Peterson Brittni M, Mermelstein Paul G, Meisel Robert L
Graduate Program in Neuroscience, University of Minnesota, 6-145 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455, USA; Department of Neuroscience, University of Minnesota, 6-145 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455, USA.
Graduate Program in Neuroscience, University of Minnesota, 6-145 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455, USA; Department of Neuroscience, University of Minnesota, 6-145 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455, USA.
J Neurosci Methods. 2015 Mar 15;242:106-11. doi: 10.1016/j.jneumeth.2015.01.014. Epub 2015 Jan 17.
Structural plasticity, such as changes in dendritic spine morphology and density, reflect changes in synaptic connectivity and circuitry. Procedural variables used in different methods for labeling dendritic spines have been quantitatively evaluated for their impact on the ability to resolve individual spines in confocal microscopic analyses. In contrast, there have been discussions, though no quantitative analyses, of the potential effects of choosing specific mounting media and immersion oils on dendritic spine resolution.
Here we provide quantitative data measuring the impact of these variables on resolving dendritic spines in 3D confocal analyses. Medium spiny neurons from the rat striatum and nucleus accumbens are used as examples.
Both choice of mounting media and immersion oil affected the visualization of dendritic spines, with choosing the appropriate immersion oil as being more imperative. These biologic data are supported by quantitative measures of the 3D diffraction pattern (i.e. point spread function) of a point source of light under the same mounting medium and immersion oil combinations.
Although not a new method, this manuscript provides quantitative data demonstrating that different mounting media and immersion oils can impact the ability to resolve dendritic spines. These findings highlight the importance of reporting which mounting medium and immersion oil are used in preparations for confocal analyses, especially when comparing published results from different laboratories.
Collectively, these data suggest that choosing the appropriate immersion oil and mounting media is critical for obtaining the best resolution, and consequently more accurate measures of dendritic spine densities.
结构可塑性，如树突棘形态和密度的变化，反映了突触连接性和神经回路的变化。在共聚焦显微镜分析中，已对不同树突棘标记方法中使用的程序变量对分辨单个树突棘能力的影响进行了定量评估。相比之下，对于选择特定封片剂和浸没油对树突棘分辨率的潜在影响已有讨论，但尚无定量分析。
在此，我们提供了测量这些变量对三维共聚焦分析中分辨树突棘影响的定量数据。以大鼠纹状体和伏隔核中的中等棘状神经元为例。
封片剂和浸没油的选择均会影响树突棘的可视化，其中选择合适的浸没油更为重要。这些生物学数据得到了在相同封片剂和浸没油组合下点光源三维衍射图案（即点扩散函数）定量测量的支持。
尽管这不是一种新方法，但本手稿提供了定量数据，证明不同的封片剂和浸没油会影响分辨树突棘的能力。这些发现突出了报告共聚焦分析制剂中使用的封片剂和浸没油的重要性，尤其是在比较不同实验室发表的结果时。
总体而言，这些数据表明选择合适的浸没油和封片剂对于获得最佳分辨率至关重要，从而能更准确地测量树突棘密度。
