HHH-IDIA
Our Objectives
Imaging under BSL3 conditions offers the unique advantage of monitoring infection processes of unmodified fully infectious BSL3 pathogens in complex and physiologically relevant model systems. However, the required biosafety measures entail practical limitations in organizing, maintaining and using BSL3 microscopy infrastructure as well as user training. This currently impedes the progress and throughput of BSL3 imaging projects.
The main goal of this project is to develop and implement technologies, strategies and measures to overcome these limitations. Consequently, our platform (under the name Heidelberg-Hamburg Infectious Diseases Imaging Association, HHH-IDIA) will be able to offer support and access to advanced microscopy instrumentation for a significantly increased number of BSL3 imaging projects. Furthermore, HHH-IDIA will offer imaging techniques that are not available under BSL3 conditions elsewhere in Germany, in particular light sheet and two-photon microscopy for close-to-physiological model systems, and correlative light- and electron cryo-microscopy, to the research community.
Objective 1:
Enhancing the throughput of BSL3 imaging infrastructure
Imaging under BSL3 conditions is a complex endeavor that needs input from microscopy-, biosafety-, infection biology-, and data analysis experts. The best way to enhance throughput for BLS3 imaging in a safe manner - and hence to enable HHH-IDIA to support an increased number of BSL3 imaging projects - is to establish standardized workflows. Thus, our objective is to design and implement standardized workflows and quality control measures for BSL3 imaging and to continuously upgrade them by incorporating new technical developments.
Objective 2:
Development of and access to BSL3 imaging for close-to-physiological model systems
Imaging technologies, such as selective plane illumination microscopy (SPIM) and two-photon excitation microscopy (TPEM) enable to image physiologically relevant model systems such as organoids, organ explants and live animals. Thus, they are of particular importance for understanding the infection process and the immune reaction to human pathogens. The HHH-IDIA project partners are to our knowledge the only sites in Germany that operate these technologies for research on human BLS3 pathogens. Our objective is to adapt and improve these technologies for close-to-physiological imaging under BSL3 conditions.
Objective 3:
Development of and access to BSL3 cryo-microscopy technologies
Vitrification to produce amorphous ice is a powerful method to preserve biological samples for microscopic imaging. However, the preservation in a native state also means that vitrified BSL3 pathogens do not get inactivated and are still infectious upon reheating and/or mechanical detachment from the sample. Hence, vitrified BSL3 samples need to be handled with the same precaution as live BSL3 samples. Our objective is to develop new technologies for sample handling to enable BSL3 correlative light and electron microscopy under cryo-conditions (cryoCLEM) for vitrified samples with enhanced throughput and minimized sample loss. Furthermore, we will develop and implement workflows for BSL3 cryoCLEM.
Objective 4:
Support for registering BSL3 imaging projects
Regulations and regulatory procedures for scientific studies with BSL3 organisms can differ between states in Germany and permits for carrying out BSL3 experiments are only valid in the state in which the application was submitted. This represents a considerable practical obstacle for a BSL3 imaging infrastructure that wants to serve and be used by researchers from all over Germany and beyond. Our objective is to create a framework that supports researchers navigating the regulatory procedures that are required before a BSL3 imaging project can start.
