title: Selective individual primary cell capture using locally bio-functionalized micropores.
abstract: BACKGROUND
  Solid-state micropores have been widely employed for 6 decades to recognize and size flowing unlabeled cells. However, the resistive-pulse technique presents limitations when the cells to be differentiated have overlapping dimension ranges such as B and T lymphocytes. An alternative approach would be to specifically capture cells by solid-state micropores. Here, the inner wall of 15-µm pores made in 10 µm-thick silicon membranes was covered with antibodies specific to cell surface proteins of B or T lymphocytes. The selective trapping of individual unlabeled cells in a bio-functionalized micropore makes them recognizable just using optical microscopy.
  
  
  RESULTS
  We locally deposited oligodeoxynucleotide (ODN) and ODN-conjugated antibody probes on the inner wall of the micropores by forming thin films of polypyrrole-ODN copolymers using contactless electro-functionalization. The trapping capabilities of the bio-functionalized micropores were validated using optical microscopy and the resistive-pulse technique by selectively capturing polystyrene microbeads coated with complementary ODN. B or T lymphocytes from a mouse splenocyte suspension were specifically immobilized on micropore walls functionalized with complementary ODN-conjugated antibodies targeting cell surface proteins.
  
  
  CONCLUSIONS
  The results showed that locally bio-functionalized micropores can isolate target cells from a suspension during their translocation throughout the pore, including among cells of similar dimensions in complex mixtures.
authors.affiliation: Institut Nanosciences et Cryogénie, UMR5819 CEA/CNRS/UJF, Grenoble, France.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.forename: Radoslaw
authors.initials: R
authors.lastname: Bombera
authors.forename: Loïc
authors.initials: L
authors.lastname: Leroy
authors.forename: Yoann
authors.initials: Y
authors.lastname: Roupioz
authors.forename: Dieudonné R
authors.initials: DR
authors.lastname: Baganizi
authors.forename: Patrice N
authors.initials: PN
authors.lastname: Marche
authors.forename: Vincent
authors.initials: V
authors.lastname: Haguet
authors.forename: Pascal
authors.initials: P
authors.lastname: Mailley
authors.forename: Thierry
authors.initials: T
authors.lastname: Livache
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology*; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D005374:Filtration / Q000295:instrumentation / Q000379:methods; D018796:Immunoconjugates / Q000737:chemistry; D051379:Mice; D008863:Microspheres; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D011758:Pyrroles / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry; D013154:Spleen / Q000166:cytology*; D013601:T-Lymphocytes / Q000166:cytology*
chemical_list: D000906:Antibodies; D018796:Immunoconjugates; D009838:Oligodeoxyribonucleotides; D011108:Polymers; D011758:Pyrroles; C067635:polypyrrole; D012822:Silicon Dioxide