Journal: bioRxiv

Article Title: Light-sheet microscopy with isotropic, sub-micron resolution and solvent-independent large-scale imaging

doi: 10.1101/605493

Figure Lengend Snippet: (a) Working principle of axially swept light-sheet microscopy: a thin light-sheet is scanned in its propagation direction, the rolling shutter readout of an sCMOS camera, adjusted to the size of the beam waist, is tightly synchronized to the light sheet scan. (b) Experimentally measured PSF of conventional light-sheet fluorescence microscopy (LSFM), Airy scan confocal microscopy, Lattice light sheet microscopy (LLSM) and ctASLM. The LLSM was using the same detection objective as the ctASLM system. (c-d) Maximum intensity projections of CLARITY cleared cortical Thy1-eYFP neurons as imaged by ctASLM. Zoomed in view of the rectangular region in (d) when imaged using, (e) LSFM vs. (f) ctASLM. All images show Raw data, no deconvolution was applied, with the exception of the Airy Scan PSF. Scale bars are: (b) 3 microns, (c,d) 100 microns; (f) 20 microns.

Article Snippet: The detection arm consisted of an identical microscope objective (Cleared Tissue Objective, Advanced Scientific Imaging), a tube lens (ITL200-A, ThorLabs), a 4-position filter wheel (LB10-WHS4 and a Lambda 10-3, Sutter Instruments), and a sCMOS camera (Flash 4.0, Hamamatsu Corporation).

Techniques: Microscopy, Fluorescence, Confocal Microscopy

Journal: Nature methods

Article Title: Light-Sheet Microscopy of Cleared Tissues with Isotropic, Subcellular Resolution

doi: 10.1038/s41592-019-0615-4

Figure Lengend Snippet: (a) Working principle of axially swept light-sheet microscopy: a thin light-sheet is scanned in its propagation direction. The rolling shutter readout of an sCMOS camera, adjusted to the size of the beam waist, is tightly synchronized to the light sheet scan. (b) Experimentally measured PSF of Airy scan confocal microscopy, conventional light-sheet fluorescence microscopy (LSFM), and ctASLM. LSFM and ctASLM PSFs are shown for NA 0.4 (indicated with “1”) and 0.7 objectives (indicated with “2”). (c-d) Maximum intensity projections of CLARITY cleared cortical Thy1-eYFP neurons as imaged by LSFM and ctASLM, respectively. (e) An XY view of a section 2.5mm from the top surface of a Thy1-eGFP PEGASOS cleared brain. (f) Zoom-in of the red box with a rendered neuron. Insets provide magnified views of synaptic spines. (g) Volume rendering of a mouse kidney labeled with Flk1-GFP. Inset shows a 3D rendering of endothelial cells in one selected glomerulus. (h) Maximum intensity projection of a mouse tibia bone marrow plug. (i) Zoom-in of magenta box shows alpha-catulin-GFP+ cells (green, Alexa Fluor 488), Lepr+ niche cells (red, tdTomato) and cKit+ cells (grey, Alexa Fluor 647). Alpha-catulin-GFP+/cKit+ hematopoietic stem cells (HSC) are labeled with cyan spheres. (j&k) are the volume-rendering of boxed region in (h), showing progenitor cells in grey and niche cells in red. All biological data acquired with NA 0.4 objectives. Scale bars are: (b) 3 μm, (c,d) 20 μm, (e) 1 mm, (g) 500 μm, (h) 1 mm, (i) 200 μm, (j) 40×40×40 μm 3 , (k) 5 μm.

Article Snippet: The detection arm consisted of an identical microscope objective (Cleared Tissue Objective, Advanced Scientific Imaging), a tube lens (ITL200-A, ThorLabs), a 4-position filter wheel (LB10-WHS4 and a Lambda 10–3, Sutter Instruments), and a sCMOS camera (Flash 4.0, Hamamatsu Corporation).

Techniques: Microscopy, Confocal Microscopy, Fluorescence, Labeling