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Support >  Applications >  Using_Fluorescent_Proteins >  In_Vivo_Imaging_with_RFP

In Vivo Imaging with Red Fluorescent Proteins

In Vivo Imaging with Red Fluorescent Proteins

Application: Noninvasive Optical Tracking of Breast Cancer Metastasis with tdTomato Red Fluorescent Protein

tdTomato’s emission wavelength (581 nm) and brightness make it ideal for live animal imaging studies. In one xenograft mouse model of metastatic breast cancer, tdTomato was easily detected as deep as 1 cm below the surface, and extremely small lesions could be detected and tracked over time (Figure 1; 1).

Read more about in vivo imaging with tdTomato

Cells labeled with tdTomato are detected 1 cm deep in a SCID mouse model.

tdTomato, but not GFP, can be detected in the SCID mouse cadaver phantom model. False-color overlay images (regions of interest encircled) indicate that the imaging system could detect tdTomato fluorescence in the cadaver model, but not GFP fluorescence. Panel A. Implanted tube with 100 x 106 MDA-MB-231-tdTomato-expressing cells, imaged with the DsRed filter set. Exposure time: 1 sec. Panel B. Implanted tube with 100 x 106 MDA-MB-231-GFP-expressing cells, imaged with the GFP filter set. Exposure time: 1 sec.

Red and far red fluorescent proteins are ideal for in vivo imaging due to their reduced autofluorescence. We recommend the following fluorescent proteins for plant and animal imaging studies:

Far Red Fluorescent Proteins

  • mPlum was developed in Dr. Roger Tsien’s lab by directed mutagenesis of mRFP1, a monomeric mutant of DsRed (2–5).
  • E2-Crimson matures quickly, and has high photostability, high solubility, and low cytotoxicity. It was designed was designed specifically for in vivo and stem cell applications (6).
  • HcRed is a far-red fluorescent protein derived from a nonfluorescent chromoprotein found in the Anthozoa-class sea anemone Heteractis crispa. HcRed1 has a low tendency to form aggregates in living cells.

Red Fluorescent Proteins

  • mCherry has been successfully fused to many proteins and widely used for quantitative imaging techniques including fluorescence resonance energy transfer (FRET). mCherry was developed in Dr. Roger Tsien’s lab by directed mutagenesis of mRFP1, a monomeric mutant of DsRed (2–5).
  • tdTomato (also developed in Dr. Roger Tsien’s lab) is a genetic fusion of two copies of the dTomato gene (5) which was specifically designed for low aggregation. Its tandem dimer structure plays an important role in the exceptional brightness of tdTomato. tdTomato can be seen in SCID mice, as deep as 1 cm below the surface (7). tdTomato has also been used to quantify breast cancer tumor growth in response to target gene activation (1). Transgenic mouse models have also been developed, including one where tdTomato was used as a reporter for Cre recombination. This model was also useful as a tool for cell lineage tracing, transplantation studies, and analysis of cell morphology in vivo (8).
  • DsRed-Express2 has accelerated maturation and high solubility, and was designed specifically for in vivo and stem cell applications (9).
Fluorescent Protein Excitation Maxima (nm) Emission Maxima (nm)
Far Red
    mPlum 590 649
    E2-Crimson 611 646
    HcRed1 588 681
Red
    mCherry 587 610
    tdTomato 554 581
    DsRed-Express2 554 591

References

  1. Winnard Jr., P. T. et al. (2006) Neoplasia 8(10):796–806.
  2. Shaner, N. C. et al. (2004) Nature Biotechnol. 22(12):1567–1572.
  3. Wang, L. et al. (2004) Proc. Nat. Acad. Sci. USA 101(48):16745–16749.
  4. Shu, X. et al. (2006) Biochemistry 45(32):9639–9647.
  5. Campbell, R. E. et al. (2002) Proc. Nat. Acad. Sci. USA 99(12):7877–7882.
  6. Strack, R. L. et al. (2009) Biochemistry 48(35):8279–8281.
  7. Johnstone, C. N. et al. (2008) Mol. Cell Biol. 28(2):687–704.
  8. Muzumdar, M. D. et al. (2007) Genesis 45(9):593–605.
  9. Strack, R. L. et al. (2008) Nat Methods. 5(11):955–957.
 
Products
Cat. # Product Package Size Price License # of Units Select
632410 pHcRed1 Vector 20 ug $574.00 License Statements *
632411 pHcRed1-1 Vector 20 ug $574.00 License Statements *
632415 pHcRed1-C1 Vector 20 ug $574.00 License Statements *
632422 pHcRed1-DR Vector 20 ug $574.00 License Statements *
632522 pmCherry Vector 20 ug $516.00 License Statements *
632523 pmCherry-N1 Vector 20 ug $516.00 License Statements *
632524 pmCherry-C1 Vector 20 ug $516.00 License Statements *
632525 pmCherry-1 Vector 20 ug $516.00 License Statements *
632527 pmPlum Vector 20 ug $516.00 License Statements *
632531 ptdTomato Vector 20 ug $516.00 License Statements *
632532 ptdTomato-N1 Vector 20 ug $516.00 License Statements *
632533 ptdTomato-C1 Vector 20 ug $516.00 License Statements *
632534 pCMV-tdTomato Vector 20 ug $516.00 License Statements *
632190 DD-tdTomato Reporter System Each $569.00 License Statements *
632535 pDsRed-Express2 Vector 20 ug $557.00 License Statements *
632536 pDsRed-Express2-1 Vector 20 ug $557.00 License Statements *
632537 pDsRed-Express2-N1 Vector 20 ug $557.00 License Statements *
632538 pDsRed-Express2-C1 Vector 20 ug $557.00 License Statements *
632539 pCMV DsRed-Express2 Vector 20 ug $557.00 License Statements *
632540 pIRES2 DsRed-Express2 Vector 20 ug $557.00 License Statements *
631237 pLVX-IRES-mCherry Vector 20 ug $676.00 License Statements *
631238 pLVX-IRES-tdTomato Vector 20 ug $676.00 License Statements *
632553 pE2-Crimson Vector 20 ug $574.00 License Statements *
632554 pE2-Crimson-N1 Vector 20 ug $574.00 License Statements *
632556 pCMV-E2-Crimson Vector 20 ug $574.00 License Statements *
632555 pE2-Crimson-C1 Vector 20 ug $574.00 License Statements *
632559 pLVX-DsRed-Express2-C1 Vector 10 ug $676.00 License Statements *
632560 pLVX-DsRed-Express2-N1 Vector 10 ug $676.00 License Statements *
632561 pLVX-mCherry-C1 Vector 10 ug $676.00 License Statements *
632562 pLVX-mCherry-N1 Vector 10 ug $676.00 License Statements *
632563 pLVX-tdTomato-N1 Vector 10 ug $676.00 License Statements *
632564 pLVX-tdTomato-C1 Vector 10 ug $676.00 License Statements *
632567 pRetroQ-mCherry-C1 Vector 10 ug $676.00 License Statements *
632568 pRetroQ-mCherry-N1 Vector 10 ug $676.00 License Statements *
631078 pLVX-mCherry-Actin Vector 10 ug $676.00 License Statements *
 
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