Name: rejection of fluorescence background in resonance and spontaneous raman microspectroscopy
Uploaded: 2011-05-18T12:00:00
Description: Watch this Scientific Journal Video about Rejection of Fluorescence Background in Resonance and Spontaneous Raman Microspectroscopy at JoVE.com

这项工作是由美国国家科学基金会奖DBI的0852891。号PHY0120999根据合作协议，由加利福尼亚大学戴维斯分校生物光子科学与技术，一个指定的美国国家科学基金会科学技术中心管理中心还资助了这项工作的一部分。

1。在准备，并把这个系统内的拉曼样品，必须采取一定的照顾。 <ol><li>由于该系统通常使用非常高数​​值孔径物镜，工作距离很短，样品置于盖玻片上。生物样品通常是放置在1号安装盖玻片厚度Attofluor细胞室（Invitrogen公司，卡尔斯巴德，加利福尼亚州）。 </li><li>液体样品，特别是那些对人体无毒，被放置在一个小玻璃瓶，巩固有机硅环氧开幕盖玻片。瓶子，然后倒测量。 </li><li>因为我们?...

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拉曼光谱生物医学领域已经看到在过去的几年中，作为其表现出的潜力解决一些困难的挑战，在生物诊断的越来越大的兴趣。例如，拉曼光谱已经证明，在癌症的检测3，4，5，6诊断价值。拉曼光谱技术也已用于细菌定量7，8和细菌对药物的反应9。它也发现在其他医学领域的应用范围从骨骼健康10 biofluid分析，11，12的广泛应用。 然而，...

<table border="1"><tbody><tr><th>名称</th><th>公司</th><th>产品编号</th><th>评论</th></tr><tr><td>镜头</td><td> ThorLabs </td><td>各种</td><td>所有的镜头涂以1％的最大传输损失</td></tr><tr><td>可调钛Sapph激光</td><td>连贯</td><td>变色龙</td><td>新泽西州，30日为200 fs，80 MHz的</td></tr><tr><td> 40X油浸泡目标</td><td>奥林巴斯</td><td> UApo/340 </td><td> NA = 1.35 </td></tr><tr><td>倒置显微镜</td><td>奥林巴斯</td><td> IX - 71 </td><td>修改，删除所有的镜头在侧面端口</td></tr><tr><td>半波片</td><td> Thorlabs </td><td> AHWP05M - 600 </td><td></td></tr><tr><td>格兰 - 汤普森偏振片</td><td> Thorlabs </td><td> GTH10M </td><td> 〜10％的传输损耗</td></tr><tr><td>光谱仪</td><td>有价证券投资阿克顿</td><td> SP2300i </td><td></td></tr><tr><td>防治荒漠化公约</td><td>有价证券投资阿克顿</td><td> PIXIS 100B </td><td></td></tr><tr><td> Mathmatical软件</td><td> The MathWorks公司</td><td> MATLAB </td><td>版本2008A </td></tr><tr><td>法拉第隔离器</td><td> EOT </td><td> BB8 - 5I </td><td></td></tr><tr><td>压电电镜</td><td>纽波特</td><td> AG - M100 </td><td></td></tr><tr><td> BBO晶体</td><td> CASIX </td><td>自定义</td><td> 1毫米的厚度</td></tr><tr><td>带通滤波器</td><td>安多弗</td><td> 008FC14 </td><td> 808 ± 0.4 nm的</td></tr><tr><td>分色镜</td><td> Semrock </td><td> FF662 - FDI01 </td><td>在662纳米带边</td></tr><tr><td>长通滤波器</td><td> Semrock </td><td> BLP01 - 405R </td><td>在417纳米带边</td></tr><tr><td>带通滤波器2 </td><td> Semrock </td><td> FF02-447/60 </td><td> 417-447纳米</td></tr><tr><td> CS 2 </td><td> Sigma - Aldrich公司</td><td> 335266 </td><td>纯度达99％ </td></tr><tr><td>香豆素30 </td><td> Sigma - Aldrich公司</td><td> 546127 </td><td>纯度达99％ </td></tr><tr><td>浸油</td><td> Cargille </td><td> 16242 </td><td> DF型</td></tr></tbody></table>

rejection of fluorescence background in resonance and spontaneous raman microspectroscopy

拉曼光谱仪是常常困扰着一个强烈的荧光背景，特别是对生物样品。如果样品是兴奋的超短脉冲，一个系统，可以暂时分离频谱重叠信号皮秒时间尺度可以隔离的火车，及时到达迟到荧光光的拉曼散射光。在这里，我们讨论一个复杂的非线性光学系统，采用全光开关在一个低功率光学克尔门的形式，隔离拉曼光谱和荧光信号的建设和运营。一个单一的808 nm的激光2.4 W的平均功率和80 MHz的重复率是分裂的，与约200毫瓦808纳米的光被转换为&lt;5兆瓦404 nm的光发送到的样本，以激发拉曼散射。其余未转换的808纳米的光，然后发送到非线性介质泵全光快门行为。快门打开和关闭了约5％的峰值效率在800 FS。使用这个系统，我们能够成功地分离拉曼光谱和荧光信号，在80 MHz的重复使用的脉冲能量和平均的权力，保持生物安全。由于该系统具有光功率没有余力，我们几个细节设计和调整的考虑，有助于最大限度地提高系统的吞吐量。我们还讨论了克尔介质内的信号和泵光束的空间和时间上的重叠，以及详细的协议光谱采集获得的协议。最后，我们报告了几个有代表性的拉曼光谱中存在强荧光，用我们的时间浇注系统取得的成果。

Watch this Scientific Journal Video about Rejection of Fluorescence Background in Resonance and Spontaneous Raman Microspectroscopy at JoVE.com

Rejection of Fluorescence Background in Resonance and Spontaneous Raman Microspectroscopy

我们讨论一个复杂的非线性光学系统，采用超快全光开关，隔离荧光信号的拉曼的建设和运营。使用这个系统，我们能够成功地分离的拉曼光谱和荧光信号，利用脉冲能量和平均的权力，保持生物安全。

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