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Application of GFP imaging in cancer.

Hoffman RM - Lab. Invest. (2015)

Bottom Line: Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals.Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases.With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

View Article: PubMed Central - PubMed

Affiliation: AntiCancer, Inc., Department of Surgery, University of California San Diego, San Diego, CA, USA.

ABSTRACT
Multicolored proteins have allowed the color-coding of cancer cells growing in vivo and enabled the distinction of host from tumor with single-cell resolution. Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals. Non-invasive imaging of cancer cells expressing fluorescent proteins has allowed the real-time determination of efficacy of candidate antitumor and antimetastatic agents in mouse models. The use of fluorescent proteins to differentially label cancer cells in the nucleus and cytoplasm can visualize the nuclear-cytoplasmic dynamics of cancer cells in vivo including: mitosis, apoptosis, cell-cycle position, and differential behavior of nucleus and cytoplasm that occurs during cancer-cell deformation and extravasation. Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases. With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

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an example of the earliest prototype for in vivo imaging with GFP is the Illumatool a simple instrument with a light sources that are properly filtered to avoid autofluorescence and an emission filter through which it is possible to image GFP fluorescence from unrestrained animals.11 An example of the best state of the art, OV100 small animal imaging system: The OV-100 Small Animal Imaging System (Olympus, Tokyo, Japan), containing an MT-20 light source (Olympus) and DP70 CCD camera (Olympus) was used. The optics of the OV-100 fluorescence imaging system have been specially developed for macroimaging as well as microimaging with high light-gathering capacity. The instrument incorporates a unique combination of high numerical aperture and long working distance. Five individually optimized objective lenses, parcentered and parfocal, provide a 105-fold magnification range for seamless imaging of the entire body down to the subcellular level without disturbing the animal. The OV-100 has the lenses mounted on an automated turret with a high magnification range of ×1.6 to ×16 and a field of view ranging from 6.9 to 0.69 mm. The optics and antireflective coatings ensure optimal imaging of multiplexed fluorescent reporters in small animals. High-resolution images were captured directly on a PC (Fujitsu Siemens, Munich, Germany). Images were processed for contrast and brightness and analyzed with the use of Paint Shop Pro 8 and CellR (Olympus Biosystems).13
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Figure 1: an example of the earliest prototype for in vivo imaging with GFP is the Illumatool a simple instrument with a light sources that are properly filtered to avoid autofluorescence and an emission filter through which it is possible to image GFP fluorescence from unrestrained animals.11 An example of the best state of the art, OV100 small animal imaging system: The OV-100 Small Animal Imaging System (Olympus, Tokyo, Japan), containing an MT-20 light source (Olympus) and DP70 CCD camera (Olympus) was used. The optics of the OV-100 fluorescence imaging system have been specially developed for macroimaging as well as microimaging with high light-gathering capacity. The instrument incorporates a unique combination of high numerical aperture and long working distance. Five individually optimized objective lenses, parcentered and parfocal, provide a 105-fold magnification range for seamless imaging of the entire body down to the subcellular level without disturbing the animal. The OV-100 has the lenses mounted on an automated turret with a high magnification range of ×1.6 to ×16 and a field of view ranging from 6.9 to 0.69 mm. The optics and antireflective coatings ensure optimal imaging of multiplexed fluorescent reporters in small animals. High-resolution images were captured directly on a PC (Fujitsu Siemens, Munich, Germany). Images were processed for contrast and brightness and analyzed with the use of Paint Shop Pro 8 and CellR (Olympus Biosystems).13

Mentions: A variable-magnification small animals imaging system (OV100, Olympus Corp., Tokyo, Japan), containing an MT-20 light source (Olympus Biosystems, Planegg, Germany) and DP70 CCD camera (Olympus), can be used for macro and subcellular imaging in live mice. The optics of the OV100 fluorescence imaging system have been specially developed for macroimaging as well as microimaging with high light-gathering capacity. The objectives have high numerical aperture and are long working distance. Individually optimized objective lenses, parcentered and parfocal, provide a 105-fold magnification range for imaging of the entire body down to the subcellular level without disturbing the animal. The OV100 has the lenses mounted on an automated turret with a high magnification range of ×1.6 to ×16 and a field of view ranging from 6.9 to 0.69 mm. The optics and anti-reflective coatings ensure optimal imaging of multiplexed fluorescent reporters in small animals (Figure 1).13


Application of GFP imaging in cancer.

