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Before In Vivo Imaging: Evaluation of Fluorescent Probes Using Fluorescence Microscopy, Multiplate Reader, and Cytotoxicity Assays

  • Shaojuan ZhangEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 1444)

Abstract

Fluorescent probes are widely utilized for noninvasive fluorescence imaging. Continuing efforts have been made in developing novel fluorescent probes with improved fluorescence quantum yield, enhanced target-specificity, and lower cytotoxicity. Before such probes are administrated into a living system, it is essential to evaluate the subcellular uptake, targeting specificity, and cytotoxicity in vitro. In this chapter, we briefly outline common methods used to evaluate fluorescent probes using fluorescence microscopy, multiplate reader, and cytotoxicity assay.

Key words

Fluorescence microscopy Multiplate reader Cytotoxicity Targeting specificity 

References

  1. 1.
    Han J, Burgess K (2010) Fluorescent indicators for intracellular pH. Chem Rev 110(5):2709–2728CrossRefPubMedGoogle Scholar
  2. 2.
    Quaranta M, Borisov SM, Klimant I (2012) Indicators for optical oxygen sensors. Bioanal Rev 4(2-4):115–157CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Wang X, Fang H, Huang Z, Shang W, Hou T, Cheng A, Cheng H (2013) Imaging ROS signaling in cells and animals. J Mol Med (Berl) 91(8):917–927CrossRefGoogle Scholar
  4. 4.
    Rao J, Dragulescu-Andrasi A, Yao H (2007) Fluorescence imaging in vivo: recent advances. Curr Opin Biotechnol 18(1):17–25CrossRefPubMedGoogle Scholar
  5. 5.
    Weissleder R, Tung CH, Mahmood U, Bogdanov A Jr (1999) In vivo imaging of tumors with protease-activated near-infrared fluorescent probes. Nat Biotechnol 17(4):375–378CrossRefPubMedGoogle Scholar
  6. 6.
    Sexton M, Woodruff G, Cudaback E, Kreitzer FR, Xu C, Lin YH, Möller T, Bai M, Manning HC, Bornhop D, Stella N (2009) Binding of NIR-conPK and NIR-6T to astrocytomas and microglial cells: evidence for a protein related to TSPO. PLoS One 4(12):e8271CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    van Meerloo J, Kaspers GL, Cloos J (2011) Cell sensitivity assays: the MTT assay. In: Cree IA (ed) Cancer cell culture, vol 731, Methods in molecular biology. Humana, New York, pp 237–245CrossRefGoogle Scholar
  8. 8.
    Zhang S, Shao P, Bai M (2013) In vivo type 2 cannabinoid receptor-targeted tumor optical imaging using a near infrared fluorescent probe. Bioconjug Chem 24(11):1907–1916CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Ling X, Zhang S, Shao P, Li W, Yang L, Ding Y, Xu C, Stella N, Bai M (2015) A novel near-infrared fluorescence imaging probe that preferentially binds to cannabinoid receptors CB2R over CB1R. Biomaterials 57:169–178CrossRefPubMedGoogle Scholar
  10. 10.
    Kosuge M, Takeuchi T, Nakase I, Jones AT, Futaki S (2008) Cellular internalization and distribution of arginine-rich peptides as a function of extracellular peptide concentration, serum, and plasma membrane associated proteoglycans. Bioconjug Chem 19(3):656–664CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Molecular Imaging Lab, Department of RadiologyUniversity of PittsburghPittsburghUSA

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