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Qdot ® nanocrystal conjugates to expand the fluor palette for polychromatic flow cytometry
Author(s) -
Godfrey William L.,
Zhang YuZhong,
Jaron Shulamit,
Buller Gayle M.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.567
Subject(s) - nanocrystal , materials science , quantum dot , flow cytometry , wavelength , conjugate , optoelectronics , nanotechnology , microbiology and biotechnology , biology , mathematical analysis , mathematics
Qdot nanocrystal conjugates against leukocyte surface markers provide powerful tools to multiply fluorophore selection in flow cytometry. These semiconductor nanocrystals are best excited by UV or violet light, and provide symmetrical emission peaks 150 to 400 nm above their excitation wavelengths. Care is required with filter selection and attention to cross‐laser compensation for spectral overlap. In this study, we demonstrate Qdot conjugate compatibility with standard sample preparation reagents for fixation, permeabilization, and erythrocyte lysis. We optimize filter combinations for Qdot conjugates and conventional fluors. Studies were performed on human peripheral blood leukocytes using a BD LSR II flow cytometer. Filters were optimized for nanocrystal / fluor combinations, with attention to signal intensity and compensation. For example, compensation of Qdot 605 nanocrystal emission out of the RPE‐Texas Red ® dye channel was reduced from ~100% to ~45% by changing from a 610 to a 620 band pass filter. As a result of individual optimizations, 8 to 10 color experiments have been run using 2 to 4 Qdot nanocrystal conjugates with good resolution of cell populations. Due to the unique spectral characteristics of nanocrystals, Qdot nanocrystal conjugates can dramatically facilitate multicolor experimental design when performing flow cytometry experiments. Supported by Invitrogen Corporation.