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Imaging In Mice With Fluorescent Proteins: From Macro To Subcellular

View Article: PubMed Central - PubMed

ABSTRACT

Whole-body imaging with fluorescent proteins has been shown to be a powerful technology with many applications in small animals. Brighter, red-shifted proteins can make whole-body imaging even more sensitive due to reduced absorption by tissues and less scatter. For example, a new protein called Katushka has been isolated that is the brightest known protein with emission at wavelengths longer than 620 nm. This new protein offers potential for noninvasive whole-body macro imaging such as of tumor growth. For subcellular imaging, to observe cytoplasmic and nuclear dynamics in the living mouse, cancer cells were labeled in the nucleus with green fluorescent protein and with red fluorescent protein in the cytoplasm. The nuclear and cytoplasmic behavior of cancer cells in real time in blood vessels was imaged as they trafficked by various means or adhered to the vessel surface in the abdominal skin flap. During extravasation, real-time dual-color imaging showed that cytoplasmic processes of the cancer cells exited the vessels first, with nuclei following along the cytoplasmic projections. Both cytoplasm and nuclei underwent deformation during extravasation. Cancer cells trafficking in lymphatic vessels was also imaged. To noninvasively image cancer cell/stromal cell interaction in the tumor microenvironment as well as drug response at the cellular level in live animals in real time, we developed a new imageable three-color animal model. The model consists of GFP-expressing mice transplanted with the dual-color cancer cells. With the dual-color cancer cells and a highly sensitive small animal imaging system, subcellular dynamics can now be observed in live mice in real time. Fluorescent proteins thus enable both macro and micro imaging technology and thereby provide the basis for the new field of in vivo cell biology.

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Related in: MedlinePlus

Imaging nuclear and cytoplasmic deformation of cancer cells in the vessels in the skinA, Nondeformed cells are within a microvessel. The cells in the microvessel are round and the nuclei oval. The cells occupy the full diameter of the vessel. B, The cells and nuclei are elongated to fit a capillary. C, The cells are arrested at the capillary bifurcation. Because of the difference of the deformability between cytoplasm and nucleus, only the cytoplasm is bifurcated. The nucleus is also deformed but remains intact. D, Cytoplasmic fragmentation in very thin capillary. Bar = 50 μm [27].
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f6-sensors-08-01157: Imaging nuclear and cytoplasmic deformation of cancer cells in the vessels in the skinA, Nondeformed cells are within a microvessel. The cells in the microvessel are round and the nuclei oval. The cells occupy the full diameter of the vessel. B, The cells and nuclei are elongated to fit a capillary. C, The cells are arrested at the capillary bifurcation. Because of the difference of the deformability between cytoplasm and nucleus, only the cytoplasm is bifurcated. The nucleus is also deformed but remains intact. D, Cytoplasmic fragmentation in very thin capillary. Bar = 50 μm [27].

Mentions: With a color CCD camera, we could observe highly elongated cancer cells and nuclei in capillaries in the skin flap in living mice (Figure 6). The migration velocities of the cancer cells in the capillaries were measured by capturing images of the dual-color fluorescent cells over time. The cells and nuclei in the capillaries elongated to fit the width of these vessels. The average length of the major axis of the cancer cells in the capillaries increased to approximately four times their normal length. The nuclei increased their length 1.6 times in the capillaries. Cancer cells in capillaries over 8 μm in diameter could migrate up to 48.3 μm/hour. The data suggests that the minimum diameter of capillaries where cancer cells are able to migrate is approximately 8 μm. The use of the dual-color cancer cells differentially labeled in the cytoplasm and nucleus and associated fluorescent imaging provide a powerful tool to understand the mechanism of cancer cell migration and deformation in small vessels [27].


Imaging In Mice With Fluorescent Proteins: From Macro To Subcellular
Imaging nuclear and cytoplasmic deformation of cancer cells in the vessels in the skinA, Nondeformed cells are within a microvessel. The cells in the microvessel are round and the nuclei oval. The cells occupy the full diameter of the vessel. B, The cells and nuclei are elongated to fit a capillary. C, The cells are arrested at the capillary bifurcation. Because of the difference of the deformability between cytoplasm and nucleus, only the cytoplasm is bifurcated. The nucleus is also deformed but remains intact. D, Cytoplasmic fragmentation in very thin capillary. Bar = 50 μm [27].
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3927509&req=5

f6-sensors-08-01157: Imaging nuclear and cytoplasmic deformation of cancer cells in the vessels in the skinA, Nondeformed cells are within a microvessel. The cells in the microvessel are round and the nuclei oval. The cells occupy the full diameter of the vessel. B, The cells and nuclei are elongated to fit a capillary. C, The cells are arrested at the capillary bifurcation. Because of the difference of the deformability between cytoplasm and nucleus, only the cytoplasm is bifurcated. The nucleus is also deformed but remains intact. D, Cytoplasmic fragmentation in very thin capillary. Bar = 50 μm [27].
Mentions: With a color CCD camera, we could observe highly elongated cancer cells and nuclei in capillaries in the skin flap in living mice (Figure 6). The migration velocities of the cancer cells in the capillaries were measured by capturing images of the dual-color fluorescent cells over time. The cells and nuclei in the capillaries elongated to fit the width of these vessels. The average length of the major axis of the cancer cells in the capillaries increased to approximately four times their normal length. The nuclei increased their length 1.6 times in the capillaries. Cancer cells in capillaries over 8 μm in diameter could migrate up to 48.3 μm/hour. The data suggests that the minimum diameter of capillaries where cancer cells are able to migrate is approximately 8 μm. The use of the dual-color cancer cells differentially labeled in the cytoplasm and nucleus and associated fluorescent imaging provide a powerful tool to understand the mechanism of cancer cell migration and deformation in small vessels [27].

View Article: PubMed Central - PubMed

ABSTRACT

Whole-body imaging with fluorescent proteins has been shown to be a powerful technology with many applications in small animals. Brighter, red-shifted proteins can make whole-body imaging even more sensitive due to reduced absorption by tissues and less scatter. For example, a new protein called Katushka has been isolated that is the brightest known protein with emission at wavelengths longer than 620 nm. This new protein offers potential for noninvasive whole-body macro imaging such as of tumor growth. For subcellular imaging, to observe cytoplasmic and nuclear dynamics in the living mouse, cancer cells were labeled in the nucleus with green fluorescent protein and with red fluorescent protein in the cytoplasm. The nuclear and cytoplasmic behavior of cancer cells in real time in blood vessels was imaged as they trafficked by various means or adhered to the vessel surface in the abdominal skin flap. During extravasation, real-time dual-color imaging showed that cytoplasmic processes of the cancer cells exited the vessels first, with nuclei following along the cytoplasmic projections. Both cytoplasm and nuclei underwent deformation during extravasation. Cancer cells trafficking in lymphatic vessels was also imaged. To noninvasively image cancer cell/stromal cell interaction in the tumor microenvironment as well as drug response at the cellular level in live animals in real time, we developed a new imageable three-color animal model. The model consists of GFP-expressing mice transplanted with the dual-color cancer cells. With the dual-color cancer cells and a highly sensitive small animal imaging system, subcellular dynamics can now be observed in live mice in real time. Fluorescent proteins thus enable both macro and micro imaging technology and thereby provide the basis for the new field of in vivo cell biology.

No MeSH data available.


Related in: MedlinePlus