Journal: Cells

Article Title: Comparison of Multiscale Imaging Methods for Brain Research

doi: 10.3390/cells9061377

Figure Lengend Snippet: Comparison of different microscopy platforms used to image the same specimen.

Article Snippet: Many newly introduced commercial microscope platforms testify to this development, including Minflux [ ], Super-Resolution Spinning Disc (SoRa) microscopy [ ], single-molecule based Nanoimager ONI [ ], Abberior STED facility line [ ], Abbelight SAFe 180/360 [ ], nanoFLeye [ ], CODIM imager [ ] or Re-scan microscopy platforms [ ].

Techniques: Comparison, Microscopy

Journal: Cells

Article Title: Comparison of Multiscale Imaging Methods for Brain Research

doi: 10.3390/cells9061377

Figure Lengend Snippet: Comparison of various super-resolution microscopy methods in tissue imaging. A 90 × 90 × 50 ( x / y / z ) µm 3 3D region of mouse brain immunofluorescently labeled with antibodies against Homer1 (Homer; red), Shank2/ProSAP1 (Shank2; green) and MAP2 (white) was imaged using an array detector (Airyscan) microscope ( A1 ), a Lattice-SIM super-resolution microscope ( B1 ) and a STED super-resolution microscope ( C1 ), respectively. DNA was counter-stained with DAPI (blue). Please note that A1 , B1 , and C1 show 3D reconstructions of deconvolved image stacks. ( A2 ) shows a single optical section of the A1 image stack after processing and deconvolution. A subregion of this image slice (white box in A2 ) is shown enlarged as a merged view as well as in monochrome individual channels ( A3 – A6 ). ( B1 ) The same tissue sample used in A1 was imaged at an adjacent position employing Lattice-SIM super-resolution microscopy. ( B2 ) shows a single optical section of the image stack shown in B1. A subregion of this section was selected (white box) for display as enlarged views of the merged as well as individual channels ( B3 – B6 ). ( C1 ) The same tissue sample used in A1 and B1 was imaged at an adjacent position employing STED super-resolution microscopy. ( C2 ) shows a single optical section of the image stack shown in C1 after deconvolution. A subregion of this section was selected (white box) for display as enlarged views of the merged as well as individual channels ( C3 – C6 ). Note the higher resolution of the STED microscopy (bar in C6 , 200 nm instead of 500 nm, as in A6 and B6 ).

Article Snippet: Many newly introduced commercial microscope platforms testify to this development, including Minflux [ ], Super-Resolution Spinning Disc (SoRa) microscopy [ ], single-molecule based Nanoimager ONI [ ], Abberior STED facility line [ ], Abbelight SAFe 180/360 [ ], nanoFLeye [ ], CODIM imager [ ] or Re-scan microscopy platforms [ ].

Techniques: Comparison, Super-Resolution Microscopy, Imaging, Labeling, Microscopy, Staining

Journal: Cells

Article Title: Comparison of Multiscale Imaging Methods for Brain Research

doi: 10.3390/cells9061377

Figure Lengend Snippet: Comparison of various microscopy methods in 3D synapse imaging of mouse brain tissue. ( A ) shows single optical sections of the image stacks described in and ((Mouse brain immunofluorescence labeling of Homer1 (Homer; red), Shank2/ProSAP1 (Shank2; green) and MAP2 (white); DNA counterstained with DAPI (blue)). Single focal accumulations of Homer and Shank2/ProSAP1, likely representing PSDs were selected from each section (white arrows) and displayed as orthogonal views in B . SIOS, structured illumination optical sectioning (ApoTome); SPDM, spinning disc confocal microcopy; SIM, structured illumination microscopy; STED, stimulated emission depletion. Please note, that 2D-SIM and 2D-STED was employed here, while the HyVolution and Airyscan stacks were deconvolved resulting in improved z-resolution. Bars, 1 µm ( A ) and 500 nm ( B ).

Article Snippet: Many newly introduced commercial microscope platforms testify to this development, including Minflux [ ], Super-Resolution Spinning Disc (SoRa) microscopy [ ], single-molecule based Nanoimager ONI [ ], Abberior STED facility line [ ], Abbelight SAFe 180/360 [ ], nanoFLeye [ ], CODIM imager [ ] or Re-scan microscopy platforms [ ].

Techniques: Comparison, Microscopy, Imaging, Immunofluorescence, Labeling

Journal: Cells

Article Title: Comparison of Multiscale Imaging Methods for Brain Research

doi: 10.3390/cells9061377

Figure Lengend Snippet: Methods to improve deep tissue imaging and image processing. ( A – C ) Influence of an objective correction ring on image quality. Mouse brain tissue immunofluorescently labeled for Homer (red) and Shank2 (green). DNA was counterstained with DAPI (white). Imaging was performed in an area 40 µm deep within this tissue section employing point-scanning confocal microscopy ( A ). The same optical section was imaged sequentially with increased settings of a motorized correction ring of a 93× glycerol objective. The signal intensity at a selected synapse of Homer fluorescence (boxed in A ) dependent on correction ring setting is shown in B . ( C ) Quantitation of signal intensity vs. motorized correction ring setting (motCORR) as shown in B ( n = 5 per data point). ( D ) Effect of AO: Representative z-STED microscopy images of Homer (magenta, gp-anti-Homer-STAR 635P immunolabeled) in the mouse brain tissue without (left, AO off) and with (right, AO on) AO correction on the bottle-shaped STED laser (added wavefront distortion in the upper right insets, arbitrary color scale from 0 (blue) to maximum (yellow)) with intensity profiles along the lines in-between the arrows. Scale bar, 1 µm. ( E ) Acceleration of deconvolution by CUDA graphics card. Image stacks of various thickness of the mouse brain tissue as shown in A were acquired on a confocal microscope. Deconvolution of the stacks was performed with or without employment of a CUDA graphics card. The time required for deconvolution was measured ( n = 3) and plotted versus the file size of the image stack.

Article Snippet: Many newly introduced commercial microscope platforms testify to this development, including Minflux [ ], Super-Resolution Spinning Disc (SoRa) microscopy [ ], single-molecule based Nanoimager ONI [ ], Abberior STED facility line [ ], Abbelight SAFe 180/360 [ ], nanoFLeye [ ], CODIM imager [ ] or Re-scan microscopy platforms [ ].

Techniques: Imaging, Labeling, Confocal Microscopy, Fluorescence, Quantitation Assay, Microscopy, Immunolabeling