Oscar Fornas

Title : Single-virus sorting by Flow Cytometry: a methodology to elucidate the virosphere

Abstract:

Viruses is the biological entity with highest biodiversity. However, we estimate that only 1% are known. Virosphere elucidation is key to the understand global microbiome. Most studies use metagenomics to find new viruses in different ecosystems, including human body, but unfortunately only allow to identify the most abundant ones. To discover nonabundant viruses, we must go for other approaches, such as single-virus genomics.

Flow cytometry is a technology that allow us to isolate particles at single-particle level and, high-resolution flow cytometry can isolate nanoparticles at single-particle level.

Nanoparticle sorting is one of the most complex applications of flow cytometry. Basically, because we are very close to or below the resolution limit of the technique. Precisely for this reason, unlike large-particle applications, we must use other technologies to validate methodology using nano-cytometry approaches. Without such validations, we cannot fully believe the data we obtain by cytometry. Huge amount of published data of nanoparticles analysis by flow cytometry generate interesting discussions but unfortunately a methodological consensus has not yet been reached.

We developed a robust and reliable methodology, using high-resolution flow cytometry, to isolate viruses at single-virus level, for subsequent single-virus genomics to reveal nonabundant viruses as a tool to elucidate global virosphere. This approach is useful study viruses at any ecosystem, from human body, oceans, to any other environment.

What will audience learn from your presentation?

The aim of this presentation is showing a new approach to isolate nanoparticles by flow cytometry. How to validate a new flow cytometry methodology of nanoparticles.

Finally, the idea is sending a message how nanoparticles must we manage by flow cytometry. This presentation will introduce the tips and tricks to deal with nanoparticles in general. Further than viruses, this can be used as an approach for all nanoparticles, such as extracellular vesicles or inorganic particles.

Biography:

Oscar Fornas, Ph.D in Biochemistry, is the Head of Flow Cytometry Unit at Centre for Genomic Regulation and Pompeu Fabra University in Barcelona, Spain, since 2001, where coordinates the assistance, supervision and training of one of the most advanced flow cytometry platforms in Europe. The Unit assists, supervise and advise more than 300 users from around 120 research groups in Spain and Europe. He supports the use of most flow cytometry applications as well as develops and implements new ones under scientific community needs.

He recently developed the Flow Karyotyping for chromosome sorting, which combined with subsequent chromosome sequencing without amplification by using Oxford Nanopore technologies becomes a new methodology to assemble structurally very complex chromosomes.

Currently, he is focused in develop strategies to detect and isolate nanoparticles by Flow Cytometry. Recently, he has developed an approach for Single-Virus sorting for subsequent single-virus genomics. This methodology allows the possibility to explore the diversity of viruses in different ecosystems and to elucidate the global microbiome, in special, the global virosphere. He has also developed a new approach to identify and isolate Extracellular Vesicles (smaller than 100nm) by Flow Cytometry, to study cell communication and, based on Extracellular Vesicles’ cargo transfer, looking for biomarkers related with diseases. His Unit is becoming a reference site for advanced Flow Cytometry applications and sorting at the single-particle level. Oscar also participates, as membership, in alliances of excellence, as Core4life, an ”Excellence Alliance of Life Science Core Facilities in Europe”.