Agilent BioTek Lionheart FX Automated Microscope

The ability to manipulate and monitor microbial organisms, such as bacteria and yeasts, is critical to exploiting their many applications, from use in medical products and biomedical research to environmental remediation and food production. Meeting the increasing demands of microbial monitoring is therefore of vital importance, requiring laboratories to optimize their workflows with the most accurate, reliable and up-to-date instrumentation.

Download this application guide to learn how to take your bacterial monitoring to the next level and achieve real-time results, increased productivity and greater efficiency with the latest monitoring technologies, as we cover:

• The principles of light scattering techniques
• New ways to enhance the performance of brewing yeasts
• The advantages of brightfield microscopy for monitoring in real time
• How to achieve optimum productivity, without multiple instruments, whilst reducing costs

With BioTek’s Lionheart FX Automated Microscope and Gen5 Image Analysis Software, total cells/volume can be calculated for almost any type of cell, using digitally captured and analyzed images, independent of probes or stains. This process is demonstrated in this free infographic that shows how to quantify pre- and post-fermented yeast cells in corn mash.

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. Here we quantitate the nuclear translocation of the protein RelA, a component of the NF-κB transcription factor complex, in macrophages using image-based analysis.

In this application note, BioTek demonstrates high-throughput screening and analysis of zebrafish embryos treated with several developmental toxins. Here, BioTek exposes embryos to ethanol, retinoid acid, and cyclopamine, then capture brightfield images of the embryos in multi-well plates with Lionheart™ Automated Microscope. The images are analyzed for size and shape using Gen5™ software. The combination of high-throughput imaging and analysis using a vertebrate model makes this an attractive method for toxicology screening.

Autophagy is critical for the maintenance of cellular homeostasis. However, dysregulated autophagy can lead to death of healthy cells and survival of cancerous cells. Here we describe the use of CYTO-ID® Autophagy Detection Kit in combination with automatedobject-based spot counting to quantitatively assess the effects of starvation and rapamycin on cellular autophagy by determining the size and number of autophagosomes per cell.

GPCR activation was kinetically monitored using a Ca2+ biosensor that was transfected along with the human muscarinic M1 receptor into HEK 293 cells. Rapid Ca2+ flux was evident upon carbachol stimulation that peaked 1 second after injection, followed by a decay back to baseline fluorescence over 90 seconds. Imaging also allowed for the monitoring of single cells within the cell population in the field of view where differences in kinetics can be assessed. Suitable assay performance was achieved using a % Responders readout (% of cells responding to carbachol stimulation) that provided z’ values above 0.5 and consistent pharmacology.

This poster describes an automated imaging-based approach to characterize the IP3 /DAG signaling pathway using multiplexed R-GECO Ca2+ and DAG fluorescent biosensors from Montana Molecular. This method produces detailed kinetic profiles of Ca2+ flux and DAG levels following activation of Gq-coupled receptors within the same population of cells. Background subtraction and image analysis tools enable detection of changes in Ca2+ and DAG levels that are more sensitive than techniques relying on bulk fluorescence measurements. Additionally, this method enables single-cell analysis for detailed characterization of response dynamics and subcellular effects.

Monitoring calcium flux is an important method for characterizing GPCR (G-protein coupled receptor) activation. This application note describes a live cell imaging based approach to quantify Ca2+ flux kinetics using the Lionheart™ FX and Fluo-4 Ca2+ indicator dye that delivers sub-second resolution and a large assay window. This method enabled continuous monitoring of intracellular calcium, provided very good sensitivity and generated detailed kinetic profiles for GPCRs.

In this application note describes the use of the Lionheart™ FX Automated Live Cell Imager and the ONIX2 Microfluidic Platform to monitor yeast cell growth through the analysis of kinetic brightfield images of yeast cells under a variety of conditions in real time. The yeast under inspection is the yeast Saccharomyces cerevisiae. Strains of the yeast Saccharomyces cerevisiea serve critical roles in the production of many different products including food staples, medicines, and biofuels.

This application note describes using the ONIX2 System in conjunction with the Lionheart™ FX Imager to rapidly image and analyze perfused tissue culture cells in multiple fluorescent colors and brightfield.

Chris Laucius, Ph.D., Imaging and Microscopy Specialist at BioTek Instruments, explains how the plate washers and imagers from BioTek Instruments can help with every step of your workflow: from gentle media exchange, sample prep, image acquisition all the way to data analysis.

Scientists at Stony Brook University, New York, have discovered that the previously known antiviral protein BST2, also known as tetherin or CD371, functions as an oncogenic driver in breast cancer-promoting cell adhesion, growth, migration and invasion of tumor cells. In this video, discover how the automated imaging system of BioTek's LionheartTM FX platform has enabled the group to unearth many of the structure-function relationships of BST2 in cancer, whilst eliminating investigator bias and enabling highly accurate acquisition of kinetic data. Read our article for more information on research in the Okeoma lab.

Dr Arne Giebeler, Sales Support Specialist at BioTek Instruments Inc., describes how the compact Lionheart™ FX has enhanced environmental and focusing controls to facilitate multiple cell imaging applications.

Watch video highlights from the Neuroscience 2018 annual meeting in San Diego, from the latest research presented, to the new technologies innovating neurobiology research

How advanced imaging technology is progressing drug discovery through understanding of GPCR-dependent second-messenger systems

A laboratory in the US is using advanced imaging to decipher the role of a ‘double-agent’ protein in breast cancer, and to help develop drugs targeted at disrupting its function

Find out how experts have been using this new imaging technology and read their verdicts on its performance

Learn how a lab in the US has perfected a high-throughput assay in mouse organoids for testing potential new cystic fibrosis drugs