CLARITY may seem like an intimidating technique requiring a lot of time, effort, and expensive equipment to get it started and working consistently. However, many of the CLARITY steps can be simplified to make the process easier to get started.

The following recommendations are useful for getting started with CLARITY and getting a feel for performing each step and what to expect from the tissue sample in each stage. Once confident that the technique is working as it should, modifications that may increase complexity can be added. Depending on the application, the simplified CLARITY approach may be all that is needed.

Hydrogel Tissue Embedding

• Use the standard hydrogel solution containing bis-acrylamide. Modifying the hydrogel solution may be useful for certain types of samples, but it is best to start with the standard solution when attempting hydrogel embedding for the first couple times. The standard solution should form a tangible bulk hydrogel around the tissue sample after about three hours of high temperature incubation, which will indicate that the hydrogel embedding are working properly.