324 and the float assembly 340. A body fluid sample is initially prevented from flowing into or passing through the inverted dome-shaped isolation chamber 378 by the normally closed umbrella valve 234 and the normally closed duckbill valve 248. In addition, the spaced fins 496 of the bottom float cap 480 or the spaced cylindrical legs 516 of the bottom float cap 500 protect the resilient duckbill valve 248 in the initial state while the inner surface of the closed bottom end 314 of the tube 312. The initial lift from 330 to the float assembly 340 is provided, preventing the vacuum seal formed therebetween.

  If a bodily fluid sample is received within a storage container 334 defined by a tube 312 and a tube cap 50, the device 310 operates in one or more predetermined times or periods and at one or more predetermined speeds. The centrifuge device 280 is arranged. In one embodiment, the conventional centrifuge 280 operates at about 3200 RPM for about 12 to 15 minutes when blood is used as the body fluid sample. Of course, for particular bodily fluids processed by the device 310, one or more time intervals and one or more velocities are determined experimentally or empirically, and may be varied as appropriate according to bodily fluids. .

  During the initial centrifugation of the device 310, body fluid in the funnel-shaped cavity 426 gradually applies pressure to the umbrella valve 234, which causes the pressure difference on the umbrella valve 234 to become larger than the cracking pressure of the umbrella valve 234. This causes fluid (body fluid) to flow through the one or more funnel fluid passages or openings 450 and into the inverted dome-shaped isolation chamber 378. Therefore, a second pressure is applied to the duckbill valve 248, whereby the pressure difference of the duckbill valve 248 becomes larger than the cracking pressure of the duckbill valve 248, and the body fluid is reverse dome-shaped through the outlet passage or opening 380. From the isolation chamber 378 first. Initially, bodily fluids flow quickly through the reverse dome-shaped isolation chamber 378 and the hematocrit value does not increase initially if the sample is blood. In the position where both the umbrella valve 234 and the duckbill valve 248 are open, the column of body fluid in the first to lower volume region 336, ie, below the float assembly 340, continues. As this bodily fluid column grows, the device 310 provides a unique circulatory or density (concentration) feedback process, which is described in detail using peripheral blood as an example of bodily fluids to be processed.

  Accordingly, a body fluid such as peripheral blood is stratified based on the density (concentration) of each component making up the column, and the cells immediately below the bottom surface 484 of the bottom float cap 480 or the bottom surface 506 of the bottom float cap 500 are Continue stratification as well, so that the cell density above the level of the duckbill valve 248 provides a pressure difference that is less than the cracking pressure of the duckbill valve 248 than the cell density below the level of the duckbill valve 248. The duckbill valve 248 is closed by this amount. As a result, red blood cells accumulate over time in the bottom section 362 of the inverted dome-shaped isolation chamber 378 and clump, and cells accumulate in the bottom section 362 of the chamber, just below the level of the duckbill valve 248. It has a greater density than cells. This is because the density of red blood cells (red cells) increases in the bottom section 362, while at the same time larger density cells move in the same way toward the bottom end 314 of the tube 312 so the density just below the duckbill valve 248. This is because of the decrease. Thus, as this density increases, the pressure differential will exceed the cracking pressure of the duckbill valve 248 so that the valve is then opened, thereby allowing red blood cells to flow through the outlet passage or opening 380, the first to lower Fluid in the side region 336, thereby pushing fluid upward through a circumferential gap 356 between the outer circumferential surface 352 of the float sidewall 350 and the inner circumferential surface 324 of the tube 312. Or a sudden rise in fluid occurs. This upward fluid flow has the effect of returning lighter components of the body fluid, including white blood cells and platelets, from the first to lower volume region 336 of tube 312 to the upper volume region 338 of tube 312. These lighter components are heavier than the plasma in the second to upper volume region 338 they circulate, so the white blood cells and platelets or buffy coat contained therein recede down through the funnel-shaped cavity 426. Then passes through the umbrella valve 234 and is collected in an inverted dome-shaped isolation chamber 378. This cell circulation process or density feedback process continues until equilibrium is reached, in which the pressure difference or density difference between the upper and lower cells of the duckbill valve 248 becomes smaller than the cracking pressure of the duckbill valve 248, and the duckbill valve 248 eventually closes. Similarly, the difference in density between the upper and lower cells of the umbrella valve 234 becomes smaller than the cracking pressure of the umbrella valve 234 so that the umbrella valve 234 eventually closes, thereby causing the characteristic cell circulation of the device 310. The process or density feedback process ends.

