Systems, methods and components for isolating cells from a fluid sample

Abstract

A system for isolating preselected cell types from a fluid sample that includes a plurality of cell types includes a cell-capture fluidic chip, and a chip holder configured to receive the cell-capture fluidic chip and to maintain the cell-capture fluidic chip with a substantially fluid-tight seal while in operation. The chip holder is further configured to release the cell-capture fluidic chip to be removed from the chip holder for further processing. The cell-capture fluidic chip includes a substrate, a laser micro-dissection membrane disposed on the substrate, and a channel-defining layer disposed on the laser micro-dissection membrane. The laser micro-dissection membrane has a surface adapted to capture preselected cell types preferentially over other cell types of the plurality of cell types. The channel-defining layer is removable from the laser micro-dissection membrane for further processing of the cell-capture fluidic chip. 124

1 . A system for isolating preselected cell types from a fluid sample comprising a plurality of cell types, said system comprising:
 a cell-capture fluidic chip; and   chip holder configured to receive said cell-capture fluidic chip and to maintain said cell capture fluidic chip with a substantially fluid-tight seal while in operation,   wherein said chip holder is further configured to release said cell-capture fluidic chip to be removed from said chip holder for further processing,   wherein said cell-capture fluidic chip comprises:
 a substrate, 
 a laser micro-dissection membrane disposed on said substrate, and 
 a channel-defining layer disposed on said laser micro-dissection membrane, 
   wherein said laser micro-dissection membrane has a surface adapted to capture preselected cell types preferentially over other cell types of said plurality of cell types,   wherein said channel-defining layer is removable from said laser micro-dissection membrane for further processing of said cell-capture fluidic chip, and   wherein said cell-capture fluidic chip has an input port adapted to be connected to a fluid source and an output port to expel processed fluid such that said fluid sample flows from said fluid source through a fluid channel defined by said channel-defining layer over at least a portion of said surface of said laser micro-dissection membrane while in operation.   
     
     
         2 . A system for isolating preselected cell types according to  claim 1 , wherein said channel-defining layer defines said fluid channel such that at least a portion of said fluid channel has a chaotic mixing structure to cause at least partially turbulent flow. 
     
     
         3 . system for isolating preselected cell types according to  claim 1 , wherein said surface of said laser micro-dissection membrane has at least one of a structural or chemical modification to capture preselected cell types preferentially over other cell types of said plurality of cell types. 
     
     
         4 . A system for isolating preselected cell types according to  claim 3 , wherein said surface of said laser micro-dissection membrane comprises a chemical modification to capture circulating cancer cells (CTCs) preferentially over other cell types of said plurality of cell types. 
     
     
         5 . A system for isolating preselected cell types according to  claim 4 , wherein said surface of said laser micro-dissection membrane comprises a polymer. 
     
     
         6 . A system for isolating preselected cell types according to  claim 5 , wherein said polymer of said surface of said laser micro-dissection membrane comprises at least one of Poly(Lactic-co-Glycolic Acid) (PLGA), PolyCaproLactone (PCL), PolyLactide (PLA) or Chitosan (Poly-(D)glucosamine). 
     
     
         7 . A system for isolating preselected cell types according to  claim 6 , wherein said polymer further comprises polyethylene glycol (PEG). 
     
     
         8 . A system for isolating preselected cell types according to  claim 5 , wherein said polymer is at least partially formed into nano fibers. 
     
     
         9 . A system for isolating preselected cell types according to  claim 6 , wherein said polymer is at least partially formed into nano fibers. 
     
     
         10 . A system for isolating preselected cell types according to  claim 9 , wherein said polymer comprises a CTC capture agent attached thereto by at least one of biotin or streptavidin conjugation. 
     
     
         11 . A system for isolating preselected cell types according to  claim 10 , wherein said CTC capture agent comprises at least one of EpCAM, CA19-9, CD146, or CD147 antibodies. 
     
     
         12 . A system for isolating preselected cell types according to  claim 3 , wherein said surface of said laser micro-dissection membrane comprises a chemical modification to capture fetal nucleated red blood cells (fNRBCs) from maternal blood preferentially over other cell types of said plurality of cell types. 
     
     
         13 . A system for isolating preselected cell types according to  claim 12 , wherein said chemical modification comprises attaching at least CD71 and CD 147 antibodies to said surface of said laser micro-dissection membrane. 
     
     
         14 . A system for isolating preselected cell types according to  claim 1 , further comprising a fluid supply and control assembly adapted to be fluidly connected to said cell-capture fluidic chip while being held by said chip holder. 
     
     
         15 . A system for isolating preselected cell types according to  claim 14 , wherein said fluid supply and control assembly and said chip holder are mutually configured such that said chip holder is attachable to and removable from said fluid supply and control assembly while holding said cell-capture fluidic chip therein. 
     
     
         16 . A system for isolating preselected cell types according to  claim 1 , further comprising a laser that has a wavelength and power to cut said laser micro-dissection membrane at selected sections thereof to remove selected portions of said laser micro-dissection membrane for further processing. 
     
     
         17 . A system for isolating preselected cell types according to  claim 16 , wherein said laser is an ultraviolet laser. 
     
     
         18 . A system for isolating preselected cell types according to  claim 17 , further comprising an infrared laser and an infrared sensitive adhesion element,
 wherein said infrared sensitive adhesion element is adapted to be arranged proximate a laser dissected portion of said laser micro-dissection membrane such that said infrared sensitive adhesion element adheres thereto to facilitate removal of said laser dissected portion.   
     
