High-density microcavity array for cell detection: single-cell analysis of hematopoietic stem cells in peripheral blood mononuclear cells.
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 single-cell 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.
Hosokawa
Masahito
M
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Arakaki
Atsushi
A
Takahashi
Masayuki
M
Mori
Tetsushi
T
Takeyama
Haruko
H
Matsunaga
Tadashi
T
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism*; D002469:Cell Separation; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D046888:Tissue Array Analysis / Q000379:methods*
D000906:Antibodies; D018952:Antigens, CD34