EMU: Acquisition of ImageXpress Nano equipment for analysis and visualization of cellular multiparameters

• Grant number: 23/17262-0
• Support Opportunities: Research Infrastructure Program - Small Medium Sized
• Start date: September 01, 2024
• End date: August 31, 2027
• Field of knowledge: Biological Sciences - Pharmacology - General Pharmacology
• Principal Investigator: Valeria Cristina Sandrim
• Grantee: Valeria Cristina Sandrim
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated researchers: Arielle Cristina Arena ; Carlos Alan Candido Dias Junior ; Erick José Ramo da Silva ; Fátima Pereira de Souza ; Ícaro Putinhon Caruso ; Luiz Gustavo de Almeida Chuffa

Abstract

Live-cell image visualization is an effective tool that empowers scientists to study living organisms instantly. Especially when used in conjunction with fluorescence microscopy, this technique enables the observation of cellular components with great sensitivity and precision. By illuminating the dynamics of different compounds and structures in living cells, it is possible to clarify their biological interactions and associated biochemical activities. These assays are particularly useful when aiming for refinement in obtaining results. The ImageXpress Nano Automated Imaging System features technical capabilities suitable for these assays, providing highly sensitive and reliable results as it can operate in both 2D and 3D imaging fields, as well as time-lapse analyses. In the 3D realm, ImageXpress has the benefit of accessing a high range of human tissue characteristics, concerning cellular architecture and cellular interactions (between different types and in the intra-cell environment), as well as the physiological aspects, such as diffusion and metabolism - an essential feature in pharmacology. The incorporation of these assays, especially 3D analysis, amplifies research possibilities once it fills the lack of knowledge among simple cell cultures and animal models. Another crucial point is the need to precisely quantify the number of cells in a biological experiment. Assays related to drug safety and efficacy, cell division, and cell senescence depend on evaluating the number or density of cells in a specific area. Additionally, the analysis of cell behavior, such as migration, is frequently conducted in vitro to unravel the underlying cell program and physiology. The evaluation of cell migration can be executed in a pre-defined environment and for a short period, utilizing live time-lapse images of cells. These techniques are useful to compare different substances' effects. Currently, available assays often require several hours of manual analysis, increasing the likelihood of sampling errors and reducing study refinement. The broad applicability of this equipment aligns with ongoing research at the Institute of Biosciences at São Paulo State University, potentially enhancing expected results. Through fluorescent labeling, the equipment can identify proteins in various cellular compartments, whether fused or isolated, as well as their trafficking within the cell, essential for examining the behavior of certain drugs and the cascades of reactions triggered after the binding of these compounds to their receptors or other cellular components. One target protein under study, for instance, is EPPIN, a pharmacological target that may contribute to the development of male contraceptives based on inhibiting sperm function. Furthermore, the evaluation of proton pump inhibitor effects on blood pressure and nitric oxide bioavailability in gestational hypertension can also be measured via ImageXpress. These aspects are related to inflammatory pathways altered in various diseases and syndromes. Studying these pathways will contribute to advancing the understanding of complex diseases, such as cancer, and even fundamental aspects of psychiatry, such as the regulation played by the brain-gut axis. Despite their broad nature, all these pathways have correlated foundations and involve ongoing local research. They encompass the study of cellular environments and the intimate interaction of their structural compounds or messenger cascades, making the use of sensitive tools crucial for obtaining accurate and innovative data. (AU)