SHIELD
In this step, we will be preparing the sample for subsequent processing. Initial fixation with paraformaldehyde is required and is followed by an additional epoxy-based fixation step we call SHIELD. You can read more about the technique in this 2018 paper by Young-Gyun Park et al..
The SHIELD fixation is split into 2 steps using a method called SWITCH. The SWITCH method is based on this 2015 paper by Evan Murray et al.. During the initial SHIELD OFF step, the epoxy diffuses into the sample without crosslinking. Once the sample is moved to the SHIELD ON Solution, the epoxy is activated and crosslinking is initiated inside the sample.
[Note] If you have already completed the SHIELD Epoxy Perfusion Protocol, this step is already completed and you can continue to Delipidation.
There are several variants to the protocol, depending on your particular samples. Click the buttons below to move to the desired variant.
Use this protocol if you have thin slices (anything < 500 µm) in thickness. Delipidation steps are included.