QDot nanocrystals in intracellular flow: ZAP-70, Ki-67 and five-colors fluorophores panel

Ilya Sotnikov, Shulamit Jaron, Marina Konopleva, Susan O'Brien, Michael Andreeff, Julia Hillabrant, John Manis, Jennifer Brown, Ivan Vorobjev, Natasha Barteneva

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

Abstract

We present for the first time a methodological approach for the measurement of cytoplasmic Zap-70 and nuclear Ki-67 proteins with QDot nanocrystal-labeled antibodies. Mononuclear cells from chronic lymphoid leukemia patients were used to detect Zap-70, and Jurkat T-cells for Ki-67. Intracellular staining kits based on saponin and paraformaldehyde were chosen as optimal for the delivery of QDot nanocrystals inside cells. Single staining with QDot 565-, 605-, or 655-labeled antibodies is operative in intracellular flow cytometry. In multicolor panels, the most robust results are obtained with the QDot 655 detected in the APC channel (633 nm excitation, 660/20 nm band pass or customized 655/20 filter) to simplify the spectral overlap compensation. In 3-, 4-, and 5-color combinations signal brightness for intracellular QDot 655 does not decrease, whereas positive-to-negative signal ratio is higher than with organic dyes. The optimized combination of fluorophores for 5-color panel includes APC-Cy7, PE, PE-Cy7, FITC, and QDot 655. We used CD3, CD19, CD38, and CD5 surface markers. Intracellular QDots outperformed organic fluorophores probed in the same samples. QDot nanocrystals are a robust tool in intracellular flow cytometry due to their narrow emission spectrum, brightness, and stability of this fluorophore.

Original languageEnglish
Title of host publicationNanotechnology 2010
Subtitle of host publicationBio Sensors, Instruments, Medical, Environment and Energy - Technical Proceedings of the 2010 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2010
Pages424-427
Number of pages4
Publication statusPublished - Nov 9 2010
Externally publishedYes
EventNanotechnology 2010: Bio Sensors, Instruments, Medical, Environment and Energy - 2010 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2010 - Anaheim, CA, United States
Duration: Jun 21 2010Jun 24 2010

Publication series

NameNanotechnology 2010: Bio Sensors, Instruments, Medical, Environment and Energy - Technical Proceedings of the 2010 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2010
Volume3

Other

OtherNanotechnology 2010: Bio Sensors, Instruments, Medical, Environment and Energy - 2010 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2010
CountryUnited States
CityAnaheim, CA
Period6/21/106/24/10

Keywords

  • Cytometry
  • Ki-67
  • Quantum dots
  • ZAP-70

ASJC Scopus subject areas

  • Bioengineering
  • Biomedical Engineering
  • Electrical and Electronic Engineering

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