Name: non-invasive optical imaging of the lymphatic vasculature of a mouse
Uploaded: 2013-03-08T13:00:00
Description: Watch this Scientific Journal Video about Non-invasive Optical Imaging of the Lymphatic Vasculature of a Mouse at JoVE.com

NIH R01 CA128919 और NIH R01 HL092923: यह काम ईवा Sevick निम्नलिखित अनुदान द्वारा समर्थित किया गया.

सभी जानवरों के अध्ययन टेक्सास स्वास्थ्य विज्ञान केंद्र (ह्यूस्टन, TX), तुलनात्मक चिकित्सा विभाग, और समीक्षा और उनके संबंधित संस्थागत पशु की देखभाल द्वारा प्रोटोकॉल के अनुमोदन के बाद आण्विक इमेजिंग के ?...

<ol>
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V., McCormack, E., Mujic, M., Tenstad, O., Wiig, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimally+invasive+quantification+of+lymph+flow+in+mice+and+rats+by+imaging+depot+clearance+of+near-infrared+albumin.">Minimally invasive quantification of lymph flow in mice and rats by imaging depot clearance of near-infrared albumin.</a> Am. J. Physiol. Heart Circ. Physiol. 302, 391-401 (2012).</li><li>Zhou, Q., Wood, R., Schwarz, E. M., Wang, Y. J., Xing, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-infrared+lymphatic+imaging+demonstrates+the+dynamics+of+lymph+flow+and+lymphangiogenesis+during+the+acute+versus+chronic+phases+of+arthritis+in+mice.">Near-infrared lymphatic imaging demonstrates the dynamics of lymph flow and lymphangiogenesis during the acute versus chronic phases of arthritis in mice.</a> Arthritis Rheum. 62, 1881-1889 (2010).</li><li>Adams, K. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+evidence+of+lymphatic+function+improvement+after+advanced+pneumatic+compression+device+treatment+of+lymphedema.">Direct evidence of lymphatic function improvement after advanced pneumatic compression device treatment of lymphedema.</a> Biomed. Opt. Express. 1, 114-125 (2010).</li><li>Tan, I. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+lymphatic+contractile+function+after+manual+lymphatic+drainage+using+near-infrared+fluorescence+imaging.">Assessment of lymphatic contractile function after manual lymphatic drainage using near-infrared fluorescence imaging.</a> Arch. Phys. Med. Rehabil. 92, 756-764 (2011).</li><li>Lapinski, P. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RASA1+maintains+the+lymphatic+vasculature+in+a+quiescent+functional+state+in+mice.">RASA1 maintains the lymphatic vasculature in a quiescent functional state in mice.</a> J. Clin. Invest. 122, 733-747 (2012).</li><li>Maus, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-infrared+fluorescence+imaging+of+lymphatics+in+head+and+neck+lymphedema.">Near-infrared fluorescence imaging of lymphatics in head and neck lymphedema.</a> Head Neck. 34, 448-453 (2012).</li><li>Galanzha, E. I., Tuchin, V. V., Zharov, V. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+small+animal+mesentery+models+for+in+vivo+flow+cytometry,+dynamic+microscopy,+and+drug+screening.">Advances in small animal mesentery models for in vivo flow cytometry, dynamic microscopy, and drug screening.</a> World J. Gastroenterol. 13, 192-218 (2007).</li><li>Schramm, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cervical+lymph+node+preparation:+a+novel+approach+to+study+lymphocyte+homing+by+intravital+microscopy.">The cervical lymph node preparation: a novel approach to study lymphocyte homing by intravital microscopy.</a> Inflammation research : official journal of the European Histamine Research Society. 55, 160-167 (2006).</li><li>Hall, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+prostate+cancer+lymph+node+metastases+with+a+multimodality+contrast+agent.">Imaging prostate cancer lymph node metastases with a multimodality contrast agent.</a> Prostate. 72, 129-146 (2012).</li><li>Zhu, B., Sevick-Muraca, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimizing+excitation+leakage+and+maximizing+measurement+sensitivity+for+molecular+imaging+with+near-infrared+fluorescence.">Minimizing excitation leakage and maximizing measurement sensitivity for molecular imaging with near-infrared fluorescence.</a> J. Innovat. Opt. Health Sci. 4, 301-307 (2011).</li></ol>

हम एक रिवाज है, छोटे जानवर NIRF इमेजिंग प्रणाली का उपयोग करने के लिए लेबल चूहों में लसीका वाहिकाओं की छवियों पर कब्जा. लसीका आंदोलन की फिल्मों का निर्माण करने के लिए, 300 या अधिक छवियों को एकत्र कर रहे हैं. फि?...

