Quantitative cell biology - 4PMBBPH1

Goal(s)

Using illustrating examples, lectures will explain the dynamical organization of living cells, explore different techniques used to study cell functions and show the complementarity between experimental approaches and different scales.

Note that the written course is not taught in the classes. It should nevertheless be read and learned. During tutorials, the course will be used to understand articles and develop critical thinking and creative thinking in the biomedical field.

In these practicals the student learns eucaryote cell culture techniques. Qualitative and quantitative analysis methods to evaluate cellular function are taught.

Content(s)

• Lectures
  • Membrane and cell compartments
  • Gene expression in eukaryotes
  • Cytoskeleton dynamics
  • Membrane traffic
  • DNA replication, repair and recombination

• BE : Scientific article analysis
  • Molecular techniques for cell study: fluorescent protein expression, mutants and genetic screens, purification of intracellular compartments, in vitro reconstitution, in vivo protein interactio, immunoprecipitation, immunofluorescence, qPCR, RNAseq, ChIPseq, flow cytometry

During these practicals 2 main experiments are proposed: Transfection and Intracellular pH assessment

1. Transfection: Eucaryote HeLa cells are cultivated for this experiment. A transitory transfection is done using plasmids that allow fluorescence localisation in the cell using targeted proteins or organelles. Transfected cells are observed and analysed using fluorescence microscopy. Images are aquired and interpreted.
2. Intracellular pH: Intracellular pH is assessed using yeast expressing a pH sensitive fluorescent protein (pHluorin), targeted either in the cytosol or in the mitochondria. In addition, a membrane permeant fluorophore is used to measure intravacuolar pH. A calibration curve is established. The cell responses to glucose addition is investigated, in the presence of various inhibitors. Quantitative data are recorded and analyzed to measure intracellular pH gradients, in relation to cell physiology.

Molecular biology course

Documents are accessible through Chamilo

Test

Semester 8 - The exam is given in english only 

Additional Information

Semester 8 - This course is given in english only