Die DZL Online Academy bietet insbesondere Nachwuchswissenschaftlerinnen und -wissenschaftlern die Möglichkeit, sich über frei zugängliche Video-Tutorials orts- und zeitunabhängig weiterzubilden. Ein weiteres Anliegen der Academy ist es, die Harmonisierung der Methodenkompetenz über die DZL-Standorte hinweg zu unterstützten. Eine Erweiterung des Angebots ist geplant.

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Assessing lung structure by stereology

Summary

The method of choice for obtaining quantitative (morphometric) data on lung structure in microscopy is stereology. Standards for the quantitative assessment of lung structure by stereology have been published as an official research statement of the American Thoracic Society and the European Respiratory Society (Hsia, Hyde, Ochs, Weibel: Am J Respir Crit Care Med 2010;181:394-418). These two webinars illustrate the principles of stereology and their practical application to the lung, including animal models of human lung disease, according to the ATS/ERS guidelines.

Part 1


Part 2


Videomorphometric Analysis of Hypoxic Pulmonary Vasoconstriction of Intra-pulmonary Arteries Using Murine Precision Cut Lung Slices

Summary

Hypoxic pulmonary vasoconstriction (HPV) is an important physiological phenomenon by which at alveolar hypoxia lung perfusion is matched to ventilation. The major vascular segment contributing to HPV is the intra-acinar artery. Here, we describe our protocol for the analysis of HPV of murine pulmonary vessels with diameters of 20-100 μm.

Webinar accessible via the JoVE Journal webpages

Titelfolie DZLOnlineAcademy PaddenbergEtAlVideomorphometric


Legionella pneumophila Outer Membrane Vesicles: Isolation and Analysis of Their Pro-inflammatory Potential on Macrophages

Summary

Here, we describe the purification of Legionella pneumophila (L. pneumophila) outer membrane vesicles (OMVs) from liquid cultures. These purified vesicles are then used for the treatment of macrophages to analyze their pro-inflammatory potential.

Webinar accessible via the JoVE Journal webpages

Titelfolie DZLOnlineAcademy PaddenbergEtAlVideomorphometric


Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-Cut Lung Slices

Summary

Respiratory diseases in their broad diversity need appropriate model systems to understand the underlying mechanisms and enable development of new therapeutics. Additionally, registration of new substances requires appropriate risk assessment with adequate testing systems to avoid the risk of individuals being harmed, for example, in the working environment. Such risk assessments are usually conducted in animal studies. In view of the 3Rs principle and public skepticism against animal experiments, human alternative methods, such as precision-cut lung slices (PCLS), have been evolving. The present paper describes the ex vivo technique of human PCLS to study the immunomodulatory potential of low-molecular-weight substances, such as ammonium hexachloroplatinate (HClPt). Measured endpoints include viability and local respiratory inflammation, marked by altered secretion of cytokines and chemokines. Pro-inflammatory cytokines, tumor necrosis factor alpha (TNF-α), and interleukin 1 alpha (IL-1α) were significantly increased in human PCLS after exposure to a sub-toxic concentration of HClPt. Even though the technique of PCLS has been substantially optimized over the past decades, its applicability for the testing of immunomodulation is still in development. Therefore, the results presented here are preliminary, even though they show the potential of human PCLS as a valuable tool in respiratory research.

Webinar accessible via the JoVE Journal webpages

Titelfolie Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-Cut Lung Slices