Nuclear translocation is a critical cellular mechanism for the function and regulation of eukaryotic cells. In this issue of TekTalk we’ve compiled a variety of resources that highlight automated image analysis techniques for quantifying nuclear translocation events within cell model systems, including tips and best practices for optimizing translocation assays. In the featured application notes below, the Agilent BioTek automated imaging systems and Gen5 image analysis software are employed to monitor changes in intracellular protein location using both endpoint and kinetic live cell applications. The use of nuclear stains and powerful Gen5 dual masking capabilities enable accurate determination of nuclear to cytoplasmic ratios of proteins of interest. The automated microplate-based format supports efficient, high-throughput characterization of treatment response and screening applications.
As critical components of the innate immune system, macrophages respond to microbes by recognizing molecules, such as the gram-negative bacteria product lipopolysaccharide (LPS), via Toll-like receptors. Receptor activation stimulates a complex signaling network that involves, among others, the NF-κB pathway. This application note quantitates the nuclear translocation of the protein RelA, a component of the NF-κB transcription factor complex, in macrophages using image-based analysis.
Hepatocarcinoma is one of the most common cancer types in the U.S. with limited treatment options and poor survival rate. Recently, cancer bioenergetics emerged as a promising source of targets to develop novel anticancer agents. The identification of relevant biomarkers is essential for screening and validation of small molecules proposed as chemotherapeutic agents. In this regard, there is a great need for high-throughput/high-content methods, which require short, reproducible sample preparation techniques.
BioTek Cytation 5 is a modular, upgradable multimode reader that combines automated digital microscopy and conventional microplate detection. Cytation 5 includes filter- and monochromator-based detection; the microscopy module provides up to 60x magnification in fluorescence, brightfield, color brightfield and phase contrast. Incubation to 65 °C, shaking and Gen5 software are standard.
Watch this video to learn more about the complete Cytation line of cell imaging multimode readers.
RAW264.7 G9 cells require careful handling prior experimental use. They are expanded in non-treated tissue culture flasks such as P/N 15800 T-75 flasks from Nunc. The use of digestive mixtures such as trypsin should be avoided as they will result in cellular activation and a cessation of growth. Cells are detached by adding cold 2 mM EDTA in PBS for 5 minutes.
Wednesday, December 07, 2022 | 12 PM EDT
Speakers: Jamie Stern, Ph.D., Field Applications Scientist, Cell Analysis, Amanda Herberger, PhD, Field Applications Scientist, Cell Analysis, Cody Narciso, Field Applications Scientist, Cell Analysis, Agilent
Over the last seven decades, spheroids have emerged as an attractive model for many scientific fields, including drug development and disease progression. Though spheroids have been in use for an extended period, researchers often have questions about how to take advantage of this 3D system. In this webinar, we will discuss many factors that underlie a successful spheroid experiment, including the considerations that must be given to sample preparation, data capture, and analysis.
Topics will include:
Sourcing your microplates shouldn't be an added task in preparing your cell-based workflows. Agilent now offers cell culture and imaging microplates so you can complete your workflows seamlessly.
Expect the quality you need with TC surface treatment enhancing the cell attachment while the microplate lid which accompanies every plate maintains a sterile growth environment with a low risk of cross contamination.
Buy an Agilent BioTek MultiFlo FX multimode dispenser with a single peristaltic pump (MFXP1 or MFXP2 models) and a secondary peristaltic pump (part number 7210010), and receive a 50% promotion discount on the secondary peristaltic pump.
Buy an Agilent BioTek Cytation C10 confocal imaging reader containing the “M” module (monochromator-based UV-Vis absorbance, fluorescence, and luminescence), and receive the instrument for the same price as the Cytation C10 without the "M" module.