This is a protocol preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Method Article
Hei Ming Lai
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong
Corresponding Author
Alan King Lun Liu
Neuropathology Unit, Imperial College London
Corresponding Author
Wutian Wu
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong
Corresponding Author
Steve Gentleman
Neuropathology Unit, Imperial College London
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
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.
Keywords
Tissue clearing, immunofluorescence, confocal imaging, neuroscience, neuropathology
Figure 1
Figure 2
The authors declare no competing financial interests
Loading...
Version History
CURRENT STATUS: Posted
Version 1
Posted 13 Aug, 2018
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Associated Publications
Next generation histology methods for three-dimensional imaging of fresh and archival human brain tissues
by Hei Ming Lai, Alan King Lun Liu, Harry Ho Man Ng, +7
Nature Communications (18 May, 2018)
Method Article
Hei Ming Lai
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong
Corresponding Author
Alan King Lun Liu
Neuropathology Unit, Imperial College London
Corresponding Author
Wutian Wu
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong
Corresponding Author
Steve Gentleman
Neuropathology Unit, Imperial College London
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 13 Aug, 2018
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
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
Loading...
Associated Publications
Next generation histology methods for three-dimensional imaging of fresh and archival human brain tissues
by Hei Ming Lai, Alan King Lun Liu, Harry Ho Man Ng, +7
Nature Communications (18 May, 2018)