In this webinar, Dr. Colin Chu (UCL Institute of Ophthalmology, UK) presents the integration of the ARIA fluidics device and THUNDER imager developed in the Germain laboratory at the NIH. Additionally, Dr. Chu describes how he extended the automated IBEX method, developed with James Marr, Andrea Radtke, and Ziv Yaniv, to the retina with his team at UCL. Image credits: Stained glass window cover slide and promotional material (Andrea Radtke and Stefan Uderhardt), lymph node IBEX images (Andrea Radtke), jejunum IBEX images (Andrea Radtke and Colin Chu), skin automated IBEX images (Hiroshi Ichise and Andrea Radtke). The original IBEX method and image alignment software was developed by Drs. Andrea Radtke and Ziv Yaniv at the NIH.
Multiplexed antibody-based imaging enables the characterization of cellular interactions, spatial relationships, and histological patterns in tissues. These techniques require considerable resources and domain expertise for implementation, limiting their widespread adoption. To overcome these challenges, we developed Iterative Bleaching Extends multi-pleXity (IBEX), an open-source method that can be completed at relatively low cost in 2-5 days by biologists with basic laboratory skills. In addition to cost-effective techniques such as IBEX, community initiatives are needed to extend the reach of multiplexed tissue imaging. The IBEX Imaging Community is an international group of scientists committed to sharing knowledge related to multiplexed imaging in a transparent and collaborative manner. Organ Mapping Antibody Panels (OMAPs) are community-validated resources that save time and money, increase reproducibility, and support the construction of a Human Reference Atlas. This webinar will provide guidelines for the successful application of IBEX to a wide range of scientific questions using diverse imaging platforms. A significant amount of time will be devoted to the following topics: optimal tissue processing, working with fixed frozen and FFPE specimens, identifying appropriate reagents, and constructing organ atlases. Team science initiatives will be highlighted to encourage attendees to join and accelerate discovery together.
Multiplexed antibody-based imaging empowers the study of complex cellular phenotypes in situ by enabling the evaluation of dozens of protein biomarkers in a single tissue section. Iterative Bleaching Extends multi-pleXity (IBEX), is an antibody-based staining and chemical bleaching technique that allows comprehensive profiling (65+ parameters) in diverse tissues. It is compatible with 250+ commercially available antibodies and 16 unique fluorophores. It can be adopted by different imaging platforms at a low cost, and the experiment can be done in 2-5 days by biologists with basic laboratory skills. In this webinar, we will go over methods to study normal and malignant lymph nodes and topics related to multiplexed tissue imaging, namely: optimal tissue processing, working with fixed frozen and FFPE specimens, overcoming autofluorescence, identifying appropriate reagents, validating antibodies, and creating organ mapping antibody panels (OMAPs).