EMU Científico: "Acquisition of a high-resolution flow cytometry platform equipped with a UV excitation laser"

• Grant number: 23/16895-9
• Support Opportunities: Research Infrastructure Program - Small Medium Sized
• Start date: September 01, 2024
• End date: August 31, 2027
• Field of knowledge: Biological Sciences - Biophysics - Cellular Biophysics
• Principal Investigator: Alexander Henning Ulrich
• Grantee: Alexander Henning Ulrich
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Beatriz de Oliveira Monteiro ; Bettina Malnic ; Carsten Wrenger ; Claudiana Lameu ; Daniela Sanchez Basseres ; Fábio Luis Forti ; Gerhard Wunderlich ; Marcelo Santos da Silva ; Maria Isabel Nogueira Cano ; Nicolas Carlos Hoch ; Paola Marcella Camargo Minoprio ; Patricia Castelucci

Abstract

Flow cytometry is widely used not only in the clinical environment of hospitals, but it has become a technique that is highly used and cited in indexed international journals in the most diverse fields of scientific research. In the Pubmed database (pubmed.ncbi.nlm.nih.gov) there are a total of 1,761 manuscripts published on flow cytometry in the state of São Paulo (viewed on 01/12/2023), demonstrating the high demand for the technique in the state. However, some current and strategic research areas, such as the study of exosomes, have only 120 manuscripts published by researchers from São Paulo out of a total of 24,000 internationally. This data shows that this area of research is still emerging in Brazil and is out of step with international standards. Other areas of research that depend on the identification and purification of subcellular particles are also emerging in the state. Therefore, with a view to innovating cytometry technologies, we propose the acquisition of the CytoFLEX LX - Analyzer flow cytometer Coulter. The equipment possesses high technology, a wide range of lasers and sensitivity for detecting fluorescence and nanometric particles (characteristics not found in competing instruments). This platform will allow us to carry out concurrent multiparametric analyses (up to 21 detection colors) and, above all, to characterize extracellular vesicles and other nanoparticles such as viruses, bacteria and protozoa. Thus these characteristics make the equipment a NanoCytometry platform. The equipment in this family (CytoFLEX) has high sensitivity for fluorescence due to the avalanche photodiode-type photosensors, which are more sensitive than traditional PMTs. This gain in sensitivity allows the analysis of rare and low-frequency populations. In addition, the presence of the UV laser allows the use of non-toxic DNA intercalators (excitable at 355 nm) in conjunction with other fluorophores, enabling concomitant monitoring of DNA replication and precise delineation of the S phase of the cell cycle. Because it can measure up to 21 parameters, this platform is essential for analyzing complex samples (e.g. tumor/circulatory microenvironment). Finally, it is worth mentioning that this platform is highly economical, using fewer samples and consumables per run. These advantages will benefit users and projects that aim to study physiological/immunological facets and mechanisms for potential treatments of different diseases. (AU)