Multiplexed living cells stained with quantum dot bioprobes for multiplexed detection of single-cell array

Abstract. The results of quantum dot (QD) probe preparation for multiplexed single-cell array staining and analysis are reported. By controlling the reaction temperature, time, and ratio of Cd to Se, multicolor CdSe QDs emitting fluorescence ranging from purple to red in a safer, simpler, and more convenient way than traditional methods is obtained. To detect cells using these oil-soluble QDs, they are first coated with water-soluble thioglycolic aid (TGA) so that biocompatible multiwavelength bioprobes can be obtained. QDs’ surface is somewhat damaged when binding TGA to QDs is found, which results in a reduction of QDs’ emission wavelength and a slight blue shift of QDs’ emission wavelength after water-soluble modification with TGA. Comparison of the emission spectrum showed that it is negligible, and the fluorescent properties of QDs capped by TGA are still satisfactory. Living cells are then stained with multiplexed probes by conjugating TGA-QDs with antibodies specific to these cell antigens. Changes in fluorescence intensity can indicate change in the relative quantity of antigens expressed in the same cell caused by external stimulus, offering effective methods to multiplexed optical analysis of single cells.

[1]  P. Mulvaney,et al.  The effects of chemisorption on the luminescence of CdSe quantum dots. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[2]  C. Hsu,et al.  Over‐expression of nucleophosmin/B23 decreases the susceptibility of human leukemia HL‐60 cells to retinoic acid‐induced differentiation and apoptosis , 2000, International journal of cancer.

[3]  Vicki L. Colvin,et al.  X-ray Photoelectron Spectroscopy of CdSe Nanocrystals with Applications to Studies of the Nanocrystal Surface , 1994 .

[4]  F. Lund-Johansen,et al.  Expression of leukocyte differentiation antigens during the differentiation of HL‐60 cells induced by 1,25‐dihydroxyvitamin D3: comparison with the maturation of normal monocytic and granulocytic bone marrow cells , 1995, Journal of leukocyte biology.

[5]  Li Cao,et al.  A new route to zinc-blende CdSe nanocrystals: mechanism and synthesis. , 2005, The journal of physical chemistry. B.

[6]  T. Thepen,et al.  Cytotoxicity of anti-CD64-ricin a chain immunotoxin against human acute myeloid leukemia cells in vitro and in SCID mice. , 2001, Journal of hematotherapy & stem cell research.

[7]  L. Balan,et al.  Folic acid-conjugated core/shell ZnS:Mn/ZnS quantum dots as targeted probes for two photon fluorescence imaging of cancer cells. , 2011, Acta biomaterialia.

[8]  Qiang Wang,et al.  Luminescent properties of water-soluble denatured bovine serum albumin-coated CdTe quantum dots. , 2006, The journal of physical chemistry. B.

[9]  Igor Nabiev,et al.  Energy Transfer in Aqueous Solutions of Oppositely Charged CdSe/ZnS Core/Shell Quantum Dots and in Quantum Dot−Nanogold Assemblies , 2004 .

[10]  Z. You,et al.  A Single Cells Patterning Approach for Human Promyelocytic Leukemia Cells , 2013 .

[11]  R. Misra.  Quantum dots for tumor-targeted drug delivery and cell imaging. , 2008, Nanomedicine.

[12]  Chih-Ming Ho,et al.  Photolithographic patterning of organosilane monolayer for generating large area two-dimensional B lymphocyte arrays. , 2008, Lab on a chip.

[13]  K. Schwarz,et al.  Breaking chemoresistance and radioresistance with [213Bi]anti-CD45 antibodies in leukemia cells. , 2007, Cancer research.

[14]  M. Bawendi,et al.  Synthesis and characterization of nearly monodisperse CdE (E = sulfur, selenium, tellurium) semiconductor nanocrystallites , 1993 .

[15]  Z. Jia,et al.  Tunable ZnO quantum dots for bioimaging: synthesis and photoluminescence , 2013 .

[16]  Weidong Yang,et al.  Shape control of CdSe nanocrystals , 2000, Nature.

[17]  S. Nie,et al.  Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules , 2001, Nature Biotechnology.

[18]  Daniele Gerion,et al.  Fluorescent CdSe/ZnS nanocrystal-peptide conjugates for long-term, nontoxic imaging and , 2004 .

[19]  Lehui Lu,et al.  Fluorescence-enhanced gadolinium-doped zinc oxide quantum dots for magnetic resonance and fluorescence imaging. , 2011, Biomaterials.

[20]  Xiaogang Peng,et al.  Kinetics of II-VI and III-V Colloidal Semiconductor Nanocrystal Growth: “Focusing” of Size Distributions , 1998 .

[21]  S. Pathak,et al.  Hydroxylated quantum dots as luminescent probes for in situ hybridization. , 2001, Journal of the American Chemical Society.

[22]  Tianhai Tian,et al.  The origins of cancer robustness and evolvability. , 2011, Integrative biology : quantitative biosciences from nano to macro.

[23]  Franziska Michor,et al.  Evolution of Resistance to Targeted Anti-Cancer Therapies during Continuous and Pulsed Administration Strategies , 2009, PLoS Comput. Biol..

[24]  S. Nie,et al.  In vivo cancer targeting and imaging with semiconductor quantum dots , 2004, Nature Biotechnology.

[25]  Jackie Y Ying,et al.  Synthesis of carbohydrate-conjugated nanoparticles and quantum dots. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[26]  Hao Zhang,et al.  Studies on quantum dots synthesized in aqueous solution for biological labeling via electrostatic interaction. , 2003, Analytical biochemistry.

[27]  M. Steigerwald,et al.  Formation of metal-chalcogen bonds by the reaction of metal-alkyls with silyl chalcogenides , 1989 .

[28]  Xiaogang Peng,et al.  Formation of high-quality CdTe, CdSe, and CdS nanocrystals using CdO as precursor. , 2001, Journal of the American Chemical Society.

[29]  S. Nie,et al.  Quantum dot bioconjugates for ultrasensitive nonisotopic detection. , 1998, Science.

[30]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[31]  Peng Zhang,et al.  ZnO QD@PMAA-co-PDMAEMA nonviral vector for plasmid DNA delivery and bioimaging. , 2010, Biomaterials.

[32]  Vincent Noireaux,et al.  In Vivo Imaging of Quantum Dots Encapsulated in Phospholipid Micelles , 2002, Science.

[33]  R. Westervelt,et al.  Incorporation of iron oxide nanoparticles and quantum dots into silica microspheres. , 2008, ACS nano.

[34]  Liwei Lin,et al.  Quantum dot nano thermometers reveal heterogeneous local thermogenesis in living cells. , 2011, ACS nano.

[35]  Xiaohu Gao,et al.  Quantum dot-amphipol nanocomplex for intracellular delivery and real-time imaging of siRNA. , 2008, ACS nano.

[36]  R. Misra,et al.  New generation of chitosan-encapsulated ZnO quantum dots loaded with drug: synthesis, characterization and in vitro drug delivery response. , 2010, Acta biomaterialia.

[37]  Z. Rosenzweig,et al.  Luminescent CdSe Quantum Dot Doped Stabilized Micelles , 2002 .