<table border="1"><tbody><tr><td> समाधान, अभिकर्मकों, और उपकरण </td><td> कंपनी </td><td> कुल संख्या </td><td> टिप्पणियां </td></tr><tr><td> Indocyanine हरे (भारतीय तट रक्षक) </td><td> Patheon इटालिया एसपीए </td><td> 25431-424-02 एनडीसी </td><td> 645 सुक्ष्ममापी (5 μg/10 μL) reconstitute </td></tr><tr><td> चक्रीय Albumin बाध्यकारी डोमेन (CABD) </td><td> Bachem </td><td> रिवाज </td><td> 200 सुक्ष्ममापी (μL 6.8 μg/10) reconstitute </td></tr><tr><td> IRDye800 </td><td> ली भ्रष्टाचार </td><td> IRDye 800CW </td><td> निर्माण निर्देश के अनुसार reconstitute equilmolar सांद्रता में CABD साथ संयुग्म </td></tr><tr><td> बाँझ पानी </td><td> Hospira, इंक, झील वन, आईएल </td><td> 0409-4887-10 एनडीसी </td><td></td></tr><tr><td> नायर </td><td> चर्च और ड्वाइट कं,इंक </td><td> स्थानीय भंडार </td><td> <a href="http://www.nairlikeneverbefore.com" target="_blank">www.nairlikeneverbefore.com</a> </td></tr><tr><td> इमेजिंग प्रणाली (घटकों के नीचे) </td><td> आण्विक इमेजिंग के लिए केंद्र </td><td> N / A </td><td> हमारी प्रयोगशालाओं में कस्टम निर्मित. </td></tr><tr><td> इलेक्ट्रॉन बढ़ आरोप डिवाइस युग्मित कैमरा (EMCCD) </td><td> प्रिंसटन उपकरण, Trenton, NJ </td><td> फोटॉन 512 अधिकतम </td><td></td></tr><tr><td> Nikon कैमरा लेंस </td><td> Nikon इंक, Melville, NY </td><td> मॉडल नं 1992, Nikkor 28mm </td><td></td></tr><tr><td> ऑप्टिकल फिल्टर </td><td> Andover कार्पोरेशन, सलेम, राष्ट्रीय राजमार्ग </td><td> ANDV11333 </td><td> दो 830.0/10.0 एनएम bandpass फिल्टर लेंस के सामने में उपयोग किया जाता है </td></tr><tr><td> 785 एनएम लेजर डायोड </td><td> तीव्र लिमिटेड, North Brunswick, NJ </td><td> 1005 9mm - 78,503 </td><td> ऑप्टिकल उत्पादन की 500 मेगावाट </td></tr&gt; <tr><td> Collimating प्रकाशिकी </td><td> Thorlabs, न्यूटन, NJ </td><td> C240TME - बी </td><td> Collimates लेजर सफाई फिल्टर से पहले उत्पादन </td></tr><tr><td> क्लीन अप फिल्टर </td><td> Semrock, इंक, रोचेस्टर, NY </td><td> LD01-785/10-25 </td><td> प्रतिदीप्ति बैंड में लेजर उत्सर्जन निकालता </td></tr><tr><td> ऑप्टिकल विसारक </td><td> Thorlabs, न्यूटन, NJ </td><td> ED1-C20 </td><td> पशु से अधिक लेजर diffuses </td></tr><tr><td> वी + + </td><td> डिजिटल प्रकाशिकी, Browns बे, ऑकलैंड, न्यूजीलैंड </td><td> संस्करण 5.0 </td><td> सॉफ्टवेयर कैमरा प्रणाली को नियंत्रित करने और कंप्यूटर के लिए छवियों को बचाने के लिए इस्तेमाल किया. <a href="http://digitaloptics.net/" target="_blank">http://digitaloptics.net/</a> </td></tr><tr><td></td><td></td><td></td><td> निम्नलिखित सॉफ्टवेयर संकुल के विश्लेषणात्मक या तो सॉफ्टवेयर छवि विश्लेषण के लिए इस्तेमाल किया जा सकता है </td></tr><tr><td> ImageJ </td><td> नेशनल इंस्टीट्यूटस्वास्थ्य, Bethesda, एमडी की tutes </td><td> सबसे वर्तमान संस्करण उपलब्ध </td><td> पर फ्रीवेयर उपलब्ध <a href="http://rsbweb.nih.gov/ij/" target="_blank">http://rsbweb.nih.gov/ij/</a> </td></tr><tr><td> MATLAB </td><td> MathWorks, Natick, एमए </td><td> संस्करण 2008a या बाद में </td><td> <a href="http://www.mathworks.com/" target="_blank">http://www.mathworks.com/</a> </td></tr></tbody></table>

