To better understand disease mechanisms, we model various lung pathologies using innovative techniques. By exploring their underlying pathophysiology through state-of-the-art phenotyping methods, we aim to pave the way for novel therapeutic strategies.

To precisely investigate the molecular mechanisms driving lung diseases, we assess cell-specific responses to various stimuli (e.g. hypoxia, nitrosative/oxidative stress, cigarette/e-cigarette smoke, inflammatory signals, airborne pollutants, toxins). In addition to traditional two-dimensional (2D) cultures, we employ cutting-edge three-dimensional (3D) culture systems, including air-liquid interface systems, alveolar spheroids, and bronchioalveolar lung organoids. Our expertise enables the generation of clinically relevant disease models and facilitates advanced analyses, such as transcriptomics, proteomics, and functional assays.

Our molecular biology laboratory is equipped with state-of-the-art instruments for investigating the molecular building blocks of cells, including DNA, RNA, and proteins. In addition to performing bulk tissue analysis, we can precisely isolate and analyse specific cell populations and their molecular signatures using advanced tools such as the Laser-assisted Microdissection (LMD) system, Capillary-based Protein Immunoassay (JESS) and the Bio-Plex 200 system. This sophisticated infrastructure enables comprehensive analyses of gene and protein expression, offering insights into the molecular mechanisms driving cellular responses under various physiological and pathological conditions.

To visualize and phenotype our models, we use state-of-the-art imaging systems to obtain high-quality images. The cutting-edge EVOS M7000 Cell Imaging System, an automated platform equipped with high-sensitivity cameras for fluorescence, bright field, and phase contrast imaging, supports exceptional versatility for a variety of applications, including live-cell imaging. This capability provides unparalleled insights into active biological processes in real-time. The Leica STELLARIS 5 confocal microscope features next-generation White Light Lasers (WLL), enabling the simultaneous use of up to eight single excitation lines across the full spectrum. This design offers exceptional flexibility and precision. Its advanced Power HyD detector family delivers superior photon detection efficiency (PDE) and exceptionally low dark noise, ensuring brighter, clearer images with minimal interference. Additionally, the unique TauSense technology integrates application-focused imaging tools based on fluorescence lifetime, allowing researchers to explore the functional roles of molecules within their cellular context. Furthermore, with fluorescence microscopy, we can monitor real-time changes in calcium levels, mitochondrial membrane potential, and cellular membrane potential.

To study the electrical properties of ion channels in cell membranes, our lab is equipped with both a traditional and a fully automated Patch Clamp system. The automated system, in particular, supports high-throughput and high-resolution recordings. Both systems allow measurements of membrane potential and ionic currents, even at the single-channel level. With its unrivaled signal-to-noise ratio, the Patch Clamp system is regarded as the gold standard for measurement of cellular electrophysiological responses. Additionally, the Two Electrode Voltage Clamp (TEVC) technology, combined with a specialized overexpression system, provides precise control of the membrane potential while measuring ionic currents. This capability facilitates detailed analysis of ion channel function and kinetics. Due to its robust signal detection, TEVC is ideal for evaluating the effects of genetic modifications, pharmacological agents, or hypoxia on ion channel activity. Our lab employs a fully automated TEVC system for efficient, reproducible and high throughput ion channel screening.

To measure unpaired electrons, such as reactive oxygen species (ROS), our lab is equipped with an EMXmicro Electron Spin Resonance (ESR) spectrometer. By applying a magnetic field and supplying electromagnetic energy in the microwave frequency range, transitions between electron spin states can be induced. This technique provides critical insights into the molecular and electronic properties of samples. Additionally, we analyze the Raman spectrum of living cells by using RAMAN spectroscopy. This methology employs the inelastic scattering of light to investigate molecular vibrations, rotations, and other low-frequency modes in living samples. When light - typically originating from a laser - interacts with the cellular molecules, a small portion of it is scattered at different wavelengths. This scattering creates a unique spectral fingerprint that reveals the chemical composition and molecular structure of the sample. The highly specific, fingerprint-like Raman spectrum allows to study molecular interactions and evaluate physiological or metabolic responses to environmental stressors, including mitochondrial complex responses to acute hypoxia.

Mitochondria are multifunctional organelles involved in a variety of critical cellular processes, including metabolic pathways, oxygen sensing, redox balance, stem cell differentiation, senescence, aging, cell death, and immunity. They are the primary producers of cellular energy (ATP), achieved by coupling electron transport through the electron transport system (ETS) and oxygen consumption (respiration) to oxidative phosphorylation (OXPHOS).  We have established a core facility specializing in the characterization of mitochondrial function in the lungs and heart. Our lab is equipped with three state-of-the-art Oxygraphs (Oroboros-O2k), enabling precise measurement of oxygen consumption (respiration), redox state of coenzyme Q, NAD(P)H, hydrogen peroxide production, mitochondrial membrane potential, and calcium levels. Additionally, we utilize two Seahorse devices to facilitate high-throughput analysis of mitochondrial respiration. To comprehensively study mitochondrial function under physiological and pathological conditions, we employ blue native polyacrylamide gel electrophoresis (BN-PAGE) for mitochondrial complexome analysis. We also quantify mitochondrial DNA relative to nuclear DNA, measure mitochondrial transcripts and mitophagy markers, and examine mitochondrial networks using confocal microscopy.