HiTS Seminar: Sinem Saka, PhD, 10:00 am, Thursday, September 19, 2019
Wyss Institute for Bioinspired Engineering
Multiplexed and amplified protein imaging in tissues by Immuno-SABER
Mapping the molecular composition of individual cells in their native environment is critical to understand the cellular and subcellular organization of tissues in healthy and diseased states. However, conventional fluorescence imaging methods offer limited (typically <5) multiplexing due to spectral overlap. Sensitivity is the other limiting factor owing to high autofluorescence and scattering in the tissue samples, making it difficult to visualize low abundance targets. To achieve the desired high sensitivity and throughput, a clear need exists for a robust and scalable signal amplification method compatible with rapid multiplexing. To address this challenge, we developed a new in situ protein imaging method, Immunostaining with Signal Amplification By Exchange Reaction (Immuno-SABER), which achieves highly multiplexed signal amplification via DNA-barcoded antibodies and orthogonal DNA concatemers generated by primer exchange reactions (PER). SABER offers independently programmable signal amplification without in situ enzymatic reactions, and intrinsic scalability to rapidly amplify and visualize a large number of targets when combined with fast exchange cycles of short fluorescent strands. We demonstrated 5–180-fold signal amplification in diverse cell and tissue samples and simultaneous application for 10 different proteins using standard equipment and workflows. The SABER approach can also be combined with expansion microscopy to enable rapid, multiplexed super-resolution tissue imaging or can be adapted for fluorescence in situ hybridization for multiplexed imaging of RNA and DNA targets (SABER-FISH). SABER presents an effective and accessible platform for multiplexed tissue imaging with high sensitivity and throughput, and can be enabling for a broad range of applications including molecular atlases, biomarker discovery, and digital pathology.
10:00 am – 11:00 am
Thursday, September 19, 2019
Harvard Medical School
200 Longwood Avenue
Warren Alpert Bldg, Room 563
Boston, MA 02115
Hosted by Kenichi Shimada, Ph.D.