Using Ab-Space to Remove Background Components from Images in Systems of Multiple Fluorophores
Using Ab-Space to Remove Background Components from Images in Systems of Multiple Fluorophores
复制标题
使用 Ab-Space 从多荧光团系统中的图像中去除背景成分
DOI：
10.1016/j.bpj.2009.12.2138
复制
发表时间：
2010
影响因子：
3.4
通讯作者：
Clayton A
中科院分区：
文献类型：
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作者：
Clayton A
396a 2010年2月22日星期一任务。由于供体、供体-受体复合物和荧光背景的存在，生物系统通常由多个组分组成。允许从终身成像数据中去除第三个组件的工具将代表着重大进步。我们将描述一种简化的处理方法来解决二元混合物和一种新的方法来解决三元和更高的混合物使用频域程序。对于二元和三元混合物，不需要单指数衰减，这意味着每个组分可以代表多荧光团混合物。对于生物学中的许多应用，来自供体和供体-受体组分的分数荧光贡献的分辨率是可取的，因为这允许观察激活和相关参数。从频域数据来看，这可以通过使用ab坐标系的线性混合特性以一种直接的方式完成，而无需计算寿命。我们提出的理论和演示方法的解决方案，并将其应用到一个简单的生物系统。使用该技术可以很好地工作三元混合物，然而，附加频率的优势是有限的。我们已经将形式主义应用于用量子点（QDs）标记的A431细胞，量子点（QDs）有三个组成部分：QDs，细胞自身荧光和平板背景。该方法允许我们剥离自荧光和平板背景图像，只留下量子点。
396a Monday, February 22, 2010 tasks. Biological systems often consist of multiple components due to the a presence of Donor, Donor-Acceptor complex, and fluorescent background. Tools allowing a third component to be removed from lifetime imaging data would represent a significant advance. We will describe a simplified treatment for resolving binary mixtures and a novel approach to ternary and higher mixtures using frequency domain procedures. For binary and ternary mixtures there is no requirement for single exponential decay, meaning that each component can represent a multi-fluorophore mix. For many applications in biology, resolution of the fractional fluorescence contributions from the donor and donor-acceptor components is desirable as this allows activation and related parameters to be observed. From frequency domain data, this can be done in a straightforward fashion without computing lifetimes by using the linear mixing characteristics of the AB-coordinate system. We present the theory and demonstrate the approach using solutions and apply it to a simple biological system. Ternary mixtures work well using the technique, however, the advantages of additional frequencies is limited. We have applied the formalism to A431 cells labelled with quantum dots (QDs) which have three components: QDs, cellular autofluorescence and plate background. The method allowed us to strip the autofluoresence and plate background image leaving only the QDs.