A new advanced method CuRVE, better known as continuous redispersion of volumetric equilibrium is designed to label proteins in intact tissues and organs. The technique is announced by the cooperation of several MIT laboratories related to brain sciences and chemical engineering, resolving a long-waited issue in biological research. The approach to ensure equal distribution of antibody throughout whole organs, has been demonstrated in several tissue types and could significantly enhance the accuracy of cellular studies. The original research paper is published in Nature Biotechnology (doi: 10.1038/s41587-024-02533-4)
Scientists are processing the integration of this approach to upgrade the study level for complex biological units without disturbing natural architecture of tissues and cellular walls.
The research team, led by Kwanghun Chung, associate professor of chemical engineering and neuroscience at MIT, tested the method on various biological samples, including rodent and human tissues.
By controlling the rate of antibody-antigen interactions and enhancing molecular diffusion through stochastic electrotransport, the process enabled uniform protein labeling in a fraction of the time required by conventional methods.
Traditional approaches like immunohistochemistry often fail to achieve uniform protein distribution due to the size of labeling molecules. Antibodies tend to concentrate near the surface while failing to penetrate deeper layers, leading to uneven labeling.
In a statement to MIT News, Mr. Chung compared the challenge to marinating a thick cut of meat, where only the outer layers effectively absorb the marinade. He explained that the large size of labeling molecules makes uniform penetration extremely difficult, requiring prolonged processing times.
The researchers demonstrated that their CuRVE technique, combined with rapid electrophoretic-based labeling using affinity scanning in hydrogel (eFLASH), significantly improved antibody penetration. Tests on an adult mouse brain showed that conventional methods resulted in uneven labeling, while Eflash produced uniform staining across all neurons. Similar success was observed in tissues from other species, including human and tit brain samples, as well as mouse embryos, lungs, and hearts.
Scientists anticipate that CURVE will contribute to the creation of a comprehensive repository of protein expression patterns in different tissue types. This could serve as a reference for studying diseased tissues and refining diagnostic methods. The researchers believe that by enhancing protein visualization at the cellular level, the technique can improve the understanding of various biological processes, potentially influencing future medical and scientific applications.