Mass spectrometry (MS) measures and sorts ions based on their mass to charge ratio. In our lab we use this technology for global and targeted proteomics using a Q exactive HF orbitrap and a Sciex 6500 Q-trap respectively. Mass spectrometry can also be used to quantify small molecules e.g drug metabolites, transporter substrates and more. We routinely do this on our waters Xevo TQ MS and waters Xevo TQ-S micro.
Liquid handling robotic systems available in our lab are a Hamilton STAR and a Tecan EVO200. These are robots are primarily used for investigations of in vitro absorption, distribution, metabolism and elimination (ADME) properties of lead and candidate drug molecules within the UDOPP platform but are also available for academic research projects. These robotic systems allows mid- to high-through-put liquid handling and increase assay precision and accuracy.
Counterflow centrifugal elutriation
Counterflow centrifugal elutriation is a technique for separating cells by size, using the two opposing forces of centrifugation and buffer flow. Cells are injected into the elutriation centrifuge, and different size fractions are eluted by sequential adjustment of centrifugation speed and flow rate. In our lab, we use this technique to study different subpopulations of human liver cells.
Cell culture facility
To handle primary cells, tissues, and cell lines, our lab is well equipped with the following instruments:
- LAF-benches, CO2-incubators, liquid N2 tanks, and ultra low temperature freezers;
- Cell imaging with phase-contrast microscopy;
- Fast determination of cell count and viability by: NucleoCounter® (based on fluorescent DNA-staining), Cellometer® Vision (based on imaging cytometry coupled with bright field and fluorescence detectors), and Countess® automated cell counter;
- Trans epithelial electrical resistance (TEER) measurements.
Our clean room is specifically designed and built for cell culture and handling. In operation, it fulfills requirements of Grade B of a clean room. The air inside is constantly recirculated through high-efficiency particulate air (HEPA) filters to remove particles from the clean room. Moreover, our implemented rules and policies together with high-standard instruments enable us to maintain cell culture in antibiotic-free medium. To maintain high-quality research and minimal interference of contaminants, we monitor our cell lines on a regular basis to ensure their function and authenticity.
In Ussing chambers a specimen of human intestine is mounted between to connected chambers allowing study of permeability and transport across the intestinal barriers. In these chambers we study the permeability of small molecule drugs, metabolites, peptides, biologics and various formulations across the mucosa of the human intestine. Using human intestinal tissue allows an in vivo like assessment of interactions with different aspects of the intestinal barrier such as mucus, metabolizing enzymes and tight junctions that are not possible in other in vitro systems. Additionally, toxic and inflammatory responses of the tissue to drugs and formulations can be monitored. Through collaborations with gastroenterologists at the Akademiska hospital in Uppsala we have access to samples of human intestine from the duodenum, jejunum, and colon allowing regional studies of intestinal permeability. Changes in intestinal barrier function in health and disease of diverse patient populations is investigated.