Development of a microfabricated cytometry platform for characterization and sorting of individual leukocytes

Alexander Revzin, Kazuhiko Sekine, Aaron Sin, Ronald G. Tompkins, Mehmet Toner

Research output: Contribution to journalArticle

139 Citations (Scopus)

Abstract

Organizing leukocytes into high-density arrays makes these cells amenable to rapid optical characterization and subsequent sorting, pointing to clinical and basic science applications. The present paper describes development of a cytometry platform for creating high-density leukocyte arrays and demonstrates retrieval of single cells from the array. Poly(ethylene glycol) (PEG) photolithography was employed to fabricate arrays of microwells composed of PEG hydrogel walls and glass attachment pads 20 μ × 20 μm and 15 μm × 15 μ in size. PEG micropatterned glass surfaces were further modified with cell-adhesive ligands, poly-L-lysine, anti-CD5 and anti-CD 19 antibodies, in order to engineer specific cell-surface interactions within the individual wells. Localization of the fluorescently-labeled proteins in the glass attachment pads of PEG microwells was visualized by fluorescence microscopy. Glass slides micropatterned with PEG and cell-adhesive ligands were exposed to T-lymphocytes for 30 min. These anchorage-independent cells became selectively captured in the ligand-modified microwells forming high-density cell arrays. Cell occupancy in the microwells was found to be antibody-dependent, reaching 94.6 ± 2.3% for microwells decorated with T-cell specific anti-CD5 antibodies. Laser capture microdissection (LCM) was investigated as a method for sorting cells from the array and retrieval of single selected cells was demonstrated.

Original languageEnglish (US)
Pages (from-to)30-37
Number of pages8
JournalLab on a Chip
Volume5
Issue number1
DOIs
StatePublished - Jan 1 2005
Externally publishedYes

Fingerprint

Sorting
Polyethylene glycols
Leukocytes
Antibodies
Glass
T-cells
Ligands
Adhesives
Microdissection
Hydrogel
Fluorescence microscopy
Photolithography
Laser Capture Microdissection
T-Lymphocytes
Hydrogels
Lysine
Ethylene Glycol
Fluorescence Microscopy
Cell Communication
Proteins

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Chemistry(all)
  • Biomedical Engineering

Cite this

Development of a microfabricated cytometry platform for characterization and sorting of individual leukocytes. / Revzin, Alexander; Sekine, Kazuhiko; Sin, Aaron; Tompkins, Ronald G.; Toner, Mehmet.

In: Lab on a Chip, Vol. 5, No. 1, 01.01.2005, p. 30-37.

Research output: Contribution to journalArticle

Revzin, Alexander ; Sekine, Kazuhiko ; Sin, Aaron ; Tompkins, Ronald G. ; Toner, Mehmet. / Development of a microfabricated cytometry platform for characterization and sorting of individual leukocytes. In: Lab on a Chip. 2005 ; Vol. 5, No. 1. pp. 30-37.
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