A Technique to Optimize Cerebral Organoid Culture Using Lateral Soft Light Illumination

Take a lateral soft light illumination setup containing an acrylic board fitted with white pads and white LED lights installed on one edge.

Position a low-adhesion multi-well plate containing embryoid body formation medium in front. 

Add the embryoid bodies, or EBs to the plate.

The LED light enters from the side of the acrylic board and emits parallel beams, laterally illuminating the EBs and enabling naked-eye visualization.

Rotate the plate to induce a swirl flow, converging the EBs to the center.

Replace the spent medium with fresh EB formation medium.

Collect the EBs using a wide-bore pipette tip.

Hold the pipette upright to allow the EBs to settle by gravity.

Transfer the EBs to another low-adhesion plate containing a neural induction medium.

Touch the tip to the medium. The EBs sink due to liquid surface tension.

Specific factors in the medium initiate neural differentiation in the EBs, forming a neuroepithelial layer.