1/6/2018
Cancer Research & Genetics UK have donated £10,000 to purchase Imaging equipment in the X- Clarity Tissue Clearing System at Cardiff University. The research is led by Professor Matt Smalley at the European Cancer Stem Cell Research based at the University.
X-CLARITY Tissue Clearing system. Revealing tumour composition, architecture and underlying mechanisms in 3D
Tumours are heterogeneous complex environments composed of multiple cell types and matrix proteins. How cells interact and organize within a tumour often hint at the underlying biology that governs tumour development and malignancy. Equally, by monitoring expression levels, activity or localisation of specific proteins within cells also informs on mechanisms and may identify new therapeutic targets. Traditionally, we have relied on immunofluorescence methods to label cells within slices of fixed tissue and then retrospectively build 3D maps of the tissue using specialised confocal microscopy and image analysis tools. This traditional approach requires specialised training and expertise, and is also time consuming.
Recent advances in tissue imaging have led to the development of the CLARITY method (Clear Lipid-exchanged Acrylamide-hybridised Rigid Imaging/Immunostaining/In situ-hybridisation-compatible Tissue-hYdrogel). This method clears or removes masking lipids from whole tissues to make them transparent, exposing the remaining protein and DNA in cells. Because the tissue is fixed in a hydrogel scaffold, this method allows us to examine proteins and therefore cellular structures within preserved 3D tissue architecture. We propose to purchase the X-CLARITY tissue clearing system from Labtech. This equipment provides an all-in-one automated, rapid and efficient tissue clearing protocol, without exposing the user to toxic chemicals or extended incubation steps. Therefore, the protocol is standardised, which saves time and increases robustness. The stained tissue is then imaged in a specialised microscope called a Light sheet (Selective Plane Illumination Microscopy; SPIM, in house at the School of Biosciences). Light sheet microscopy is a time efficient way to rapidly image 3D systems. Together, this approach will allow us to build 3D maps of increasing complexity that reflect the subcellular and cellular networks of a tumour. The CLARITY method can be applied to any solid tumour, including pancreas, breast, brain and bone. This will give us a better understanding of the biology underlying tumour development and help us to identify rare metastatic lesions in secondary organs.