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

Abstract

Source: Chen, B. et al., Facilitating Cerebral Organoid Culture via Lateral Soft Light Illumination. J. Vis. Exp. (2022).

This video showcases the utilization of a lateral soft light illumination technique to improve the naked-eye visualization of embryoid bodies, thereby facilitating their manipulation and transfer during cerebral organoid culture. It elucidates the setup of soft light illumination, methods for media changes and embryoid body transfer, as well as the procedure for inducing neural differentiation in embryoid bodies for cerebral organoid development.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.

1. Preparation of the soft light lamp (day 1)

1. Use a transparent acrylic board with a thickness of 0.3-0.5 cm in the size of A5 paper. Paste white pads on the front and back of the acrylic plate.
2. Install a row of LED white lights on the edge of the plate so that the lights can enter from the side of the acrylic plate and then shoot out in parallel (Figure 1A-F, Figure 2B).
   NOTE: As the diameters of EBs in the early stage are approximately 200 µm ....

Results

Figure 1: Preparation of the soft light lamp. (A) The power supply and the LED lights were installed on one side of the acrylic board (as shown in the red dotted box). The LED soft light lamp's scattering wavelength is 450-470 nm, luminous flux is.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.2 μm Filter	NEST Biotechnology, China	331001
1000 μL wide-bore pipette tip	Thermo Fisher Scientific, USA	9405163
6-well low adhesion plate	NEST Biotechnology, China	703011	It is used for EBs suspension cultures
Centrifuge	Eppendorf, Germany	5810 R	It can be used for centrifugation of various types of centrifuge tubes, reagent bottles and working plates.
DMEM-F12	Thermo Fisher Scientific, USA	11330032
Heparin	Merck, Germany	H3149
Glutamax supplement	Thermo Fisher Scientific, USA	35050061
Glutamax supplement	Thermo Fisher Scientific, USA	17504044
MEM-NEAA	Thermo Fisher Scientific, USA	11140050
N2 supplement	Thermo Fisher Scientific, USA	17502048
Soft light lamp	NUT	NUT	A simple self made device, refer to supplementary figure 2 for preparation.