Figure 7.

Light-sheet imaging parameters for cellular imaging across the entire mouse brain

(A) Scan configuration for the fastest acquisition. Typically, a 3 × 3 tile scan, using 10% overlap, covers a large enough field of view to encompass the mouse brain. Empirically the left/right illumination, z, channel, x, y acquisition order was found to be the fastest.

(B) The tradeoff between focus depth and waist curvature of the light-sheet. LaVision’s approach to adjusting the sheet width is by effectively adjusting the excitation numerical aperture (NA). A higher NA gives a thinner horizontal focus but with steeper curvature; this gives better center focus, but focus decays rapidly with increasing distance from the center, increasing risk for photobleaching of tissue.

(C) Computational stitching between tiles reduces seam artifacts. The red arrows point to seams in images without (top) and with (bottom) computational stitching.

(D) Sigmoidal blending between stitched left- and right- sided illumination images to produce the final image. Scale bars 250 μm.