Electrokinetic Delivery and Extraction of Macromolecules Across Intact Biological Tissues

Technology #16669

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Device used for the electrophoretic movement of molecules throughout intact tissue
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Inventors
Professor Kwanghun Chung
Department of Brain and Cognitive Sciences, MIT
External Link (chunglab.org)
Sung-Yon Kim
Department of Brain and Cognitive Sciences, MIT
External Link (chunglab.org)
Kimberly Ohn
Department of Chemical Engineering, MIT
Evan Murray
Department of Brain and Cognitive Sciences, MIT
Jae Cho
Department of Chemical Engineering, MIT
Managed By
Jon Gilbert
MIT Technology Licensing Officer
Patent Protection

Active transport of charged molecules into, within, and/or from charged matrices

PCT Patent Application WO 2015-154000

Active transport of charged molecules into, within, and/or from charged matrices

US Patent Pending US 2015-0285765

Applications

This invention can be used for the histological examination of intact tissue in both the research and medical fields, enabling rapid evaluation of healthy and disease states. Specific applications include the identification of neuronal network connections, cell type identities, and long-range projections across organs.

Problem Addressed

Labeling and imaging of fine structures within complex, intact systems requires sufficient tissue preservation and light penetration. Permeabilizing tissue via lipid removal often results in the dissolving and/or damaging of fine cellular architecture. The use of a fixative prior to controlled electrokinetic delivery of charged macromolecules allows for the detection and binding of intact, fine structures with multi-round capability. 

Technology

This technology is an electrokinetic device that promotes the rapid flow of charged macromolecules (e.g. detergents, fluorescent probes, antibodies, proteins, nucleic acids) into and out of intact, large-scale biological tissue. A two-step process is used to 1) fix and permeabilize tissue and 2) deliver charged molecules to said tissue. A three-channel design is used whereby the cathode and anode, located in the first and third channels, respectively, lie directly outside two nanoporous membranes that surround the specimen in the second, center channel. These membranes allow for the passage of electric fields and small molecules but restrict macromolecules (e.g. probes of interest) in a size-dependent manner, thus preserving costly reagents and maintaining a steady pH. Multiple probes can be employed at once and separated via pH-based migration. Rotation of the specimen and distribution of electrical force serve to minimize tissue damage. As such, multiple rounds of delivery and extraction are possible in the same tissue sample with minimal damage or protein loss.

Advantages

  • Rapid delivery/extraction of macromolecules: complete lipid removal from intact mouse brain within several hours
  • Capable of multiple functions including lipid removal, fluorescent probe/antigen delivery, and nucleic acid delivery
  • Can perform multiple rounds of delivery/extraction in an individual tissue sample with minimal damage or protein loss
  • Three-channel electrokinetic device conserves expensive reagents while controlling pH and temperature