The Advantages of Using Fluorescent Proteins for In Vivo Imaging

Robert M. Hoffman1

1 AntiCancer Inc., San Diego
Publication Name:  Current Protocols Essential Laboratory Techniques
Unit Number:  Unit 9.6
DOI:  10.1002/cpet.12
Online Posting Date:  November, 2017
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Abstract

Fluorescent proteins have revolutionized biology by enabling what was formerly invisible to be seen clearly, and potentially enable imaging of all cells in living animals. There are numerous applications for in vivo imaging with fluorescent proteins, including use in cancer, immunology, infection, and neurobiology studies. This unit describes how introducing the GFP and RFP gene into malignant tissue has enabled imaging of primary cancer and the metastatic process with subcellular resolution in vivo. © 2017 by John Wiley & Sons, Inc.

Keywords: tumor; orthotopic; cancer cells; GFP; RFP; CFP; imaging; metastasis; CTC; bacterial tumor‐targeting; fluorescence‐guided surgery; stem cells; nude mice

     
 
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Table of Contents

  • Overview and Principles
  • The In Vivo GFP Revolution
  • Techniques
  • Basic Protocol 1: Whole‐Body Imaging with a Flashlight
  • Why Fluorescent Proteins are the Method of Choice for Whole‐Body Imaging
  • Definitions
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Whole‐Body Imaging with a Flashlight

  Materials
  • Plant, animal, or bacterial sample expressing fluorescent protein (GFP, YFP, or RFP)
  • Anesthesia as needed (animals)
  • A blue LED flashlight (variety of sources: Amazon Crime Scene Blue LED Flashlight 455 460 nm; Nightsea flashlight (http://www.nightsea.com); or Mecan flashlight (http://www.mecanusa.com/led/led‐flashlight‐RH3w.htm) with an excitation filter (midpoint wavelength peak of 460 to 480 nm) attached to front of flashlight and a emission filter 510 to 530 (Omega Filters; Chroma Technology) mounted in front of the lens of the digital camera (on tripod), cell phone, or CCD imaging system used for gels and blots—for a good description of a homemade blue LED GFP/RFP tool, see http://www.wormbook.org/wbg/articles/volume‐18‐number‐1/using‐leds‐as‐a‐low‐cost‐source‐to‐detect‐gfp‐and‐dsred‐2/
  • Digital SLR or CCD/CMOS‐based in vivo imaging system (unit 7.5; Moomaw, Medberry, & Gallagher, )
  • Windowless dark room, benchtop darkroom, or imaging cabinet
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Figures

Videos

Literature Cited

Literature Cited
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