Name: sublimation of dan matrix for the detection and visualization of gangliosides in rat brain tissue for maldi imaging mass spectrometry
Uploaded: 2017-03-23T13:00:00
Description: Watch this Scientific Journal Video about Sublimation of DAN Matrix for the Detection and Visualization of Gangliosides in Rat Brain Tissue for MALDI Imaging Mass Spectrometry at JoVE.com

We would like to acknowledge the technical assistance of Kristina Jurcic in the University of Western Ontario MALDI MS facility, as well as the National Sciences and Engineering Council (NSERC) for funding this work. The authors would also like to acknowledge the Caprioli group (Vanderbilt University, TN) and Chaurand group (Universit&#233; de Montreal, QC) for their advice in optimizing the sublimation technique presented in this manuscript.

सभी पशु हैंडलिंग प्रक्रियाओं वेस्टर्न ओंटारियो के जानवरों की देखभाल समिति (2016-014) विश्वविद्यालय का पालन नीचे वर्णित है। 1. ऊतक तैयारी और सेक्शनिंग <ol><li> के रूप में &quot;ताजा जमे हुए निष्कर्षण&quot; (य?...

<ol>
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यह काम एक मानकीकृत ऐसे gangliosides के रूप में नकारात्मक आरोप लगाया लिपिड, MALDI आईएमएस प्रयोगों में, का पता लगाने के लिए ऊतक पर मैट्रिक्स बनाने की क्रिया के प्रोटोकॉल का विवरण। MALDI आईएमएस के लिए नमूना तैयार अत्यधि?...

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Sublimator</td>
			<td>Chemglass Life Sciences</td>
			<td>CG3038-01</td>
			<td></td>
		</tr>
		<tr>
			<td>1,5- Diaminonapthalene (DAN) matrix</td>
			<td>Sigma-Aldrich</td>
			<td>D21200&#160;</td>
			<td>100 G</td>
		</tr>
		<tr>
			<td>Cryostat</td>
			<td>Thermo-Fisher Scientific</td>
			<td></td>
			<td>CryoStar NX50</td>
		</tr>
		<tr>
			<td>Hot plate with temperature feedback</td>
			<td>Thermo-Fisher Scientific</td>
			<td>HP88857290</td>
			<td>Isotemp ADVD 7x7 HP 100-120v</td>
		</tr>
		<tr>
			<td>Stainless Steel Jack</td>
			<td>Thermo-Fisher Scientific</td>
			<td>2216479</td>
			<td>10x10</td>
		</tr>
		<tr>
			<td>Cold Trap</td>
			<td></td>
			<td></td>
			<td>Custom built on site</td>
		</tr>
		<tr>
			<td>Vacuum Pump</td>
			<td>Franklin Electric</td>
			<td>1102180403</td>
			<td>Savant VP100 Two Stage</td>
		</tr>
		<tr>
			<td>Indium-tin-oxide (ITO) Slides</td>
			<td>Hudson Surface Technology</td>
			<td>PSI 1111000</td>
			<td>type II, 1.1mm/25 each</td>
		</tr>
		<tr>
			<td>MALDI TOF/TOF 5800 Instrument</td>
			<td>AB Sciex</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Desiccator</td>
			<td>Sigma-Aldrich</td>
			<td>D2797</td>
			<td>tabletop desiccator</td>
		</tr>
	</tbody>
</table>

sublimation of dan matrix for the detection and visualization of gangliosides in rat brain tissue for maldi imaging mass spectrometry

नमूना तैयार इष्टतम का पता लगाने और मैट्रिक्स की मदद से desorption लेजर / आयनीकरण (MALDI) इमेजिंग मास स्पेक्ट्रोमेट्री (आईएमएस) प्रयोगों में analytes के दृश्य के लिए महत्वपूर्ण है। उचित प्रोटोकॉल का निर्धारण नमूना तैयार करने की प्रक्रिया के दौरान पालन करने के लिए मुश्किल हो सकता है के रूप में हर कदम ब्याज की analytes की अद्वितीय विशेषताओं के साथ पालन करने के लिए अनुकूलित किया जाना चाहिए। यह प्रक्रिया केवल एक संगत मैट्रिक्स है कि desorb और कुशलता से ब्याज के अणुओं योण सकते हैं खोजने नहीं शामिल है, लेकिन यह भी उचित मैट्रिक्स बयान तकनीक का चयन। उदाहरण के लिए, एक गीला मैट्रिक्स बयान तकनीक है, जो विलायक में एक मैट्रिक्स भंग जरूरत पर जोर देता है, सबसे अधिक प्रोटीन और पेप्टाइड्स के desorption के लिए बेहतर है, जबकि सूखी मैट्रिक्स बयान तकनीक लिपिड के आयनीकरण लिए विशेष रूप से प्रभावी हैं। उच्च बनाने की क्रिया homogenei के कारण MALDI आईएमएस द्वारा ऊतकों में लिपिड का पता लगाने के लिए सूखी मैट्रिक्स बयान के एक अत्यंत कुशल पद्धति के रूप में सूचित किया गया हैमैट्रिक्स क्रिस्टल बयान और कम से कम analyte delocalization की Ty कई गीला बयान तरीकों 1, 2 की तुलना में। मोटे तौर पर, यह नमूने एक ठंडी सतह के खिलाफ दबाया के साथ एक निर्वात सील कक्ष में एक नमूना और पाउडर मैट्रिक्स रखने शामिल है। तंत्र तो, एक गर्म स्नान (रेत या तेल) में उतारा कूल्ड ऊतक का नमूना की सतह पर पाउडर मैट्रिक्स के उच्च बनाने की क्रिया में जिसके परिणामस्वरूप है। यहाँ हम पता लगाने और MALDI आईएमएस का उपयोग कर चूहे के मस्तिष्क में gangliosides के दृश्य के लिए 1,5-diaminonaphthalene (दान) मैट्रिक्स का उपयोग कर एक उच्च बनाने की क्रिया प्रोटोकॉल का वर्णन।

उच्च बनाने की क्रिया प्रयोग के पूरा होने पर, कांच के उपकरण के दो हिस्सों, धूआं डाकू और स्लाइड में अलग हो रहे हैं कंडेनसर के लिए टेप, हटाया जा सकता है (चित्रा 3 ए)। इस बिंदु पर, स्लाइड असमान...

Watch this Scientific Journal Video about Sublimation of DAN Matrix for the Detection and Visualization of Gangliosides in Rat Brain Tissue for MALDI Imaging Mass Spectrometry at JoVE.com

Sublimation of DAN Matrix for the Detection and Visualization of Gangliosides in Rat Brain Tissue for MALDI Imaging Mass Spectrometry

A protocol for the sublimation of DAN matrix onto rat brain tissue for the detection of gangliosides using MALDI Imaging Mass Spectrometry is presented.

MALDI इमेजिंग मास स्पेक्ट्रोमेट्री के लिए चूहा मस्तिष्क के ऊतकों में जांच और gangliosides के दृश्य के लिए दान मैट्रिक्स की उच्च बनाने की क्रिया

Sample preparation is key for optimal detection and visualization of analytes in Matrix-assisted Laser Desorption/Ionization (MALDI) Imaging Mass ...

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