"EMU: Acquisition of a Confocal Laser Scanning Microscope Equipment for structural and functional analyzes of cells (prokaryotes, eukaryotes), tissues (humans, animals, plants), organoids, biofilms and materials (biomimetics)"

• Grant number: 23/17123-0
• Support Opportunities: Research Infrastructure Program - Small Medium Sized
• Start date: September 01, 2024
• End date: August 31, 2027
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Marlise Inêz Klein Furlan
• Grantee: Marlise Inêz Klein Furlan
• Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil
• Associated researchers: Renata de Oliveira Mattos Graner ; Ricardo Della Coletta ; Valentim Adelino Ricardo Barão

Abstract

The proposal aims to acquire an EMU-Confocal Laser Scanning Microscope for structural and functional analyses of cells (prokaryotes, eukaryotes), tissues (humans, animals, plants), organoids, biofilms, and materials (biomimetics). A confocal microscope is an excellent tool for blocking out-of-focus light and thus performing optical cutting through the sample. This procedure allows fluorescence to be quantified with very high spatial precision. It is possible to observe and quantitatively analyze: 1. tissue structures (animals, plants, invertebrates) in vitro and in vivo, organoids grown in laboratories, free microorganisms or living as biofilms (construction of 3D images); 2. cells (eukaryotes and prokaryotes), how the cells are organized among themselves and how they are connected to each other, which occurs via direct contact (adhesins, specific junctions) or extracellular matrix components (tissues and microbial biofilms ); 3. extracellular matrix (discrimination of constituents in tissues, organoids, benign or malignant tumors, microbial biofilms); 4. dynamics of microenvironment formation within organoids or biofilms, e.g. for metabolic (functional) assessment; and other applications. However, to generate meaningful data via confocal microscopy, it is essential to consider aspects of quantitative image acquisition, including optimizing sample preparation, choosing the most suitable microscope for a given application, and configuring its parameters. Therefore, the EMU-confocal microscope was requested to meet the growing demands at the Microscopy and Imaging Center (C.M.I.) at FOP-UNICAMP, which has a team with expertise in various microscopies. There is a confocal microscope at C.M.I., but it has reached obsolescence, and the supplier does not guarantee the supply of parts or its maintenance, in addition to its configuration not meeting the necessary requirements for the analysis of many samples from projects underway at the Institution and from researchers external parties who seek out C.M.I.. The host institution is a hub for training highly qualified human resources and generating knowledge that impacts global scientific production. In Piracicaba-SP, in addition to the outdated confocal microscope at C.M.I., there are two other confocal microscopes at the "Luiz de Queiroz" School of Agriculture (ESALQ)-University of São Paulo (USP). However, they are also outdated and have not satisfactorily met several requirements for plant sample analysis. Thus, this lag in the confocal microscope research tool negatively impacts the development of research at the frontier of knowledge in different areas and institutions. The EMU to be purchased has a configuration that will meet current and future demands, as it possesses: 1. 4 lasers [405 (UV), 488 (blue), 561 (green), 638 (red)], 2. latest detection system silicon-based SilVIR" generation that promotes accurate and reproducible data acquisition by combining two advanced technologies - a silicon photomultiplier (SiPM) and a patented fast signal processing design (provides high dynamic range/amplitude, low noise, without sensitivity degradation and less sensitivity variation compared to other detectors); 3. TruSpectral technology combined with a highly sensitive SilVIR detector, which allows the use of up to six channels simultaneously, among other specifications. Therefore, the requested EMU will assist in the execution of projects using different types of samples for analysis of mechanistic and applied studies (structural and functional) that will contribute to the advancement of knowledge, which will be reverted to benefits for society. The C.M.I. is already a reference center for microscopy in São Paulo, and the acquisition of the EMU will help maintain its relevance in the scientific community. (AU)