-- Prof. Ian Collinson Lab --This was a Wellcome Trust project for which I recommended a new technology to develop a novel biochemical assay. We developed a real-time luminescent assay based on the NanoBiT technology to monitor protein translocation in mitochondria. This has successfully reduced acquisition time from 2 days to minutes, providing high kinetic detail. I then moved to study the contribution of Δψ and ΔpH on protein import of native substrates using this assay. During my time in the lab I managed and supervised a PhD project for optimising the assay in mammalian cells.- yeast & bacteria culture - applikon bioreactor (30L) - protein expression (yeast & bacteria) - in vitro translation systems - protein purification - AKTA systems - molecular cloning (restriction enzymes, Gibson assembly) - protein & vector design - DNA extraction & purification - plasmid PCR - site-directed mutagenesis - Nanoluc - lentivirus production - lipofectamine transfection - enzyme kinetics (assay & modelling) - Python

-- Prof. Michael Duchen and Prof. Gyorgy Szabadkai Lab --Here I was working jointly with GSK Neuroscience R&D (Richard Rutter) to confirm efficacy of pre-clinical drug candidates in inhibiting the mitochondrial permeability transition. I was responsible for development & optimisation of phenotypic assays, where I recommended two novel approaches to increase robustness. Partnering with big pharma helped me to develop time management & prioritising skills in order to regular produce and present data reports, discuss strategies and project directions.- mammalian cell culture - primary cell culture (cardiomyocytes & hepatocytes) - high-content screening - ImageXpress - MetaXpress - high-throughput cellular screening (FLIPR) - confocal microscopy - lipofection - flash-type luciferase - oxygraph - high-resolution respirometry - ImageJ - mitochondrial isolation - plate reader

-- Prof. Andrew Halestrap Lab --In this BHF project I was responsible for characterizing mitochondrial morphology and mitochondrial contact sites by EM upon ischemic injury and correlating it with ROS production & mPTP opening. I jointly lead the collaboration with the EM facility in order to solve practical issues and discuss results. I also taught myself how to use EM imaging process algorithms to accelerate our pipeline and meet deadlines.- animal handling - heart perfusion - perfusion protocols - mitochondria isolation - oxygraph - mitochondrial volume analysis - enzymatic activities - western blot - electron microscopy - light scattering - image processing

-- Paulo Oliveira, PhD Lab --This project consisted of a multi-organ study of doxorubicin cardiotoxicity in a rat model, with special focus on mitochondrial dysfunction. Working with a technician I was responsible for daily handling and drug administration in animals under a sub-chronic protocol. This required me to develop planning and prioritising skills. I used a number of biochemical assay and applied concepts from control flux analysis to study the differential response of doxorubicin.-animal handling - animal experimentation - mitochondrial physiology - western blot - qPCR - mammalian cell culture - enzymatic activity on small high-throughput scale - mRNA extraction (tissues) - in vitro toxicity assays - tissue culture - cell assays - growth assays

Scientific training for fulfilment of degree in biochemistry. Project consisted in studying the mechanism of action of the natural alkaloid berberine in vitro using two distinct toxicological models: isolated mitochondria (hepatic) and a cell line.- mitochondria isolation - mitochondria physiology - HPLC - mammalian cell culture - in vitro toxicity assays - fluorescence microscopy - toxicology profiling