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Support >  Applications >  Using_Fluorescent_Proteins >  In_Vivo_Imaging_with_GFP

Green Fluorescent Proteins for In Vivo Imaging

Green Fluorescent Proteins for In Vivo Imaging

We recommend ZsGreen1 and AcGFP1 for in vivo imaging studies.

Fluorescent Protein Excitation Maxima (nm) Emission Maxima (nm)
ZsGreen1 493 505
AcGFP1 475 505

Application: Green Fluorescent Proteins in Plant Research

Plants and plant tissues can be successfully imaged with green fluorescent proteins. Although many plants contain endogenous compounds such as phenolics that fluoresce in the green to yellow range with low excitation wavelengths (1–2), the use of proper controls eliminates the problems associated with this autofluorescence.

Read more about Fluorescent Proteins in Plant Biotechnology

Soybean expressing ZsGreen green fluorescent protein.

Figure 1. Expression of ZsGreen in soybean under white light (Panel A) and fluorescent light (Panel B). Bar=1 cm.

Maize expressing AcGFP green fluorescent protein.

Figure 2. AcGFP expression in maize tissues and plants. Panels A and D. Maize callus under white light and fluorescent light. Panels B and E. Transformed maize leaf and root under white light and fluorescent light. Panels C and F. Kernels from a segregating transformed line under white light and fluorescent light. Imaging used a Zeiss Sv-11 dissecting scope, HB100 light source, and a GFP (plant) cube; exciter HQ470/40, dichroic 495LP, emission D525/50 BP.

Application: ZsGreen Labeling of Breast Cancer Cells to Visualize Metastasis

The brightness and enhanced ability of ZsGreen to withstand fixation makes it an ideal fluorescent protein for in vivo metastasis studies. ZsGreen was used to label human estrogen receptor-positive breast cancer cells, and fluorescent whole body imaging on living, non-anesthetized, immunocompromised nude mice identified lymph node metastases that arose from estrogen-dependent tumors grown orthotopically in mammary glands. Metastatic cancer cells injected directly into the circulation were detected in bone, brain, adrenal gland, and lung by fluorescence imaging (Figures 3–4; 3).

Read more about in vivo imaging with ZsGreen1

Breast cancer cells expressing ZsGreen1 were visualized with fluorescent whole body imaging.

Figure 3. Fluorescent whole body image of a mouse with bilateral solid tumors derived from estrogen receptor-positive MCF7 human breast cancer cells. Note the presence of ZsGreen-positive cells in tumors, lymphatic vessels (LV), and inguinal and axillary lymph nodes (LN).

Metastatic lung tumor expressing ZsGreen1.

Figure 4. Metastatic lung tumor expressing ZsGreen1. Mouse lung bearing an MCF7+ ZsGreen tumor was fixed, embedded, and sectioned. Sections were stained with hematoxylin and eosin (left; H & E) or deparaffinized, coverslipped, and imaged via fluorescence microscopy (right) [480 nm excitation; 505 nm emission; 20X objective]. ZsGreen1 fluorescence was maintained even after histological processing.

References

  1. Stewart, N. (2006) Trends Biotechnol. 24(4):155–162.
  2. Lin, Y. et al. (2003) BMC Plant Biol. 3:10.
  3. Harrell, J. et al. (2006) Cancer Res. 66(18):9308–9315.
 

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