Elevated-temperature-induced acceleration of PACT clearing process of mouse brain tissue.
Elevated-temperature-induced acceleration of PACT clearing process of mouse brain tissue.
复制标题
升高温度引起的小鼠脑组织条约清除过程的加速度。
DOI：
10.1038/srep38848
复制
发表时间：
2017-01-31
影响因子：
4.6
通讯作者：
Zhu D
中科院分区：
文献类型：
--
作者：
Yu T;Qi Y;Zhu J;Xu J;Gong H;Luo Q;Zhu D
组织光学清除技术在高分辨率神经成像方面显示出巨大的潜力，特别是在脑内的连接方面。被动透明技术(PACT)是一种基于孵育的相对简单的透明方法，在组织透明性、荧光保存性和成像组织块的免疫染色兼容性方面具有很大的优势。然而，这种方法存在处理时间长的问题。以往的研究表明，提高温度可以加快清除速度。在这项工作中，我们的目标是系统地、定量地研究这种影响，这种影响是基于逐步升温的PACT。我们研究了不同温度下脑组织块的光学清除过程，发现温度的升高可以加快清除过程，并对荧光强度产生影响。通过权衡优缺点，我们认为42-47 °C是PACT的替代温度范围，它不仅可以产生更快的清除过程，而且可以很好地保存内源荧光，实现良好的形态保持和免疫染色兼容性，保留了PACT原有的优点。
Tissue optical clearing technique shows a great potential for neural imaging with high resolution, especially for connectomics in brain. The passive clarity technique (PACT) is a relative simple clearing method based on incubation, which has a great advantage on tissue transparency, fluorescence preservation and immunostaining compatibility for imaging tissue blocks. However, this method suffers from long processing time. Previous studies indicated that increasing temperature can speed up the clearing. In this work, we aim to systematacially and quantitatively study this influence based on PACT with graded increase of temperatures. We investigated the process of optical clearing of brain tissue block at different temperatures, and found that elevated temperature could accelerate the clearing process and also had influence on the fluorescence intensity. By balancing the advantages with drawbacks, we conclude that 42–47 °C is an alternative temperature range for PACT, which can not only produce faster clearing process, but also retain the original advantages of PACT by preserving endogenous fluorescence well, achieving fine morphology maintenance and immunostaining compatibility.