TY - JOUR
T1 - High-density microcavity array for cell detection
T2 - Single-cell analysis of hematopoietic stem cells in peripheral blood mononuclear cells
AU - Hosokawa, Masahito
AU - Arakaki, Atsushi
AU - Takahashi, Masayuki
AU - Mori, Tetsushi
AU - Takeyama, Haruko
AU - Matsunaga, Tadashi
PY - 2009/7/1
Y1 - 2009/7/1
N2 - Detection and isolation of specific cell types from limited biological samples have become a major challenge in clinical diagnosis and cell biology research. Here, we report a high-density microcavity array for target cell detection in which thousands of single cells were neatly arrayed onto 10 000 microcavities with high efficiency at approximately 90% of the loaded cells. Cell-specific immunophenotypes were exclusively identified at the singlecell level by measuring fluorescence intensities of cells labeled with antibodies targeting cell surface markers, and the purity of hematopoietic stem cells (HSCs) within human peripheral blood analyzed by this system was correlated with those obtained by conventional flow cytometry. Furthermore, gene expression of the stem cell marker, CD34, was determined from HSCs by isolating single cells using a micromanipulator. This technology has proven to be an effective tool for target cell detection and subsequent cellular analytical research at the single-cell level.
AB - Detection and isolation of specific cell types from limited biological samples have become a major challenge in clinical diagnosis and cell biology research. Here, we report a high-density microcavity array for target cell detection in which thousands of single cells were neatly arrayed onto 10 000 microcavities with high efficiency at approximately 90% of the loaded cells. Cell-specific immunophenotypes were exclusively identified at the singlecell level by measuring fluorescence intensities of cells labeled with antibodies targeting cell surface markers, and the purity of hematopoietic stem cells (HSCs) within human peripheral blood analyzed by this system was correlated with those obtained by conventional flow cytometry. Furthermore, gene expression of the stem cell marker, CD34, was determined from HSCs by isolating single cells using a micromanipulator. This technology has proven to be an effective tool for target cell detection and subsequent cellular analytical research at the single-cell level.
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U2 - 10.1021/ac900535h
DO - 10.1021/ac900535h
M3 - Article
C2 - 19485404
AN - SCOPUS:67649933548
SN - 0003-2700
VL - 81
SP - 5308
EP - 5313
JO - Analytical chemistry
JF - Analytical chemistry
IS - 13
ER -