Fluorescent Proteins: In Vivo Imaging
In vivo imaging is a powerful tool used to study individual plasmids or protein-protein interactions in deep-tissue organs and whole mammals. Genetically encoded fluorescent proteins most suitable for stable imaging of protein interactions in living cells and live mice are those with emission near or above 650 nm, as signals below 650 nm are highly absorbed by hemoglobin and cause undesirable background signals.
Many near-infrared fluorescent proteins for in vivo imaging are derived from bacterial phytochrome photoreceptors (BphPs). Spectrally distinct fluorescent iRFP variants have been shown to have high brightness and allow for single or multicolor imaging. Photo-activatable iRFPs can be turned on by non-phototoxic far-red light and can be used for spatially selective imaging of tissues in living animals. Further development of the original iRFP has resulted in a split fluorescence complementation probe used for detecting protein-protein interactions in vitro and in vivo. Learn more about in vivo imaging tools in this blog post.
Far Red
Protein | Excitation/Emission | Brightness | Plasmids |
---|---|---|---|
mCardinal | 604/659 | 18 | mCardinal plasmids from Chu et al. (2014) |
mNeptune | 600/650 | 13.4 | pcDNA3-mNeptune-N |
E2-Crimson | 611/646 | 28.98 | pEF.myc.ER-E2-Crimson |
Near-Infrared
Protein | Excitation/Emission | Brightness | Plasmids |
---|---|---|---|
iRFP670 | 643/670 | 12.7 | piRFP670-N1 |
miRFP670 | 642/670 | 12 | pmiRFP670-N1 |
iRFP682 | 663/682 | 10.2 | piRFP682-N1 |
iRFP702 | 673/702 | 7.6 | piRFP702-N1 |
miRFP703 | 673/703 | 8 | pmiRFP703-N1 |
miRFP709 | 683/709 | 4 | pmiRFP709-N1 |
iRFP (aka iRFP713) | 690/713 | 6.2 | piRFP |
iRFP720 | 702/720 | 5.8 | piRFP720-N1 |
iSplit | 690/713 | 5.3 | pPAS-E and pK-GAFm |
Photoactivatable
Protein | Excitation/Emission | Brightness | Plasmids |
---|---|---|---|
PAiRFP1 | 690/717 (after photoactivation) | 3.2 | pPAiRFP1-N1 |
PAiRFP2 | 692/719 (after photoactivation) | 3.0 | pPAiRFP2-N1 |
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