Abstract
The process through which neurons are labeled is a key methodological choice in measuring neuron morphology. However, little is known about how this choice may bias measurements. To quantify this bias we compare the extracted morphology of neurons collected from the same rodent species, experimental condition, gender distribution, age distribution, brain region and putative cell type, but obtained with 19 distinct staining methods. We found strong biases on measured features of morphology. These were largest in features related to the coverage of the dendritic tree (e.g., the total dendritic tree length). Understanding measurement biases is crucial for interpreting morphological data.
Highlights
There are many techniques through which neuron morphologies may be imaged
This suggests that morphologies obtained by different staining methods seldom agree with each other
Morphology varies by brain region, cell layer, species, and age (Scheibel and Jacobs, 2003; Elston, 2007; Spruston, 2008; Elston and Fujita, 2014)
Summary
There are many techniques through which neuron morphologies may be imaged. In Golgi staining, silver nitrate is introduced to fixed tissue, and the metal is taken up by a sub-population of neurons through a mechanism that remains largely uncharacterized. The method can be subdivided into Rapid Golgi, Golgi-Kopsch, and Golgi-Cox (Koyama, 2013) and each version labels a subset of neurons. Other heavy metals such as osmium and lead can instead be used for dense labeling, which is popular for EM data (Watson, 1958; Tapia et al, 2012). Because it is relatively simple to perform compared to other methods, heavy metals are a popular staining method for fixed tissues
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