Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications.
Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications.
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DOI：
10.1117/1.2967184
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发表时间：
2008-07
影响因子：
3.5
通讯作者：
Ntziachristos V
中科院分区：
文献类型：
--
作者：
Deliolanis NC;Kasmieh R;Wurdinger T;Tannous BA;Shah K;Ntziachristos V
新的荧光蛋白（FPs）的发现，在光谱的远红色部分发射，从组织吸收的光明显低于可见光，提供了在整个器官或小动物体内水平进行非侵入性生物检测的可能性。FPs在深层组织成像中的性能不仅取决于其光学特性，还取决于波长相关的组织吸收和荧光活性的深度。为了确定FP和照明波长的最佳选择，我们比较了5种最有前途的FP的性能，即tdTomato、mCherry、maspberry、mPlum和Katushka。我们通过实验测量了小鼠的信号强度，并采用理论预测来了解不同照明情况下的性能，特别是与层压成像有关的性能。结果发现，与绿色荧光蛋白（GFP）相比，适当的红移蛋白和照明波长组合可以使小动物的检测灵敏度提高至少2个数量级。血红蛋白光谱在600nm范围内的急剧衰减变化可能会显著影响检测灵敏度，因此需要仔细选择照明波长以获得最佳成像性能。
The discovery of new fluorescent proteins (FPs) that emit in the far-red part of the spectrum, where light absorption from tissue is significantly lower than in the visible, offers the possibility for non-invasive biological interrogation at the entire organ or small animal level in vivo. The performance of FPs in deep-tissue imaging depends not only on their optical characteristics, but also on the wavelength-dependent tissue absorption and the depth of the fluorescence activity. In order to determine the optimal choice of FP and illumination wavelength we compared the performance of five of the most promising FPs, i.e. tdTomato, mCherry, mRaspberry, mPlum, and Katushka. We experimentally measured the signal strength through mice and employed theoretical predictions to obtain an understanding on the performance of different illumination scenarios, especially as it pertains to tomographic imaging. It was found that the appropriate combination of red-shifted proteins and illumination wavelengths can improve detection sensitivity in small animals by at least 2 orders of magnitude compared to Green FP (GFP). It is also shown that the steep attenuation change of the hemoglobin spectrum around the 600nm range may significantly affect the detection sensitivity and necessitates the careful selection of illumination wavelengths for optimal imaging performance.
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影响因子：
3.4
作者：
Patterson, GH;Knobel, SM;Piston, DW
通讯作者：
Piston, DW
影响因子：
4.8
作者：
Winnard, Paul T., Jr.;Kluth, Jessica B.;Raman, Venu
通讯作者：
Raman, Venu
DOI：
10.1073/pnas.051626698
发表时间：
2001-02-27
影响因子：
11.1
作者：
Yang, M;Baranov, E;Hoffman, RM
通讯作者：
Hoffman, RM
DOI：
10.1073/pnas.97.3.1206
发表时间：
2000-02-01
影响因子：
11.1
作者：
Yang, M;Baranov, E;Hoffman, RM
通讯作者：
Hoffman, RM
影响因子：
2.8
作者：
Zacharakis, Giannis;Shih, Helen;Ntziachristos, Vasilis
通讯作者：
Ntziachristos, Vasilis