  Thus, this cell circulation process or density feedback process provides many opportunities to secure white blood cells or other target components of interest within the inverted dome-shaped isolation chamber 378, thereby providing well-known prior art devices and A higher recovery rate can be provided.

  Following the centrifugation step, the device 310 containing the separated and separated body fluid components is removed from the centrifuge device 280 and placed on a conventional magnetic stirrer 290 as shown schematically in FIG. This stirrer is then placed inside an actuated or reverse dome shaped isolation chamber 208 to agitate at least one isolated target component inside the isolation chamber 378 for the purpose of increasing recovery in the next collection step. In order to cooperate with the magnetic stir bar 292, a voltage is applied. In one embodiment, the at least one isolated target component or fraction is agitated for about 20 seconds. As a matter of course, the stirring time may be changed for each body fluid, and the stirring time for a specific body fluid is determined experimentally or empirically.

  Following the agitation step, a step of collecting at least one isolated target component or fraction is performed, the step of removing the cap 99 from the male luer lock head 102 of the extraction valve 100, the extraction valve 100 Connecting a conventional syringe 272 or other extraction device to the male luer lock head 102, the extraction passage or opening 404 from the syringe 103, and the rounded bottom section 362 of the inner hemisphere 358. Collecting at least one isolated target component or fraction from the reverse dome-shaped isolation chamber 378 by supplying a vacuum to the closed port 414, and from the reverse dome-shaped isolation chamber 378 and through the port 414. And in the float body 344 Collecting at least one isolated target component or fraction through the extracted extraction passage or opening 404, the L-shaped housing passage 62, the extraction valve 100, and finally into the conventional syringe 272. Have.

  After the collection step is completed, the conventional syringe 103 is separated from the male luer lock head 102 of the extraction valve 100 in preparation for at least one diagnostic component or therapeutic application or application of at least one target component or fraction. Is done. When collecting at least one target component or fraction, air enters the storage container 48 by the air filter. In addition, if the cracking pressure of the umbrella valve is reached during the sampling step, the interior of the reverse dome-shaped isolation chamber 378 is replaced to replace the removed amount or volume of at least one target component or fraction. Plasma flows in.

  Here, a counterweight device 282 may be used with the conventional centrifuge 280 to offset the weight of the device 310 containing bodily fluids as needed.

  In addition, the device 310 should be assembled in an environment to minimize the risk of particulate matter (contamination risk) in the fluid pathway, and when used in clinical (clinical treatment) applications, the device 310 may be sterilized or disinfected. And the flow path should be non-pyrogenic (free of pyrogens).

  In addition, the materials used in the construction of various embodiments of the device for separating and isolating components of body fluids are biocompatible, including plastics, rubber, metals and magnets, and are substantially free of cytotoxic leaching. It is something that is not. The preferred plastic for constructing the tube is polystyrene or polycarbonate. The plastic for the construction of the funnel and isolation float body is polystyrene or polycarbonate. A preferred material for the one-way valve in the float is a silicone rubber that is stable to gamma irradiation. The preferred material for the ballast or weight to adjust the specific gravity of the funnel assembly is a metal rod such as stainless steel, which is not allowed to contact biological components and is located in the interior of the isolation float Placed in. The preferred magnetic stir bar located in the isolation chamber of the funnel and isolation float is made from a magnetic material such as a neodymium magnet. Magnetic stir bars can be used in a variety of shapes and sizes, but are compatible with the specific gravity of the float and provide the potential for magnetic stirrer that obstructs or blocks the flow of fluid within the outlet opening of the float inner chamber. Should be avoidable. A single magnetic rod having a dimension about half the inner diameter of the inside of the isolation chamber is suitable.

  A preferred method for preparing tube examples, tube caps, funnels and isolating float examples is injection molding. Alternatively, the funnel and isolation float can be manufactured using tools or tools in machine shops such as lathes and drills. The rolled tube used to connect the lid to the float is preferably medical grade polyvinyl chloride. Since the float needs to be made precisely or precisely, it is preferable to make the device as a sub-component to facilitate precision injection molding, and these sub-components are sonic welded (ultrasonic welding) or other reliable or reliable It is preferable to combine using some means.

  The above-described apparatus embodiments 10 and 310 including the method of use and operation are merely examples of industrial applications of the present invention.

  Accordingly, various other structural changes and applications are obvious without departing from the scope and technical idea of the present invention described in the claims.

