pp 1-35 | Cite as

Applications of Fluorescent Protein-Based Sensors in Bioimaging

  • Uday Kumar Sukumar
  • Arutselvan Natarajan
  • Tarik F. Massoud
  • Ramasamy PaulmuruganEmail author
Part of the Topics in Medicinal Chemistry book series


In the last two decades, there have been enormous developments in the area of reporter gene imaging for various bioimaging applications, especially to track cellular events that are occurring in intact cells and cells within living animals. As part of this process, there has been a significant interest in identifying new reporters or developing new substrates that can allow us to image multiple cellular events simultaneously without any signal overlap between the targets. Even though chemical dyes are useful for some of these applications, reporter proteins which mimic biological properties of proteins when tagged directly with the target proteins are very useful. Although molecular imaging has significantly advanced through use of different imaging probes (radiolabeled ligands, MR contrast agents, CT contrast agents, fluorescent dyes, fluorescent and bioluminescent proteins) and techniques (PET, SPECT, MRI, CT, optical, ultrasound, and photoacoustic imaging), optical imaging, such as fluorescence and bioluminescence imaging, has shown promising applications in various preclinical settings, especially in imaging cellular pathways and studies involving drug development. This is mainly owing to its simple and easy nature in performing the assay and also its high-throughput and cost-effective applications. In this chapter, we review the evolution of optical imaging with specific emphasis on fluorescent proteins, as well as an introduction regarding the general approach of optical imaging in in vitro and in vivo applications. We explain this by briefly introducing different optical imaging methods and fluorescent assays developed based on fluorescent dyes and fluorescent proteins followed by a detailed review of different fluorescent proteins currently used for various assay developments and applications.


BRET Fluorescence dyes Fluorescent proteins FRET In vivo imaging 



We would like to thank the Canary Center at Stanford, Department of Radiology, for providing facility and resources. We also thank SCi3 Small Animal Imaging Service Center, Stanford University School of Medicine, for providing imaging facilities and data analysis support. We acknowledge Dr. Sanjiv Sam Gambhir, Chair of the Department of Radiology, Stanford University, for his constant support and help.

Compliance with Ethical Standards

Conflicts of Interest

There are no actual or potential conflicts of interest in regard to this chapter.


This research was supported by NIH R01CA209888 and NIH R21EB022298. This work was also in part supported by the Center for Cancer Nanotechnology Excellence for Translational Diagnostics (CCNE-TD) at Stanford University through an award (grant no. U54 CA199075) from the National Cancer Institute (NCI) of the National Institutes of Health (NIH).

Ethical Approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.


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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Uday Kumar Sukumar
    • 1
  • Arutselvan Natarajan
    • 1
  • Tarik F. Massoud
    • 1
  • Ramasamy Paulmurugan
    • 1
    Email author
  1. 1.Molecular Imaging Program at Stanford, Department of RadiologyStanford University School of MedicineStanfordUSA

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