Could you introduce yourself?

Hi, my name is Ana Svetić, and I’m a biotechnologist turned biophysicist. I’m currently in the process of finishing my PhD in Nanotechnology at the University of Trieste and Elettra-Sincrotrone Trieste. My laboratory is called Nanoinnovation, and we do lots of Atomic Force Microscopy (AFM) as a supporting lab to the synchrotron.

What is your research project about?

My research project focuses on developing a cell-derived plasma membrane model, namely Giant Plasma Membrane Vesicles (GPMVs) and their patches (GPMV patches) for correlative nanoscale imaging – imaging with AFM and correlative techniques such as epifluorescence and TIRF microscopy, to ultimately observe the membrane nanodomains.

These nanodomains are, as the name suggests, nano-sized. In cells, they are just a few nanometers, very small and transient, and there hasn’t been a technique so far that allows the visualization and formation of said domains. But these ordered domains have been observed in synthetic models, namely Supported Lipid Bilayers (SLBs) and Giant Unilamellar Vesicles (GUVs).

So, my goal is to pass from more common simple synthetic membrane models (SLB and/or GUV) to a more complex one (GPMVs), in hopes to have a better, more biologically relevant model for observation of phase separation, which is responsible for the creation of previously mentioned nanodomains.

Which France-BioImaging’s facility did you visit, and which imaging technology did you use?

I was a visiting PhD student at the Centre de Biologie Structurale (CBS) – CNRS facility in Montpellier, where I’ve used the PALM-AFM set-up as a part of my PhD period abroad.

The PALM-AFM allowed me to correlate AFM scanning with TIRF (Total Internal Reflection Fluorescence), or more specifically, have the ability to measure the fluorescence intensity of the undermost layer of the GUV patches and GPMV patches at the same time as performing an AFM scan of the same region of interest.

This ultimately allowed me to observe the phase separation and nanodomains in two ways – by seeing (TIRF) and by scanning (AFM) the surface of the membrane models.

Could you walk us through your experience accessing France-BioImaging?

I was made aware of the France-BioImaging opportunity by my mentor, Luca Costa PhD. He kindly told me about the platform, how to apply, and so I did. It was quite easy to fill out the form, and it took little-to-no-time to receive a positive reply!

I was astounded at the efficiency of the FBI and how smooth it all went, to be honest.

How did this access contribute to advancing your project?

I was already using the set-up before applying for the funding, but the funding helped me to buy and obtain AFM tips that I needed for my measurements. AFM tips that I needed were hard to get by, so I’m quite grateful for the funding of France-BioImaging. This in turn helped me get AFM scans of better quality, and at the end of the day obtain significant results for my research.

What are the next steps for your research project?

As I’ve already mentioned, I’m currently wrapping up my PhD project. Afterwards, I’m not sure, but I would like to explore more of what I’ve developed so far, and eventually localize meaningful proteins inside GPMVs and GPMV patches. This would let us know where exactly in nanodomains are proteins such as, for example, HER-2! It would be really, really cool.

What advice would you give to researchers who are considering applying for the FBI User Access Support? 

Do it! Any support is meaningful and sometimes just a little bit can change the course of your research.