Agarose Cube for Cleared Samples
================================

Procedure
---------

1. If the samples are already dehydrated, must do a rehydration step:

   - 100% Methanol - 10 minutes.
   - 75% Methanol - 5 minutes.
   - 50% Methanol - 5 minutes.
   - 25% Methanol - 5 minutes.
   - PBS - 3x 5 Minutes.

2. Embed the sample in an agarose cube.
3. Dehydrate with methanol gradient.

   - 25% Methanol - 5 minutes.
   - 50% Methanol - 5 minutes.
   - 75% Methanol - 5 minutes.
   - 100% Methanol - 10 minutes.
   - 50/50 BABB/Methanol Overnight, mutating at 4 degrees.

4. BABB.

Notes and Cautions
------------------

- If you go directly from 100% methanol to BABB, the agarose cube will collapse.
- Numbers below are for smaller samples.
- Duration may need optimization on larger specimens.
- Percentage?
- Low-melting temperature?

Related Protocols
-----------------

- :doc:`babb-2-0`