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data-appid="datumInfo" data-config="" data-state="true"> <div id="4eumywqoc8" class="appMask"></div> <div id="4eumywqoc8" class="appContent"> <div id="4eumywqoc8" class="datum_info"> <h1 class="title">ELISPOT斑點的特征</h1> <div id="4eumywqoc8" class="bgcolor_01"> <div id="4eumywqoc8" class="bgcolor_sum"> <div id="4eumywqoc8" class="left">日期：2024-11-22 05:59</div> <div id="4eumywqoc8" class="right">瀏覽次數(shù)：806</div> </div> <div id="4eumywqoc8" class="bgcolor_sum01"> 摘要： “一個陽性細胞，對應一個ELISPOT斑點”，這是ELISPOT檢測的理論前提。無數(shù)的實驗也證明了這一點?，F(xiàn)有的抗體技術(shù)以及酶聯(lián)放大技術(shù)確實可以把單個細胞分泌細胞因子的行為記錄下來，ELISPOT板上的每一個斑點就是*好的證明。下面我們來看一看ELISPOT斑點的特征。圓形，有暈核結(jié)構(gòu) 這是判定真假ELISPOT斑點的主要標準。ELISPOT斑點有這種結(jié)構(gòu)可以從幾何學的原理得到很好的理解和解釋（如圖 1.2所示）。懸浮在ELISPOT板膜上的細胞，它分泌細胞因子是沒有方向... </div> </div> <div id="4eumywqoc8" class="content"> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;">“一個陽性細胞，對應一個ELISPOT斑點”，這是ELISPOT檢測的理論前提。無數(shù)的實驗也證明了這一點?，F(xiàn)有的抗體技術(shù)以及酶聯(lián)放大技術(shù)確實可以把單個細胞分泌細胞因子的行為記錄下來，ELISPOT板上的每一個斑點就是*好的證明。下面我們來看一看ELISPOT斑點的特征。</span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;"><br> </span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;"> <strong><span style="font-size:16px;">圓形，有暈核結(jié)構(gòu)</span></strong></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;"><img src="http://www.bio-city.net/uploads/common/image/20130830/20130830-100030_665055216.png" align="right" alt="" style="border-style:none;vertical-align:top;display:block;margin:0px auto;"></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;"><br> </span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;"> 這是判定真假ELISPOT斑點的主要標準。ELISPOT斑點有</span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;">這種結(jié)構(gòu)可以從幾何學的原理得到很好的理解和解釋（如圖</span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;">1.2所示）。懸浮在ELISPOT板膜上的細胞，它分泌細胞因子</span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;">是沒有方向性的，均勻地向四面八方擴散，如果擴散到</span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;">ELISPOT板膜，就會被板膜上的包被抗體捕獲。在所有從細胞到斑膜的路徑中，有條路徑*短，</span><span style="font-size:14px;color:#333333;line-height:1.5;">這就是位于細胞正下方到板膜的垂直距離。路徑越短，擴散中耗散損失就越少，到達該點的細胞</span><span style="font-size:14px;color:#333333;line-height:1.5;">因子就越多，將來顯色的時候顏色就越深。以垂直距離點為圓心，向外畫一系列的同心圓，同心</span><span style="font-size:14px;color:#333333;line-height:1.5;">圓的直徑越大，細胞因子擴散的距離就越長，耗散越多，到達的細胞因子就越少，顏色就越淺。</span><span style="font-size:14px;color:#333333;line-height:1.5;">當超過一定距離之后，擴散到達的細胞因子就減少到背景水平了，這就是斑點的邊界。</span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;"><br> </span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;"> <strong><span style="font-size:16px;"> 大小不均一，正態(tài)分布</span></strong></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;"><strong><span style="font-size:16px;"><br> </span></strong></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"> <span style="font-size:14px;color:#333333;"><img src="http://www.bio-city.net/uploads/common/image/20130830/20130830-100022_243427792.png" align="right" alt="" style="border-style:none;vertical-align:top;display:block;margin:0px auto;"></span> </p> <span style="font-size:14px;font-family:'Microsoft Yahei', arial, 宋體;white-space:normal;color:#333333;background-color:#FFFFFF;"> <p style="margin-top:0px;margin-bottom:0px;padding:0px;"> <span style="line-height:1.5;"> </span><span style="line-height:1.5;"> ELISPOT斑點要比產(chǎn)生它的那個細胞大很多。我們知道，</span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;"> <span style="line-height:1.5;">哺乳動物細胞的直徑約是5-10μm，而ELISPOT斑點的直徑通常是50-150μm，比細胞大10倍以上。</span> </p> </span><span style="font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"></span><span style="font-size:14px;font-family:'Microsoft Yahei', arial, 宋體;white-space:normal;color:#333333;background-color:#FFFFFF;"> <div style="margin:0px;padding:0px;text-align:justify;"> <span style="line-height:1.5;"> 同一類陽性細胞，受到相同的刺激，產(chǎn)生的斑點大小也不完全相同。這些斑點大小不一，但是如果把它們的直徑統(tǒng)計出來，這發(fā)現(xiàn)它們呈正態(tài)分布的，見圖1.3。</span> </div> </span><span style="font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"></span><span style="font-size:14px;font-family:'Microsoft Yahei', arial, 宋體;white-space:normal;color:#333333;background-color:#FFFFFF;"> <div style="margin:0px;padding:0px;text-align:justify;"> <span style="line-height:1.5;"> 需要說明的是，不同的細胞，對不同的刺激物，不同的細胞因子，ELISPOT斑點的直徑（平均直徑）也不同。通常來說，T細胞產(chǎn)生的斑點較大，單核巨噬細胞產(chǎn)生的斑點??；特異性的抗原（多肽或者蛋白）刺激產(chǎn)生的斑點大，非特異性的有絲分裂原（PHA、ConA等）刺激產(chǎn)生的斑點??；IFN-γ、IL-4等細胞因子的斑點大，IL-10、IL-12、Granzyme B之類的細胞因子的斑點小。</span> </div> </span><span style="font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;background-color:#FFFFFF;"></span><span style="font-size:14px;font-family:'Microsoft Yahei', arial, 宋體;white-space:normal;color:#333333;background-color:#FFFFFF;"> <div style="margin:0px;padding:0px;text-align:justify;"> <span style="line-height:1.5;"> 如果一次實驗中有兩類細胞同時受到刺激，比如T細胞與巨噬細胞，或者一次實驗中T細胞受到兩種不同的刺激物刺激，比如蛋白抗原與內(nèi)**（大腸桿菌表達的蛋白很可能含有較高濃度的內(nèi)**），那么同一個孔中就會出現(xiàn)兩種不同的斑點。這兩種斑點用肉眼很難區(qū)別，但是如果對它們直徑分布做統(tǒng)計作圖的話，就會有別于圖1.3中的單峰分布，而是呈現(xiàn)雙峰分布。</span> </div> </span> </div> <div id="4eumywqoc8" class="datumrelate"> <span>下一篇：</span> <a href="/klbscience_Article_6882272.html" title="為什么需要自動化的ELISPOT圖像分析儀？">為什么需要自動化的ELISPOT圖像分析儀？</a> <br> <span>上一篇：</span> <a href="/klbscience_Article_7015657.html" title="血清對ELISPOT檢測的干擾">血清對ELISPOT檢測的干擾</a> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> <footer id="footer" class="footer" 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