Tissues are typically characterized by preparing thin tissue sections, staining with histological stains or immunohistochemistry, and visualization via brightfield or fluorescent microscopy. While this has been the gold standard for over a century, this approach has significant shortcomings when it comes to analyzing complex and heterogeneous tissue structures such as tumors, the brain, or blood vessel networks. As a metaphor, imagine trying to diagnose the problem with the engine of a car if you only had a cross section to work with. In order to analyze complex structures using the traditional histology paradigm, costly and time-consuming serial sectioning must be conducted, or else using stereological techniques to calculate inferential aspects about the whole tissue structure.
Instead, imaging thick tissue specimens and whole-mounts in 3D through the use of tissue clearing, fluorescent labeling and confocal or light sheet microscopy allows for tissues to be characterized intact in their entirety. By pairing tissue clearing with molecular probes or immunolabeling, select biomarkers, cells, and regions can be reconstructed in 3D for analysis. To visualize and characterize these complex features, Visikol has a portfolio of patented tissue clearing reagents, tissue processing techniques and the purpose-built 3Screen image analysis software tools. Visikol was the first company founded to utilize tissue clearing and 3D imaging for research. We are experts that have been working in this space since 2012, and have worked on countless projects with hundreds of universities, biotech companies, and pharmaceutical companies.
Send us your tissues for imaging and analysis
We at Visikol are experts in bio-imaging. We provide tissue imaging services for thick tissue sections, whole mounts, as well as for histological sections. We provide end-to-end services for tissue processing, labeling, imaging, and data analysis.
Transforming a 3D tissue into 3D image sets and mining the data for actionable insights requires significant expertise in tissue processing, immunolabeling, 3D microscopy, and data processing. This wide array of domain experiences makes adopting 3D tissue imaging challenging for many researchers who do not have the appropriate skills and resources. Clients send us tissues of any size for imaging and we deliver image data and analyzed data sets and reports back to Clients to help them address their research questions.
Antibody validation for 3D
At Visikol, we have developed a methodological approach to optimizing deep antibody labeling such that we can achieve uniform labeling across thick tissues while preserving histological integrity. We select only antibodies validated for immunohistochemistry, and we validate each antibody for whole mount immunolabeling.
3D imaging approach
While many publications detail whole mouse brain imaging and other large tissue volumes imaged intact, this is often impractical to address specific research questions. Preparing tissues into 1-2 mm sections dramatically reduces labeling time and simplifies the imaging process, significantly reducing costs. This sample workflow enables us to provide an unprecedented level of 3D tissue imaging throughput to our Clients. On request, we can accommodate larger tissues, utilizing immersion objectives, light sheet microscopy and other imaging modalities though if a specific research question requires them.
Tissue clearing approach
At Visikol, we have developed the Visikol HISTO tissue clearing technique for rendering tissues transparent. This technique is easy-to-use, preserves tissue integrity, morphology, and histology, and is compatible with molecular probes and immunolabeling, as well as fluorescent proteins. Most importantly, Visikol HISTO is rapid and requires no specialized equipment.
3D image analysis
While some of our Clients require only raw data files from our imaging services, we also offer 3D image analysis services using our purpose-built 3Screen™ platform to analyze and quantify 3D image sets. Instead of processing these data sets as Z projections, we process the entire tissue volume. This makes it possible to evaluate each cell within a tissue on an individual basis, making to possible to, for example, quantify cell types or biomarker expression, quantify the co-localization of multiple biomarkers, map blood vessel networks, examine the distribution of cells within and outside of solid tumors, and many other quantitative endpoints.