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Fig. 8 | BMC Biology

Fig. 8

From: CLARITY increases sensitivity and specificity of fluorescence immunostaining in long-term archived human brain tissue

Fig. 8

Super-resolution microscopy with STED and dSTORM imaging of synaptic and neuronal markers on human brain tissue after CLARITY. a Synaptic GLUA2 in the cerebral cortex acquired in confocal (upper left) and STED mode (lower right). Scale bar, 1 µm. b Insets (boxed area from (a)) provided in higher magnification unveil postsynaptic membrane morphologies like disk shape as seen in a side view on a synapse (asterisks, left column) and the detailed localization of GLUA2 within a single synaptic spot as seen in a top view on a synapse (right column). A line intensity profile (right part of (b)) across the spot surface demonstrates the intensity fluctuation along the spot (white line in the boxed areas of (b)). Mean intensities (gray values) were normalized on the highest value. a.u. = arbitrary unit. Scale bar, 1 µm. c MAP2 staining in a cerebral cortex section. STED and confocal modes of the same neuron are shown in comparison as in panel (a). Scale bar, 1 µm. d Insets (boxed area from (c)) provided in higher magnification show structural details in the STED mode. The intensity profile across the white lines in the images is plotted on the right. Scale bar, 0.5 µm. Sections for MAP2 and GLUA2 staining were derived from the superior frontal gyrus of perfusion-fixed cases. In (a)-(d), pixel size for confocal images is 50 nm. e Schematic drawing showing expected precision imaging of synaptic compartments in super-resolution images (upper left). SYP/GLUA1 co-staining in STED microscopy. The channels are shown individually (left and middle) and as overlay (right). Here, sections were incubated in hydrogel solution for two days, total clearing time was three days. Scale bars, 200 nm. f SYP/GLUA1 co-staining in dSTORM microscopy (left). An intensity profile across a synapse (rectangle in the left image, width of ten pixels) is shown on the right. Sections were incubated in hydrogel solution for one day, total clearing time was two days. Scale bar, 200 nm. All sections in (e)-(f) were derived from perfusion-fixed cases

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