DESCRIPTION OF SYMBOLS 10 Apparatus 12 Centrifugation tube 26 Storage chamber 34 Separation member 46 Tube cap assembly 48 Storage container 80 Air filter 86 Inlet valve 100 Extraction valve 124 Tube 130 Float assembly 132 Funnel and isolation float 134 Float body 194 Float cap 208 Isolation chamber 310 equipment 340 float assembly 342 funnel and isolation float 344 float body 362 bottom section 420 funnel float cap 480 bottom cap

Claims (44)

A device for separating and isolating the components of bodily fluid by centrifugation, wherein the device contains a bodily fluid having many components;
A float slidably disposed within the storage container and having an internal isolation chamber;
A first valve means for injecting body fluid into the internal isolation chamber of the float in response to a first pressure difference of body fluid on the first valve means; and A second valve means for draining body fluid from the internal isolation chamber of the float in response to a second pressure difference of the body fluid on the second valve means. Become
Under centrifugation of the device, the first valve means and the second valve means are initially configured for the body fluid to isolate at least one target component of body fluid within the internal isolation chamber of the float. Each component is allowed to flow through the internal isolation chamber and then in response to the first pressure difference of the body fluid on the first valve means and the body fluid on the second valve means An apparatus that closes and seals the internal isolation chamber in response to a second pressure differential.   The float is in open communication with the internal isolation chamber and external to the storage container to permit extraction of at least one target component of body fluid from the internal isolation chamber of the float to the exterior of the storage container. The apparatus of claim 1, comprising a derived flexible tube.   The storage container comprises a container having a closed bottom end, an open top end, and a circumferential side wall extending between the closed bottom end and the open top end, the circumferential side wall comprising: To have an inner circumferential surface defining a storage chamber extending along a central longitudinal axis of the container, and the storage container defines the storage container for storing bodily fluids having many components The apparatus of claim 2, further comprising a cap for selectively closing the open upper end of the container.   To define the inner isolation chamber as an inverted dome-shaped isolation chamber, the float comprises a lower side having an inverted hemispherical surface with an upper annular shelf transitioning into an upper ceiling surface. Item 3. The apparatus according to Item 3.   The float surrounds the lower side and funnel to facilitate the flow of the body fluid into the internal isolation chamber in response to the first pressure difference of the body fluid on the first valve means. A funnel-shaped cavity defining a funnel-shaped cavity converging toward the inverted dome-shaped chamber and opening with the first valve means. The apparatus of claim 4 in communication.   The apparatus of claim 5, wherein said first valve means comprises an umbrella valve.   The apparatus of claim 6 wherein said second valve means comprises a duckbill valve. The float, the first valve means, and the second valve means have an overall selected density of about 1.02 grams / cm ³ to about 1.08 grams / cm ^3. The device described.   A magnetic stir bar disposed in the internal isolation chamber of the float to agitate at least one target component of bodily fluid in the internal isolation chamber of the float when actuated by magnetism; The apparatus of claim 1. A device for separating and isolating components of body fluid by centrifugation,
A storage container having an inner circumferential surface defining a chamber for storing a body fluid having many components;
It has a float that is slidably arranged inside the storage container and divides the storage container into a lower capacity area and an upper capacity area,
The float has an inner isolation chamber that defines an intermediate volume region and an outer circumferential surface that is spaced circumferentially from the inner circumferential surface of the storage container and defines a circumferential gap therebetween. And
A first valve means for opening and closing a body fluid communication path from the upper volume region to the inner isolation chamber inside the float; and from the inner isolation chamber inside the float to the lower part of the container A second valve means for opening and closing the body fluid communication path to the side volume region;
To isolate at least one target component of a multi-component body fluid within the inner isolation chamber of the float, under centrifugation, in response to a first pressure difference of the body fluid on the first valve means, and In response to a second pressure difference of the body fluid on the second valve means, the lower volume from the upper volume region into the inner isolation chamber and out of the inner isolation chamber into the lower volume region The first valve means and the second valve means control the circulation of at least one closed-loop body fluid leaving the area, through the circumferential gap, back into the upper volume area and back into the inner isolation chamber. Do the equipment.   11. The apparatus of claim 10, wherein the float includes an upper funnel surface for facilitating the flow of bodily fluid into the inner isolation chamber in response to the first pressure differential of bodily fluid on the first valve means. .   The apparatus of claim 11, wherein the inner isolation chamber is formed as an inverted dome shaped chamber.   