Cell based assays: The optimal reads function for fluorescence bottom measurements

19 Feb 2024

In this application note, Tecan highlights the Optimal Read (OR) function for bottom reading fluorescence intensity measurements in adherent cell lines. Performing single fluorescence reads in the center of the well can give a false impression of the cell density in cases where the cells show non-uniform distribution across the growth surface. The OR function for fluorescence bottom measurements is designed to enable uniform well illumination during excitation with whole-well emission, ensuring that even heterogeneous cell layers can be reliably analyzed. The note provides detailed information on the experimental setup, including measurement settings, and presents results showing improved uniformity and reproducibility of well-to-well measurements.

Links

Tags

Fluorescence SpectroscopyFluorometers and spectrofluorometers (also called fluorescence spectrometers) are used to measure the intensity and wavelength of fluorescent light emitted from a sample after excitation by illumination. Spectrofluorometers utilize monochromators to select the desired wavelengths, whereas filter fluorometers employ a set of filters. Spectrofluorometers for measuring steady-state fluorescence and lifetime fluorescence (or time-resolved fluorescence) are available, as well as fluorescence microscopes and microplate readers. Find the best fluorescence spectroscopy products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell Lines Stem Cells and Primary CellsPrimary cell cultures, established cell lines and stem cells are vital for <i>in vitro</i> and <i>ex vivo</i> experimentation. High-quality cells, optimized for your applications, alongside optimized cell substrates, growth medium and supplements, are critical for experimental success. Explore a range of cells suitable for your applications, including isogenic cell lines, competent cells, induced pluripotent stem cell (iPSC)-derived cell lines, fungal/bacterial/mammalian cell lines, stem cells and cancer cell lines. Find the best cells for your research in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.FluorescenceThe emission of fluorescence occurs when a photon of energy is supplied to a fluorescent chemical compound by an external source, causing it to become excited. Fluorescence can be detected and measured for different purposes using microplate readers, fluorescence microscopes, fluorescence scanners, and flow cytometers.Cell ImagingCell imaging can be achieved using a number of techniques including confocal microscopy, transmission electron microscopy, atomic force microscopy, and light sheet microscopy.Lead OptimizationLead optimization is a phase in the drug discovery process. A lead is a compound that has demonstrated desired biological effects on a target. Optimization involves making chemical modifications to the structure in order to generate analogues of the original compound, which have higher potency and safety profiles.
Cell based assays: The optimal reads function for fluorescence bottom measurements