     
         19 . A chip holder for maintaining a cell-capture fluidic chip with a substantially fluid-tight seal while in operation, comprising:
 a base component defining an indented region to accept said cell-capture fluidic chip therein;   a press component configured to be placed over said cell-capture fluidic chip; and a clamp assembly configured to clamp said base component and said press component together so as to maintain a substantially fluidly tight seal during operation,   wherein said clamp assembly is further configured to unclamp to release said cell-capture fluidic chip for removal from said chip holder.   
     
     
         20 . A chip holder according to  claim 19 , wherein said base component is adapted to be selectively attached to and removed from a fluid supply and control assembly. 
     
     
         21 . A chip holder according to  claim 20 , further comprising a turn-locking assembly configured to a user to increase pressure gradually. 
     
     
         22 . A chip holder according to  claim 21 , further comprising a plurality of springs arranged to equalize applied force across said cell-capture fluidic chip. 
     
     
         23 . A cell-capture fluidic chip for capturing preselected cell types from a fluid sample comprising a plurality of cell types, the cell-capture fluidic chip comprising:
 a substrate;   a laser micro-dissection membrane disposed on said substrate; and
 a channel-defining layer disposed on said laser micro-dissection membrane, 
   wherein said laser micro-dissection membrane has a surface adapted to capture preselected cell types preferentially over other cell types of said plurality of cell types,   wherein said channel-defining layer is removable from said laser micro-dissection membrane for further processing of said cell-capture fluidic chip, and   wherein said cell capture fluidic chip has an input port adapted to be connected to a fluid source and an output port to expel processed fluid such that said fluid sample flows from said fluid source through a fluid channel defined by said channel-defining layer over at least a portion of said surface of said laser micro-dissection membrane while in operation.   
     
     
         24 . A cell-capture fluidic chip according to  claim 23 , wherein said channel-defining layer defines said fluid channel such that at least a portion of said fluid channel has a chaotic mixing structure to cause at least partially turbulent flow. 
     
     
         25 . A cell-capture fluidic chip according to  claim 23 , wherein said surface of said laser micro-dissection membrane has at least one of a structural or chemical modification to capture preselected cell types preferentially over other cell types of said plurality of cell types. 
     
     
         26 . A cell-capture fluidic chip according to  claim 25 , wherein said surface of said laser micro-dissection membrane comprises a chemical modification to capture circulating cancer cells (CTCs) preferentially over other cell types of said plurality of cell types. 
     
     
         27 . A cell-capture fluidic chip according to  claim 26 , wherein said surface of said laser micro-dissection membrane comprises a polymer. 
     
     
         28 . A cell-capture fluidic chip according to  claim 27 , wherein said polymer of said surface of said laser micro-dissection membrane comprises at least one of Poly(Lactic-co-Glycolic Acid) (PLGA), PolyCaproLactone (PCL), PolyLactide (PLA) or Chitosan (Poly-(D)glucosamine). 
     
     
         29 . A cell-capture fluidic chip according to  claim 28 , wherein said polymer further comprises polyethylene glycol (PEG). 
     
     
         30 . A cell-capture fluidic chip according to  claim 27 , wherein said polymer is at least partially formed into nano fibers. 
     
     
         31 . A cell-capture fluidic chip according to  claim 28 , wherein said polymer is at least partially formed into nano fibers. 
     
     
         32 . A cell-capture fluidic chip according to  claim 31 , wherein said polymer comprises a CTC capture agent attached thereto by at least one of biotin or streptavidin conjugation. 
     
     
         33 . A cell-capture fluidic chip according to  claim 32 , wherein said CTC capture agent comprises at least one of EpCAM, CA19-9, CD146, or CD147 antibodies. 
     
     
         34 . A cell-capture fluidic chip according to  claim 25 , wherein said surface of said laser micro-dissection membrane comprises a chemical modification to capture fetal nucleated red blood cells (fNRBCs) from maternal blood preferentially over other cell types of said plurality of cell types. 
     
     
         35 . A cell-capture fluidic chip according to  claim 34 , wherein said chemical modification comprises attaching at least CD71 and CD 147 antibodies to said surface of said laser microdissection membrane. 
     
     
         36 . A method of isolating preselected cell types from a fluid sample comprising a plurality of cell types, comprising:
 providing a cell-capture fluidic chip, comprising:
 a substrate, 
 a laser micro-dissection membrane disposed on said substrate, and 
 a channel-defining layer disposed on said laser micro-dissection membrane, 
 wherein said laser micro-dissection membrane has a surface adapted to capture preselected cell types preferentially over other cell types of said plurality of cell types; 
   providing said fluid sample such that it flows through a fluid channel defined by said channel-defining layer over at least a portion of said surface of said laser micro-dissection membrane to capture preselected cell types from said fluid sample;   removing said channel-defining layer from said laser micro-dissection membrane and said substrate of said cell-capture fluidic chip after said providing said fluid sample; and   performing laser micro-dissection on said laser micro-dissection membrane and said substrate of said cell-capture fluidic chip after said removing said channel-defining layer to collect preselected cell types captured from said fluid sample.   
     
     
         37 . A method of isolating preselected cell types according to  claim 36 , wherein said performing laser micro-dissection comprises directing an ultraviolet laser onto said laser micro-dissection membrane to selectively cut off section of said laser micro-dissection membrane that have preselected cell types attached thereto. 
     
     
         38 . A method of isolating preselected cell types according to  claim 36 , wherein said removing said channel-defining layer from said laser micro-dissection membrane and said substrate comprises removing said cell-capture fluidic chip from a fluidly sealed chip holder.