Dementia Research Institute at Cardiff University uses the FLIPR Penta System to study the role of microglia in Alzheimer’s disease

Dr. Emily Maguire and her team at the UK Dementia Research Institute at Cardiff University aim to identify the causes of Alzheimer's disease that will improve the care, diagnosis, and, ultimately, treatment of patients.

Research in Emily’s group focuses on identifying what’s different between the brain cells of people with and without Alzheimer’s disease. Specifically, they are examining a distinctive class of brain immune cells known as microglia, which have emerged as crucial contributors to the progression of Alzheimer's disease. They use genetic testing to estimate Alzheimer's risk and collect blood samples from two groups: low-risk individuals and high-risk individuals already affected by the disease. The blood cells are transformed into stem cells and then differentiated into microglia. This approach enables the team to compare microglia from high-risk and low-risk individuals and identify differences related to Alzheimer’s disease. This research would allow scientists to develop specific therapies for Alzheimer’s disease in the future.

A substantial body of prior scientific research extensively documents the malfunctioning of calcium signaling in cells affected by Alzheimer's disease. Dr. Maguire says that for this reason, it was important to compare changes in calcium signaling between healthy and diseased microglia. Dr. Maguire had previously examined calcium signaling between cells with varying levels of dementia risk at the individual cell level using an inverted microscope. Although this technique provided valuable data, it proved intricate and time-consuming. Consequently, when faced with the challenge of studying calcium signaling in 60 cell lines, the FLIPR Penta offered the ideal solution.

The team utilized the FLIPR Penta to compare the response of microglia from patients with high Alzheimer's risk to those from patients with low Alzheimer's risk when exposed to known calcium modulators. These comparisons aimed to identify discrepancies in calcium signaling between diseased and non-diseased cells, potentially shedding light on the underlying pathology.

The approach involved plating cells in a 384-well plate and staining them with a green fluorescent Calcium 6 dye from Molecular Devices. After incubation, they loaded a specific compound (e.g., ATP) into another plate. Both plates were placed in the FLIPR Penta, which added the compound to the cells and recorded changes in fluorescence both before and for 5 minutes following the addition of the compound. This enabled the team to measure the calcium released upon application of the compound and, thus, the level of cell signaling. To standardize the results, the team also captured an image of the cell plate and normalized the fluorescence measurements based on the cell number in each well, enabling them to obtain accurate and reliable comparisons between wells.