Methods for next generation three-dimensional histology for human neural tissues
Novel tissue clearing techniques developed in recent years have enabled labelling and visualisation of neural structures in three-dimensions (3D). However, there have been varying degrees of success in applying these techniques to the human brain tissues. Hence, we have developed a range of techniques for tissue clearing of post-mortem brain tissues. These methods were designed to be easily applied in any standard histology or basic science laboratories. Techniques covered in this protocol include standard immunohistochemistry to enable detailed 3D mapping of neural structures. We have also developed several non-immunohistochemical means of labelling, such as cresyl violet for neuronal quantification, lectin for labelling of blood vessels and DiI retrograde labelling for visualisation of neuronal projections and dendritic spines. The overall processing time for fixed tissue to imaging ranges from 6 hours to approximately 6 months depending on the size of the tissue block and the protocol being followed.
Figure 1
Figure 2
Posted 13 Aug, 2018
Methods for next generation three-dimensional histology for human neural tissues
Posted 13 Aug, 2018
Novel tissue clearing techniques developed in recent years have enabled labelling and visualisation of neural structures in three-dimensions (3D). However, there have been varying degrees of success in applying these techniques to the human brain tissues. Hence, we have developed a range of techniques for tissue clearing of post-mortem brain tissues. These methods were designed to be easily applied in any standard histology or basic science laboratories. Techniques covered in this protocol include standard immunohistochemistry to enable detailed 3D mapping of neural structures. We have also developed several non-immunohistochemical means of labelling, such as cresyl violet for neuronal quantification, lectin for labelling of blood vessels and DiI retrograde labelling for visualisation of neuronal projections and dendritic spines. The overall processing time for fixed tissue to imaging ranges from 6 hours to approximately 6 months depending on the size of the tissue block and the protocol being followed.
Figure 1
Figure 2
© Research Square 2021