Sample Preparation

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===Steps for Sample Preparation===
 
===Steps for Sample Preparation===
  +
====Hydrogel solution====
 
Begin by preparing the [[Solutions#Hydrogel Solution|hydrogel solution]]. This solution can be made in batches and stored in the freezer until needed.
 
Begin by preparing the [[Solutions#Hydrogel Solution|hydrogel solution]]. This solution can be made in batches and stored in the freezer until needed.

Revision as of 00:26, 4 April 2014

Clearing tissue samples using CLARITY begins with introducing hydrogel monomers into the tissue. The monomers are introduced via transcardial perfusion and/or sample incubation with a hydrogel monomer solution.

CLARITY-applicable Tissue Samples

CLARITY can be performed on both freshly-extracted tissue samples as well as fixed tissue samples. The main difference in processing these samples is the incubation time in the hydrogel monomer solution.

While CLARITY was developed for brain tissue, it is applicable to a variety of other tissues as well. Challenges may arise for tissues that are particularly fibrous as they may undergo increased crosslinking and therefore limit the hydrogel porosity during clearing. CLARITY uses fat-collecting micelles to remove light-scattering lipids from the tissue to achieve transparency. If there are other non-fatty light-scattering elements in the tissue, additional clearing using a separate solution to remove those elements may be needed.

Steps for Sample Preparation

Hydrogel solution

Begin by preparing the hydrogel solution. This solution can be made in batches and stored in the freezer until needed.