Hoffman RM - Lab. Invest. (2015)

an example of the earliest prototype for in vivo imaging with GFP is the Illumatool a simple instrument with a light sources that are properly filtered to avoid autofluorescence and an emission filter through which it is possible to image GFP fluorescence from unrestrained animals.11 An example of the best state of the art, OV100 small animal imaging system: The OV-100 Small Animal Imaging System (Olympus, Tokyo, Japan), containing an MT-20 light source (Olympus) and DP70 CCD camera (Olympus) was used. The optics of the OV-100 fluorescence imaging system have been specially developed for macroimaging as well as microimaging with high light-gathering capacity. The instrument incorporates a unique combination of high numerical aperture and long working distance. Five individually optimized objective lenses, parcentered and parfocal, provide a 105-fold magnification range for seamless imaging of the entire body down to the subcellular level without disturbing the animal. The OV-100 has the lenses mounted on an automated turret with a high magnification range of ×1.6 to ×16 and a field of view ranging from 6.9 to 0.69 mm. The optics and antireflective coatings ensure optimal imaging of multiplexed fluorescent reporters in small animals. High-resolution images were captured directly on a PC (Fujitsu Siemens, Munich, Germany). Images were processed for contrast and brightness and analyzed with the use of Paint Shop Pro 8 and CellR (Olympus Biosystems).13
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383682&req=5

Figure 1: an example of the earliest prototype for in vivo imaging with GFP is the Illumatool a simple instrument with a light sources that are properly filtered to avoid autofluorescence and an emission filter through which it is possible to image GFP fluorescence from unrestrained animals.11 An example of the best state of the art, OV100 small animal imaging system: The OV-100 Small Animal Imaging System (Olympus, Tokyo, Japan), containing an MT-20 light source (Olympus) and DP70 CCD camera (Olympus) was used. The optics of the OV-100 fluorescence imaging system have been specially developed for macroimaging as well as microimaging with high light-gathering capacity. The instrument incorporates a unique combination of high numerical aperture and long working distance. Five individually optimized objective lenses, parcentered and parfocal, provide a 105-fold magnification range for seamless imaging of the entire body down to the subcellular level without disturbing the animal. The OV-100 has the lenses mounted on an automated turret with a high magnification range of ×1.6 to ×16 and a field of view ranging from 6.9 to 0.69 mm. The optics and antireflective coatings ensure optimal imaging of multiplexed fluorescent reporters in small animals. High-resolution images were captured directly on a PC (Fujitsu Siemens, Munich, Germany). Images were processed for contrast and brightness and analyzed with the use of Paint Shop Pro 8 and CellR (Olympus Biosystems).13
Mentions: A variable-magnification small animals imaging system (OV100, Olympus Corp., Tokyo, Japan), containing an MT-20 light source (Olympus Biosystems, Planegg, Germany) and DP70 CCD camera (Olympus), can be used for macro and subcellular imaging in live mice. The optics of the OV100 fluorescence imaging system have been specially developed for macroimaging as well as microimaging with high light-gathering capacity. The objectives have high numerical aperture and are long working distance. Individually optimized objective lenses, parcentered and parfocal, provide a 105-fold magnification range for imaging of the entire body down to the subcellular level without disturbing the animal. The OV100 has the lenses mounted on an automated turret with a high magnification range of ×1.6 to ×16 and a field of view ranging from 6.9 to 0.69 mm. The optics and anti-reflective coatings ensure optimal imaging of multiplexed fluorescent reporters in small animals (Figure 1).13

Bottom Line: Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals.Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases.With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

View Article: PubMed Central - PubMed

Affiliation: AntiCancer, Inc., Department of Surgery, University of California San Diego, San Diego, CA, USA.

ABSTRACT
Multicolored proteins have allowed the color-coding of cancer cells growing in vivo and enabled the distinction of host from tumor with single-cell resolution. Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals. Non-invasive imaging of cancer cells expressing fluorescent proteins has allowed the real-time determination of efficacy of candidate antitumor and antimetastatic agents in mouse models. The use of fluorescent proteins to differentially label cancer cells in the nucleus and cytoplasm can visualize the nuclear-cytoplasmic dynamics of cancer cells in vivo including: mitosis, apoptosis, cell-cycle position, and differential behavior of nucleus and cytoplasm that occurs during cancer-cell deformation and extravasation. Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases. With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

Show MeSH
Related in: MedlinePlus