活细胞怎么计数？

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  烟雨笑春风 2019-04-19 17:07:04

  <p><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">用台盼蓝拒染法计数活细胞:</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">1. 彻底混合细胞悬液,取100礚样本至一支试管中.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">2. 台盼蓝染色有两种方法.一是首先用细胞培养液稀释细胞样本,然后用台盼蓝溶液(产品号 #07050) 1/2稀释样本（细胞和台盼蓝的稀释比为1:1）.或者直接用台盼蓝溶液稀释样本.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">例如：1/40稀释样本</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">加入50礚 细胞悬液至950礚 IMDM+2% FBS (1/20 稀释),混合,然后取100礚稀释的细胞悬液加入100礚台盼蓝溶液中(终稀释度为1/40).或取20礚细胞悬液加入780礚台盼蓝溶液中(终稀释度为1/40).</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">3. 涡悬振荡,混合稀释的细胞样本.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">4. 血细胞计数器的准备：首先用酒精清洁其小室和盖玻片,然后用脱脂绵擦干.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">5. 仔细的将盖玻片覆盖两个小室.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">6. 用微量移液器或毛细管吸取稀释的样本.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">7. 用微量移液器或毛细管将样本充满两个小室.不要过满或不足.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">8. 计数4个大方格中细胞核的总数(1x1x0.1mm)或&gt;100个细胞.分别死细胞和活细胞.死细胞染成蓝色的死细胞(因为细胞完整性破坏,细胞吸收台盼蓝),活细胞透明有折光（未染色）.</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">9. 活细胞计数按以下方法计算：</span><br style="content: &quot;&quot;; display: block; width: 705px; height: 0px; margin: 20px 0px; color: rgb(51, 51, 51); font-family: &quot;PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; white-space: normal; background-color: rgb(255, 255, 255);"/><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);PingFang SC&quot;, &quot;Lantinghei SC&quot;, &quot;Microsoft YaHei&quot;, arial, 宋体, sans-serif, tahoma; background-color: rgb(255, 255, 255);" data-x-flag="17_6">每个方格中活细胞总数平均值x稀释倍数x104= 活细胞数/mL</span></p>

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