Additional file 6: Figure S3. of Light-sheet microscopy for everyone? Experience of building an OpenSPIM to study flatworm development

Abstract

A and B Schematic drawing of laser beam visualized on agarose hanging from above into the water filled acquisition chamber, Also seen in A and B are three alignment steps of the laser beam (1-3); C and D Actual misaligned and aligned laser beams visualized on agarose by removing emission filters and cylindrical lenses; E and F SPIM images (maximum projections) acquired with misaligned and aligned laser beams. The initially visible fuzzy beam is indicated by a bright blue horizontal stripe in between orange arrows. This coarse beam is then brought into focus with the detection objective (step1) and therefore appears as a much thinner laser beam indicated by a blue horizontal stripe in an hourglass-like shape. Note that in this example the focal point of the beam is at this point still shifted to the left (vertical grey line) and need further adjustments (step2). B The laser beam is shifted from the top to a central position within the field of view (step3). (TIF 8563 kb