Description
It has long been a major challenge to acquire high-resolution image data from relatively large and complex biological samples (e.g., intact embryos, whole organs of animal models and thick tissue sections) to obtain a systems view of the biological question being studied. Most biological tissues are non-transparent; this is mainly attributed to the lipid bilayer that makes up the cellular membrane in each of the rather packed biological cells within the tissue. Light scattering at the lipid-water interphase (due to differences in refractive index) causes serious problems for imaging, making deep-tissue imaging extremely challenging (the penetration depths in such tissue using confocal and multi-photo imaging are only ~100 and ~500 µm, respectively). In 2013, an electrophoretic tissue clearing (ETC) technique, termed CLARITY, was introduced by Chung and colleagues (Nature 497, 332-337 (2013)) at Stanford University. Since then, they and numerous other research groups have shown how CLARITY could address most of these challenges using a wide variety of biological tissues.
In brief, the CLARITY technique involves the following steps:
- Infusion of hydrogel monomers to the tissue
- Polymerisation to construct a tissue-hydrogel hybrid construct
- Electrophoretic tissue clearing (ETC) to remove lipids in the sample, while keeping the biomolecules and fine structures largely intact
Following ETC, the tissue-hydrogel hybrid would become optically transparent and chemically accessible to deep-tissue antibody labelling and imaging. For example, an intact mouse brain (after ETC) could be immunolabelled and subsequently imaged with light sheet fluorescence microscopy (LSFM), multiphoton microscopy (MPM) or even single-photon confocal microscopy.
Hourly rate
- Internal users: $26
- Collaborators: $52
- External users: $130
Location
Y206
Staff-in-charge
Dr. Michael Yuen
Senior Scientific Officer
- Y208
- 34008788
- michael.yf.yuen@polyu.edu.hk
Dr. Alan Leung
Scientific Officer
- Z215
- 34008984
- alan.sl.leung@polyu.edu.hk