non-invasive optical imaging of the lymphatic vasculature of a mouse

The lymphatic vascular system is an important component of the circulatory system that maintains fluid homeostasis, provides immune surveillance, and mediates fat absorption in the gut. Yet despite its critical function, there is comparatively little understanding of how the lymphatic system adapts to serve these functions in health and disease1. Recently, we have demonstrated the ability to dynamically image lymphatic architecture and lymph "pumping" action in normal human subjects as well as in persons suffering lymphatic dysfunction using trace administration of a near-infrared fluorescent (NIRF) dye and a custom, Gen III-intensified imaging system2-4. NIRF imaging showed dramatic changes in lymphatic architecture and function with human disease. It remains unclear how these changes occur and new animal models are being developed to elucidate their genetic and molecular basis. In this protocol, we present NIRF lymphatic, small animal imaging5,6 using indocyanine green (ICG), a dye that has been used for 50 years in humans7, and a NIRF dye-labeled cyclic albumin binding domain (cABD-IRDye800) peptide that preferentially binds mouse and human albumin8. Approximately 5.5 times brighter than ICG, cABD-IRDye800 has a similar lymphatic clearance profile and can be injected in smaller doses than ICG to achieve sufficient NIRF signals for imaging8. Because both cABD-IRDye800 and ICG bind to albumin in the interstitial space8, they both may depict active protein transport into and within the lymphatics. Intradermal (ID) injections (5-50 &mu;l) of ICG (645 &mu;M) or cABD-IRDye800 (200 &mu;M) in saline are administered to the dorsal aspect of each hind paw and/or the left and right side of the base of the tail of an isoflurane-anesthetized mouse. The resulting dye concentration in the animal is 83-1,250 &mu;g/kg for ICG or 113-1,700 &mu;g/kg for cABD-IRDye800. Immediately following injections, functional lymphatic imaging is conducted for up to 1 hr using a customized, small animal NIRF imaging system. Whole animal spatial resolution can depict fluorescent lymphatic vessels of 100 microns or less, and images of structures up to 3 cm in depth can be acquired9. Images are acquired using V++ software and analyzed using ImageJ or MATLAB software. During analysis, consecutive regions of interest (ROIs) encompassing the entire vessel diameter are drawn along a given lymph vessel. The dimensions for each ROI are kept constant for a given vessel and NIRF intensity is measured for each ROI to quantitatively assess "packets" of lymph moving through vessels.

चूहों में लसीका NIRF इमेजिंग के उदाहरण जब भारतीय तट रक्षक या CABD - IRDye800 आईडी एक सामान्य माउस की पूंछ के आधार पर इंजेक्ट किया जाता है, पूंछ के आधार पर इंजेक्शन साइट और वंक्षण लिम्फ नोड (LN) के बीच लसीका वा...

Watch this Scientific Journal Video about Non-invasive Optical Imaging of the Lymphatic Vasculature of a Mouse at JoVE.com

Non-invasive Optical Imaging of the Lymphatic Vasculature of a Mouse

हाल ही में विकसित इमेजिंग निकट अवरक्त प्रतिदीप्ति (NIRF) तकनीक का उपयोग कर भूमिका लसीका प्रणाली कैंसर मेटास्टेसिस में खेलता है, प्रतिरक्षा प्रतिक्रिया, घाव की मरम्मत, और अन्य लसीका जुड़े रोगों स्पष्ट करने में मदद कर सकते हैं.

गैर इनवेसिव एक माउस के लसीका वाहिका संरचना के ऑप्टिकल इमेजिंग

The lymphatic  vascular system is an important component of the circulatory system that  maintains fluid homeostasis, provides immune surveillance, and ...

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