无论在脊椎动物还是无脊椎动物中,提高神经环路的成像效果对于神经环路的功能研究都是必不可少的。因此,一种新的基于鼠脑的成像技术——清晰化技术(CLARITY),正吸引越来越多的研究者将它运用到其它物种中。这里我们将改进过的清晰化技术应用于无脊椎动物海兔的神经节的清晰化成像中。改进之处包括为海兔神经节修改了水凝胶溶液配方并设计了一个专用的容器。我们使用预先经过免疫组化处理的神经节,再使用改进的清晰化技术。我们采用荧光和共聚焦成像检查这些技术的相容性以及成像效果的改善程度。结果显示,这种改进过的清晰化技术确实可以使样本更清晰,并且进一步提高了海兔神经节中神经肽CP2免疫阳性神经元的清晰度。例如,这种方法提高了神经节深处的一些弱免疫阳性神经元的可视化效果。这种改良方法使得清晰化技术不仅更适合海兔整体神经节的结构成像,也会对其它的无脊椎动物神经节的成像起到帮助作用。
Improvements in the imaging of neural circuits are essential for studies of network function in both invertebrates and verte- brates. Therefore, CLARITY, a new imaging enhancement technique developed for mouse brains has attracted broad interest from researchers working on other species. We studied the potential of a modified version of CLARITY to enhance the imaging of ganglia in an invertebrate Aplysia. For example, we have modified the hydrogel solution and designed a small container for the Aplysia ganglia. The ganglia were first processed for immunohistochemistry, and then for CLARITY. We examined the compatibility of these techniques and the extent to which the imaging of fluorescence improved using confocal microscopy. We found that CLARITY did indeed enhance the imaging of CP2 immunopositive neurons in Aplysia ganglia. For example, it improved visualization of small, weak immunoreactive neurons deep in the ganglia. Our modifications of CLARITY make this new method suitable for future use in Aplysia experiments. Furthermore, our techniques are likely to facilitate imaging in other invertebrate ganglia.