In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent reporters

Palmer, Gregory M.; Fontanella, Andrew N.; Shan, Siqing; Hanna, Gabi; Zhang, Guoqing; Fraser, Cassandra L.; Dewhirst, Mark W.

doi:10.1038/nprot.2011.349

Cited by 159 publications

(153 citation statements)

References 22 publications

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“…The skinfold chamber, which consists of two symmetrical titanium frames, was implanted in the dorsal skin of the mice following previously reported procedures (40)(41)(42). The chamber sandwiched and immobilized the skin on the back of the mouse.…”

Section: Dorsal Skinfold Chamber Model In Micementioning

confidence: 99%

Label-free DNA imaging in vivo with stimulated Raman scattering microscopy

Lü

Basu

Igras

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

252

213

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Label-free DNA imaging is highly desirable in biology and medicine to perform live imaging without affecting cell function and to obtain instant histological tissue examination during surgical procedures. Here we show a label-free DNA imaging method with stimulated Raman scattering (SRS) microscopy for visualization of the cell nuclei in live animals and intact fresh human tissues with subcellular resolution. Relying on the distinct Raman spectral features of the carbon-hydrogen bonds in DNA, the distribution of DNA is retrieved from the strong background of proteins and lipids by linear decomposition of SRS images at three optimally selected Raman shifts. Based on changes on DNA condensation in the nucleus, we were able to capture chromosome dynamics during cell division both in vitro and in vivo. We tracked mouse skin cell proliferation, induced by drug treatment, through in vivo counting of the mitotic rate. Furthermore, we demonstrated a label-free histology method for human skin cancer diagnosis that provides comparable results to other conventional tissue staining methods such as H&E. Our approach exhibits higher sensitivity than SRS imaging of DNA in the fingerprint spectral region. Compared with spontaneous Raman imaging of DNA, our approach is three orders of magnitude faster, allowing both chromatin dynamic studies and label-free optical histology in real time.

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Section: Dorsal Skinfold Chamber Model In Micementioning

confidence: 99%

Label-free DNA imaging in vivo with stimulated Raman scattering microscopy

Lü

Basu

Igras

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

252

213

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“…To assess the ability of the engineered muscle to survive and vascularize in vivo, we implanted 2-wk predifferentiated (PreD) bundles into a dorsal skinfold window chamber in nude mice (29). For this purpose, we generated smaller three-bundle constructs anchored within a single 9 × 9-mm square Cerex frame that fit within the window chamber (Fig.…”

Section: /Myodmentioning

confidence: 99%

Biomimetic engineered muscle with capacity for vascular integration and functional maturation in vivo

Juhas

Engelmayr

Fontanella

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

209

316

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Tissue-engineered skeletal muscle can serve as a physiological model of natural muscle and a potential therapeutic vehicle for rapid repair of severe muscle loss and injury. Here, we describe a platform for engineering and testing highly functional biomimetic muscle tissues with a resident satellite cell niche and capacity for robust myogenesis and self-regeneration in vitro. Using a mouse dorsal window implantation model and transduction with fluorescent intracellular calcium indicator, GCaMP3, we nondestructively monitored, in real time, vascular integration and the functional state of engineered muscle in vivo. During a 2-wk period, implanted engineered muscle exhibited a steady ingrowth of blood-perfused microvasculature along with an increase in amplitude of calcium transients and force of contraction. We also demonstrated superior structural organization, vascularization, and contractile function of fully differentiated vs. undifferentiated engineered muscle implants. The described in vitro and in vivo models of biomimetic engineered muscle represent enabling technology for novel studies of skeletal muscle function and regeneration.tissue engineering | contractile force | self-repair | angiogenesis | window chamber

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“…The techniques described here are also compatible with surgically placed window chambers 8,42,43 for longitudinal studies. The Procedure section consists of a step-by-step protocol for the study of nanoparticle transport and cell-particle interactions in the liver, followed by Application Notes section that describes how this protocol may be adapted for subcutaneous tumor imaging.…”

Section: Experimental Designmentioning

confidence: 94%

“…Dynamic biological processes for which quantitative IVM procedures have been implemented include the assessment of vascular morphology 8 , permeability 9 , flow 10 , and response to therapy 11 . IVM is also well suited for imaging the dynamics of nanoparticles in vivo, and as result, IVM is increasingly leveraged for the design and optimization of nanotherapeutics [12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29] .…”

Section: Introductionmentioning

confidence: 99%

Real-time intravital microscopy of individual nanoparticle dynamics in liver and tumors of live mice

Ven¹,

Yun²,

Kim³

et al. 2013

Protocol Exchange

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Intravital microscopy is emerging as an important experimental tool for the research and development of multi-functional therapeutic nanoconstructs. The direct visualization of nanoparticle dynamics within live animals provides invaluable insights into the mechanisms that regulate nanotherapeutics transport and cell-particle interactions. Here we present a protocol to image the dynamics of nanoparticles within the liver and tumors of live mice immediately following systemic injection using a high-speed (30-400 fps) confocal or multi-photon laserscanning fluorescence microscope. Techniques for quantifying the real-time accumulation and cellular association of individual particles with a size ranging from several tens of nanometers to micrometers are described, as well as an experimental strategy for labeling Kupffer cells in the liver in vivo. Experimental design considerations and controls are provided, as well as minimum equipment requirements. The entire protocol takes approximately 4-8 hours and yields quantitative information. These techniques can serve to study a wide range of kinetic parameters that drive nanotherapeutics delivery, uptake, and treatment response.

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In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent reporters

Cited by 159 publications

References 22 publications

Label-free DNA imaging in vivo with stimulated Raman scattering microscopy

Label-free DNA imaging in vivo with stimulated Raman scattering microscopy

Biomimetic engineered muscle with capacity for vascular integration and functional maturation in vivo

Real-time intravital microscopy of individual nanoparticle dynamics in liver and tumors of live mice

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