Means for extracting at least one target component from the reverse dome shaped chamber by aspirating at least one target component from a lower arcuate portion of the reverse dome shaped chamber; The apparatus of claim 12.   14. The apparatus of claim 13, wherein the first valve means is an umbrella valve.   The apparatus of claim 14, wherein said second valve means is a duckbill valve.   16. The apparatus according to claim 15, further comprising lift-off means for initially pulling the float away from the inner bottom surface of the storage container. The float, the first valve means, and the second valve means have an integral, selected density of about 1.02 grams / cm ³ to about 1.08 grams / cm ³ . The apparatus of claim 10.   A magnetic stirring bar disposed in the inner isolation chamber of the float for stirring at least one target component of body fluid within the inner isolation chamber of the float when magnetically driven; The apparatus of claim 10. A device for separating and isolating components of bodily fluids by centrifugation, said device comprising:
A closed bottom end, an open top end, and a container side wall extending between the closed bottom end and the open top end, the side wall extending along a central longitudinal axis of the container A container having an inner circumferential surface defining a containing chamber to be
A cap for selectively closing the open upper end of the container to define a storage container for containing a body fluid having a number of components;
A float assembly that is slidably disposed within the container and divides the container into a lower volume area and an upper volume area;
The float assembly comprises:
A lower cylindrical portion including an inverted hemispherical inner surface surrounded by an inner ceiling surface to define an inverted dome-shaped chamber for isolating a target component of bodily fluid;
The lower cylinder of the float for enclosing, directing, and promoting (facilitating) the flow of bodily fluid from the upper volume region to the inverted dome-shaped chamber while surrounding the lower cylindrical portion An upper cylindrical portion including a conical upper surface defining a funnel-shaped cavity converging towards the portion;
An open-ended inlet passage disposed within the float to provide open fluid communication between the funnel-shaped cavity and the inverted dome-shaped chamber;
An outlet passage having an open end disposed within the float to provide open communication between the inverted dome-shaped chamber and the lower volume region;
First valve means for controlling fluid flow from the funnel-shaped cavity to the inverted dome-shaped chamber and from the inverted dome-shaped chamber through the first valve means to the funnel-shaped cavity; First valve means for selectively opening and closing an inlet passage open to the end in response to a first pressure difference on the first valve means to prevent back flow of fluid to the first valve means;
Second valve means for controlling fluid outflow from the reverse dome shaped chamber to the lower volume area of the container and from the lower volume area through the second valve means to the reverse dome. A second valve means for selectively opening and closing the outlet passage open to the end in response to a second pressure difference on the second valve means to prevent back flow of fluid into the shaped chamber; , And
In order to isolate the target component of bodily fluid inside the inverted dome-shaped chamber of the float during centrifugation, the first valve means and the second valve means are initially configured so that the component of the bodily fluid Allowing flow through the inverted dome-shaped chamber, and then in response to the first pressure differential on the first valve means and the second pressure differential on the second valve means In response, the device is closed to seal the inverted dome shaped chamber.   The flexible tube further comprises a wound flexible tube that provides a vacuum to the extraction valve connected to the cap for the body fluid isolated within the inverted dome-shaped chamber. In order to extract the target component, one end is functionally connected to an extraction valve connected to the cap, and the other end is disposed in the upper cylindrical portion and the lower cylindrical portion of the float assembly. 20. The apparatus of claim 19, wherein the apparatus is operably connected to one end of an open extraction passage and the other end terminates in a bottom region of the inverted dome-shaped inner surface of the lower cylindrical portion of the float assembly. .   20. The apparatus of claim 19, further comprising inlet valve means coupled to the cap and in open communication with the containment chamber for dispensing bodily fluid having a number of components into the containment chamber.   When extracting a target component isolated in the inverted dome-shaped chamber to connect the cap and to release air from the storage chamber when a body fluid having many components is distributed in the storage chamber 20. The apparatus of claim 19, further comprising air filter means in open communication with the containment chamber to allow air to enter the containment chamber.   20. The apparatus of claim 19, further comprising lift-off means for initially pulling the float away from the inner bottom surface of the container and preventing a vacuum seal from being formed therebetween.   The apparatus of claim 19, wherein the first valve means comprises an umbrella valve.   The apparatus of claim 19, wherein said second valve means comprises a duckbill valve. The apparatus of claim 19, wherein the float assembly has a selected density of about 1.02 grams / cm ³ to about 1.08 grams / cm ³ .   In order to stir the target component of the body fluid inside the inverted dome-shaped chamber of the float when magnetically driven, it further has a magnetic stir bar disposed in the inverted dome-shaped chamber of the float. The apparatus of claim 19. A device for separating and isolating components of bodily fluids by centrifugation, said device comprising:
