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Herramienta para producir tejidos transparentes. Una nueva visión histológica en 3D


Oferta Tecnológica
Una pyme francesa especializada en histología, citología y biología molecular ha desarrollado un método para producir estructuras resistentes y tejidos transparentes y por lo tanto susceptibles de investigación microscópica detallada de sus partes funcionales constituyentes. Este método ofrece un nuevo acceso y funcionalidad para comprender mejor la estructura de los tejidos y la relación entre estructura y función. Los investigadores pueden estudiar la red celular 3D implicada en enfermedades y trastornos, centrándose en los tejidos dañados y sin perder la perspectiva global. La empresa busca socios europeos con el fin de establecer acuerdos de cooperación técnica, servicio e investigación.


Clearing tool, a new histological view in 3D
French SME is specialized in histology and molecular biology. Clearing method enables to produce transparent whole tissues providing new accessibility and functionality to get a better understanding of tissue structure and structure-function relationship.
Researchers can study the 3D cellular network implicated in diseases and disorders, focusing on damaged structures without losing global perspective.
They are looking for european partners for technical, services and research agreement.
A French company specialized in histology, cytology and molecular biology has over 10 years´ experience (SME with 35 employees). Its mission is to help and facilitate their customers to develop their pharmaceutical products, medical devices, diagnosis tools, toxicology test, control compliance on agri-food products...

Tissue clearing was developed by Karl Deisseroth and coll. at Stanford University School of Medicine (US). On November 29 2015, he was awarded the breakthrough Prize in Life Sciences.

Many current techniques for tissue sectioning, imaging and analysis are time-consuming, and they present major limitations for 3D tissue reconstruction.

Clearing method produces resistant structures and transparent whole tissues and thus amenable to detailed microscopic investigation of its constituent functional parts. Series of chemical treatments must be applied to achieve transparency. Biomolecules in intact tissues are covalently linked to hydrogel network preserving structural integrity and molecular information. Once the hydrogel network in place, the unattached lipid membranes can be removed without causing structural damage or loss to the intact embedded tissue. The sample is bathing in ionic detergents and subjected to electrophoresis. Enhanced electrodes generate a uniform electric field which breaks lipid bilayers, leaving behind a stable and transparent tissue-hydrogel hybrid that is chemically accessible for molecular phenotyping. The original proteins and nucleic acids are left in place. Protein structure has to be placed in a transparent scaffolding which preserves it. Buffer circulation ensures temperature control with an active cooling system and elimination of electrophoretic waste. After this step, tissues can be restored to its initial dimension with a final step of incubation in refractive index matching solution.

See attached an example of clearing result on brain tissue.

This technology can be coupled with immunostaining. Indeed specific antibodies labeled with fluorescent dyes can highlight anatomic area. Specific molecular field can be detected which could improve optical and molecular access. To analyse the labeled clarified samples, microscopy methods like confocal microscopy, Light Sheet microscopy and COLM are required.

This technology can be convenient for all institutes involved in neurological researches like Alzheimer´s disease or those using mouse spinal models, multiple sclerosis models. One of the strengths of this technology is to follow the movement of a molecule in organism.

See attached an example of using the clearing technology. Blood vessels in a mouse kidney are stained in green with specific antibodies anti-SMA (Smooth Muscle Actin). Extracellular matrix is stained in red with specific antibodies anti-collagen IV. This technology allows identifying potential interactions, mapping the biological structures in 3D.

Despite, the technology is still relatively young, many efforts have been made to optimize clarifying protocols. Immunostaining must be developed internally, it could be considered to develop specific labeling in collaboration with a partner.

The company is looking for european academic research institutes, biotechnology labs, CROs (Contract Research Organisation) and startups in health and biotechnology sectors for technical cooperation agreement, services agreement and research cooperation agreement.

For technical cooperation agreement and services agreement, they would like to expand this technology of tissue clearing and adapt protocols to other fields of research than neuroscience, when a 3D approach is relevant for such pathology studies.

For research cooperation agreement, the company has several successfully experiences in collaborative projects. They are interested in participate in european projects as member of a consortium for example.
Advantages and Innovations:
Tissue clearing makes resistant structures and transparent whole tissues ready for multiple rounds of antibody labeling and imaging. Subsequently imaging of transparent organs is a powerful method to analyze fluorescent labeled cells and molecules in 3D, in healthy or diseased organs. This technology can be useful for many researches going from tracing neural network to exploring the relationship between structure and function with a global perspective. This enables 3D vision of large tissues at single-cell resolution.

Contrary to manual clearing methods, they propose an automated which can ensure gain of time, efficiency and reproducibility.

This technology can also be proposed on various tissues and not only brain or animal tissues.
Stage of Development:
Already on the market

Partner sought

Type and Role of Partner Sought:
- Type of partner sought: european academic research institute, biotechnology lab, CRO, start up.
- Specific area of activity of the partner : Health, biotechnology
- Task to be performed by the partner sought: In vivo study, Innovative therapy development


Type and Size of Client:
Industry SME 11-49
Already Engaged in Trans-National Cooperation:
Languages Spoken:


Technology Keywords:
06002002 Biología celular y molecular
06001005 Diagnósticos, diagnosis
09001002 Métodos e instalaciones de ensayo / análisis