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Automated Transmission-Mode Scanning Electron Microscopy (tSEM) for Large Volume Analysis at Nanoscale Resolution

Figure 3

Quality comparison of images acquired on tSEM vs. TEM.

Serial section images from the middle molecular layer of the hippocampal dentate gyrus acquired on a TEM (A1–3) and tSEM (B1–3). They were taken as 8-bit grayscale images, and the brightness and contrast were then adjusted to match the images from the two different EM platforms. A1–3: An obliquely cut dendrite (den1) gives rise to a mushroom-shaped spine (sp1) with postsynaptic density (PSD), making a synapse with an axonal bouton (b) containing synaptic vesicles (SV). This bouton also makes a synapse with another spine (sp2). These spines and bouton are wrapped around by an astrocytic process that contains glycogen granules (G) and polyribosomes (PR). A tubule of smooth endoplasmic reticulum (SER) and mitochondrion (mito) are located in the dendritic shaft. Cross-sectioned microtubules (mt) are clearly visible in an adjacent dendrite (den2), which also contains a mitochondrion (mito). B1–3: A mushroom-shaped spine (sp) on a dendrite (den) makes a synapse with a thickened PSD on an axonal bouton (b) containing synaptic vesicles (SV). A tubule of SER is visible at the base of this spine, along with a cluster of polyribosomes (PR). Cross-sectioned microtubules (mt) are also visible in this dendrite, which also contains a mitochondrion (mito). Clusters of polyribosomes (PR) are visible adjacent to a mitochondrion (in B2–3). Glycogen granules (G) are found in a neighboring astrocytic process (a). The original pixels are retained in all images in this figure. Only brightness and contrast were adjusted to match images acquired on the two EM platforms.

Figure 3

doi: https://doi.org/10.1371/journal.pone.0059573.g003