Autor: |
Jie Liu, Radoslaw Bombera, Loïc Leroy, Yoann Roupioz, Dieudonné R Baganizi, Patrice N Marche, Vincent Haguet, Pascal Mailley, Thierry Livache |
Jazyk: |
angličtina |
Rok vydání: |
2013 |
Předmět: |
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Zdroj: |
PLoS ONE, Vol 8, Iss 3, p e57717 (2013) |
Druh dokumentu: |
article |
ISSN: |
1932-6203 |
DOI: |
10.1371/journal.pone.0057717 |
Popis: |
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. METHODOLOGY/PRINCIPAL FINDINGS: 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/SIGNIFICANCE: 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. |
Databáze: |
Directory of Open Access Journals |
Externí odkaz: |
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