1. Matrix gel plate operation specification.

Key points for pipette operation:

When spreading the gel, keep the pipette vertical to the top of the inner hole of the 96-well plate to avoid the matrix gel from contacting the wall of the hole;

If bubbles appear, immediately touch the surface of the bubble with the pipette tip to puncture it, and then gently shake the plate to spread the matrix gel evenly on the bottom of the well.

Adjustment of liquid surface flatness:

If the bottom of the matrix gel plate is uneven, you can promote uniform distribution by gently shaking the plate horizontally (or using a shaker).

2. Potential causes of abnormal cell distribution in Transwell chambers.

Operational factors:

The Transwell chamber is placed in the well plate at an angle;

Insufficient cell digestion leads to agglomeration and uneven suspension inoculation;

The matrix gel surface is uneven or there are bubbles.

Experimental design factors:

The cell's own invasion ability is weak;

The Matrigel concentration is too high or the thickness of the gel is too thick, which hinders cell migration;

The cell seeding density is insufficient, or the pore size of the Transwell chamber membrane is too small (such as