Project Work

During this course the students perform 4 weeks of project work regarding ongoing work at the laboratory by doctoral and postdoctoral researchers. The specific projects offered depends on what work is ongoing in the laboratory at the time of the course. In 2016 these projects included “determining how omega-3 fatty acids effects the expression of fatty acid elongases”, “measuring how selective β2-agonists effects glucose uptake in skeletal muscles” and “using qPCR to determine how different sera effect adipogenic gene expression”.

Lab Stations

The students of the course get the opportunity to perform three one-day lab stations together with doctoral and postdoctoral researchers. In these lab stations the students of the course get to learn how to use different common methods performed in the study of molecular physiology.

Mitochondrial Oxygen Consumption

Mitochondria utilize oxygen in order to produce ATP, the main energy currency of the cell. The oxygen consumption can be measured by an oxygraph and be used to calculate mitochondrial activity. In this lab the students learn to isolate mitochondria from tissue and to measure oxygen consumption in a variety of conditions by adding substrates, inhibitors and uncouplers.

Thin Layer Chromatography

Thin Layer Chromatography (TLC) is a commonly used method for separating and analysing compounds. It operates by compounds having distinct retention between a stationary phase and a mobile phase while capillary forces drives the mobile phase upwards along the stationary phase. In this lab the students learn to use TLC to identify and distinguish between different fatty acids.

qPCR

Quantitative Polymerase Chain Reaction (qPCR) is a common method for measuring the abundance of specific RNA. It utilizes a chain reaction of polymerase to mass copy a string of DNA (made from RNA with reverse transcriptase) while measuring the quantity by changes in fluorescence from SYBR Green. In this lab the students learn to use qPCR to measure the abundance of mRNA in a sample.

Western Blot

Western blot is a common method for protein identification and quantification. It utilizes an electric gradient to separate protein based on size in the presence of SDS. Antibodies, conjugated to a reporter causing light emission, are used to detect and quantify a specific protein. In this lab the students learn to use western blot to quantify UCP1 protein in a mice model exhibiting Cushing’s syndrome.

Glucose Uptake

Glucose uptake is an important part of energy regulation in cells and regulates blood glucose levels. Glucose uptake is measured by utilizing radioactively labelled unmetabolizable glucose and later quantifying the radioactive emissions. In this lab the students learn how to measure and quantify the glucose uptake in cultured skeletal muscle cells.

MRI and Metabolic Chambers

Magnetic Resonance Imaging (MRI) utilizes the absorption and re-emission of radio waves in molecules under the influence of a strong magnetic field to calculate fat and lean mass. The metabolic chambers measure the oxygen consumption in order to measure metabolism. In this lab station the students get the opportunity to measure fat and lean mass as well as metabolism.

Immunocytochemistry

Immunocytochemistry is a method for visualizing components of cultured cells. It utilizes fluorescent dyes, which bind to a specific target, or fluorescently marked specific antibodies and a fluorescent microscope. In this lab stations the students get the opportunity to learn how to mark and visualize cultured brown adipocytes’ nuclei and lipid droplets in a microscope using both dyes and antibodies.

Cell Culture and Sterile Techniques

Cell cultures are commonly used and are the basis for many experiments (including many of the other lab stations). Animal cell cultures are susceptible to infection, as they have no immune system, so sterile techniques are of paramount importance. In this lab station the students learn how to culture and care for one of the most used immortalized cell cultures for studying adipose development 3T3-L1.

Flow Cytometry

Flow cytometry is a method of quantifying cell types present in a solution. It uses fluorescently tagged antibodies, binding to specific surface receptors, on different cell types to quantify them while the cells flow past a detector. In this lab station the students learn how to use flow cytometry to quantify B and T lymphocytes in mice spleen taken from mice housed at different ambient temperatures.