A storage container for storing a body fluid having many components;
A float that is slidably disposed within the storage container and has an inner isolation chamber;
The float has a first opening means, and the first opening means corresponds to the first pressure difference of the body fluid in the body fluid above and below the first opening means. Allow the flow of bodily fluids into the inner isolation chamber,
The float has a second opening means, and the second opening means is configured to change the float of the float according to a second pressure difference of the body fluid in the upper and lower body fluids of the second opening means. A device that allows the flow of bodily fluids out of the inner isolation chamber.   Further comprising third opening means, the third opening means for extracting at least one target component of bodily fluid from the interior of the inner isolation chamber for the third opening means inside the inner isolation chamber. Body fluid to the outside of the inner isolation chamber of the float in response to a third pressure differential of body fluid on the body fluid on the first port and the second port of the third opening means outside the isolation chamber 30. The device of claim 28, wherein the device allows flow.   When the apparatus is centrifuged, the first opening means and the second opening means initially flow in through the first opening means and flow out through the second opening means. Thereby permitting the flow of bodily fluid components through the isolation chamber, and then the first pressure difference of the bodily fluid on the upper and lower body fluids of the first opening means and the upper side of the second opening means and In response to a second pressure difference of the body fluid applied to the lower body fluid, the body fluid flows through the inner isolation chamber by flowing in through the first opening means and flowing out through the second opening means. 30. The apparatus of claim 28, wherein the apparatus reduces component flow.   29. The apparatus of claim 28, wherein the first opening means includes an opening area that is at least twice as large as the opening area of the second opening means.   29. The apparatus of claim 28, wherein the float is freely slidable within the storage container before centrifugation, during centrifugation, and after centrifugation.   30. The device of claim 28, wherein the bodily fluid is derived from a mammal and comprises bone marrow, whole blood, blood components, semen, fat, ascites, cerebral bone marrow fluid, or mixtures thereof. 30. The apparatus of claim 28, wherein the float has a selected density of about 1.02 grams / cm < ³ > to about 1.08 grams / cm < ³ >. A method for separating a bodily fluid having many mixed fractions using a conventional centrifuge,
Centrifuging a device containing bodily fluids having a number of mixed fractions to form a first fraction, a second fraction and a third fraction of bodily fluids, and during the centrifuging step Using a valve means for controlling the ingress and egress of bodily fluid through the float inner isolation chamber of the device to isolate a third fraction within the float inner isolation chamber.   36. The method of claim 35, further comprising actuating a magnetic stir bar located within the float inner isolation chamber to agitate the third fraction after the centrifugation step.   36. The method of claim 35, further comprising collecting a third fraction of agitated body fluid from the device using an opening in open communication with the float inner isolation chamber. Prior to the centrifuging step, the method further comprises connecting at least one weight to the float to selectively adjust the float to a density of about 1.02 grams / cm ³ to about 1.08 grams / cm ^3. 36. The method of claim 35. A device for separating and isolating components of bodily fluids by centrifugation, said device comprising:
A storage container for storing body fluids having many components,
A float slidably disposed within the receiving container and having an inner isolation chamber, and the inner isolation chamber of the float for isolating at least one target component of bodily fluid within the inner isolation chamber of the float A device comprising valve means for controlling the entry and exit of body fluids through the body.   40. The method of claim 39, further comprising a magnetic stirring bar disposed inside the inner isolation chamber of the float for stirring at least one target component when the magnetic stirring bar is actuated by a magnetic field. apparatus.   40. The apparatus of claim 39, wherein the float includes opening means in open communication with the inner isolation chamber of the float for collecting at least one target component from the float. The float includes at least one weight to selectively adjust the density of the float prior to centrifugation to a density of about 1.02 grams / cm ³ to about 1.08 grams / cm ³ ; 40. The apparatus of claim 39.   The valve means includes first valve means for controlling the inflow of bodily fluid into the inner isolation chamber of the float and from the inner isolation chamber of the float. 40. The apparatus of claim 39, wherein the apparatus is for preventing fluid from flowing back through the valve means.   The valve means includes second valve means for controlling the outflow of body fluid to the outside of the inner isolation chamber of the float and for the outflow of body fluid to the second valve. 44. The apparatus of claim 43, wherein the apparatus is for preventing back flow through means into the inner isolation chamber of the float.

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