QualityNano funded visit to UCD – November/December 2013
The visit to the Centre for BioNano Interactions through the QualityNano Transnational Access application was an excellent opportunity to broaden my skills in the field of protein chemistry and cell biology. During my 15-day visit, I renewed my knowledge on cell/tissue culture work, brief characterisation of nanoparticle suspensions, fluorescence staining, etc. I didn’t have any experience in immunostaining, confocal microscopy and immunoblotting when I started my QualityNano TA visit. The main aim of my project was to observe lysosomal damage in relation to altered expression and/or distribution of chloride channels on lysosomal membranes in A549cells. Even though the chosen antibody for the chloride channels was observed to be unspecific and therefore I didn’t get the expected results, I learned the protocols for both immunostaining and immunoblotting and all that work I can continue at my home institution at University of Ljubljana, Slovenia. I was also skilled in using epifluorescence and confocal microscopes in self-dependant manner and the output were, besides my experience, are also some confocal micrographs (below).
A549 24 h Control
Swollen lysosomes (Lamp-1/488)
QualityNano funded visit to UCD – March/April 2014
Thanks to a QualityNano grant I was able to undertake a visit to the Centre for BioNano Interactions (CBNI) at University College Dublin for two weeks in March 2014, under the supervision of Dr. Marco Monopoli. The aim of this visit was to make use of the expertise and equipment at CBNI to improve our understanding of the agglomeration state of dextran coated ultrafine superparamagnetic iron oxide nanoparticles (dUSPION) under our selected in vitro experimental conditions, in addition to training in hard protein corona analysis. The agglomeration analysis was performed using differential centrifugation sedimentation (DCS) using both Fe2O3 and Fe3O4 dUSPION in cell culture medium with both 1% and 10% foetal bovine calf serum; both particle types under the same conditions were also used for hard protein corona analysis by SDS PAGE. A substantial amount of data was generated using the DCS technique allowing comparison with dynamic light scattering (DLS) data previously generated at our home institution (Swansea). The training received on protein corona analysis also proved to be invaluable as this was a technique our group had no previous experience of, despite having the equipment available.In addition to the generation of important data, I have been able to carry out this technique at Swansea on my return and train others in the process.
I very much enjoyed my time at UCD and greatly appreciated the opportunity to work with a research group that is so experienced in bio-nano interaction. During the visit I was given the opportunity to present a seminar of my work to date and explain what I hoped to gain while at UCD and was provided with valuable feedback and suggestions. All data produced as a result of my QualityNano TA will make an excellent contribution to my PhD thesis. Additionally, I aim to present the work at future conferences and incorporate it in any publications that result from my project. I am very grateful to QualityNano for funding this trip and to Dr. Monopoli and the rest of the CBNI group for hosting me and would very much like the opportunity to work with them in future.