Real-Time Cell Insights with Agilent xCELLigence RTCA eSight

A Joe Blogs post by Joe Roberts, PhD

Cells don’t stand still, so why should your assays? The Agilent xCELLigence RTCA eSight™ goes beyond snapshots, revealing real-time cell behaviour with label-free impedance and live imaging.

By pairing biosensor impedance technology with live-cell imaging, eSight enables researchers to monitor cell health, function, and behaviour simultaneously, continuously, and in real time. It’s not just more data, it’s deeper, more meaningful insight into what your cells are really doing.

Agilent xCELLigence RTCA eSight real-time cell analysis platform in incubator

How Real Time Cell Analysis (RTCA) Technology Works

At the heart of RTCA eSight are proprietary E-Plates, embedded with gold biosensors. These electrodes non-invasively measure impedance, which reflects cell metrics such as:

  • Proliferation
  • Adhesion strength
  • Morphological changes
  • Migration and differentiation

Because impedance is recorded continuously, you capture events as they happen, in seconds, minutes, hours, or days, without disturbing the cells. The temporal resolution is exquisite, allowing you to see subtle shifts long before they’re visible under a microscope.

Agilent xCELLigence RTCA E-Plate with integrated gold biosensors for real-time, label-free impedance-based cell analysis.

Multi-Modal Cell Analysis: Impedance Meets Live-Cell Imaging

In concert with impedance, eSight’s imaging module provides brightfield plus three fluorescence channels (red, green, blue). This lets you visualise your cells directly while validating and enriching the kinetic impedance data.

The result is a spatial and temporal view of cell populations at an unprecedented level of detail, ideal for assays like proliferation, cytotoxicity, and apoptosis. And importantly, impedance and imaging are performed on the same cells, not replicate wells, so your data are directly correlated.

Kinetic comparison of apoptosis readouts using Agilent xCELLigence RTCA eSight showing cell index and fluorescence markers after drug treatment. Live-cell imaging of apoptosis markers with Agilent xCELLigence RTCA eSight, showing Annexin V, Caspase 3, and nuclear BFP fluorescence with matching reagents.
Left: Live-cell imaging of apoptosis markers with Agilent xCELLigence RTCA eSight, showing Annexin V, Caspase 3, and nuclear BFP fluorescence with matching reagents. Right: Live-cell imaging of apoptosis markers with Agilent xCELLigence RTCA eSight, showing Annexin V, Caspase 3, and nuclear BFP fluorescence with matching reagents.

Streamlined RTCA Workflow

One setup, two data streams. Here’s how simple it is:

  1. Seed your cells into an E-Plate.
  2. Insert the plate into eSight (inside your incubator).
  3. Define your assay in the RTCA software.

From there, impedance and imaging data are captured automatically, then integrated into a single timeline for straightforward analysis. The software even supports outputs like:

  • RTCA images
  • KT50 (time to 50% cytolysis at a given effector-to-target ratio)
  • % Cytolysis dose response curves
  • IC50 calculations

No juggling between platforms – just a unified dataset ready for export.

Applications Across the Board

Researchers are already applying RTCA eSight to:

  • Immune-cell killing assays – track cytolysis in real time with kinetic precision.
  • Virology – Screening and characterizing antiviral drugs in real time
  • Proliferation and apoptosis studies – capture early events, validate with imaging, and overlay with fluorescent markers.
  • Cell heterogeneity analysis – reveal subpopulation responses that would be lost in averaged data.

Joe’s Takeaway

The Agilent xCELLigence RTCA eSight is more than the sum of its parts. By integrating patented biosensor impedance with live-cell imaging, it delivers:

  • Continuous, label-free monitoring of cell health and function
  • Brightfield and Three-colour live-cell imaging for direct visual validation
  • Unified kinetic datasets from the same cell population
  • Broad versatility across immuno-oncology, virology, and general cell biology

For researchers who want to move beyond static snapshots, eSight offers a powerful new way to see biology unfold in real time, with the reproducibility, temporal resolution, and ease-of-use that modern labs demand.

If you’re interested in a demo of the eSight, contact us today!

Until next time… happy experimenting!

Joe Blogs

Meet EYRA: Multiplexing Reimagined

A Joe Blogs post by Joe Roberts, PhD

When it comes to multiplex protein analysis, researchers need accuracy, speed, and simplicity. Traditional flow-based systems have long been the standard, but they bring challenges: sheath fluids, blocked probes, and constant maintenance. Enter the Mabtech EYRA™ – a fluidics-free multiplex immunoassay platform that reimagines how scientists generate cytokine and biomarker data. With confocal imaging, RAWsphere analysis, and compatibility with EYRAplex bead kits, EYRA makes multiplexing faster, simpler, and more reliable.

Mabtech EYRA is now available in Australia and New Zealand

 

Multiplex Without Compromise

With EYRAplex magnetic bead assays, EYRA can quantify more than 30 analytes from a single sample – whether that’s serum, plasma, or cell culture supernatant. This means less sample consumption, fewer runs, and richer datasets for every experiment.EYRAplex assay schematic

At the core of EYRA’s precision is the RAWsphere image analysis algorithm. It identifies each bead, links it to the correct analyte, and quantifies the PE signal with high resolution. Whether you’re measuring six cytokines or a full 30+ panel, RAWsphere ensures your multiplex data is accurate and reproducible.

Watch video to see how EYRA works

Fluidics-Free Technology

EYRA is built around a completely flow-free design. No sheath fluid, priming, waste handling or blocked probes.

Instead of pushing samples through fluidics, EYRA uses confocal microscopy to image settled magnetic beads directly in the wells of a 96-well plate. Each bead carries a unique fluorescent dye signature for identification, while analyte-bound PE-labelled antibodies provide the quantitative signal.

The result? You insert your plate, select your assay in the intuitive Mabtech Opal™ software, and hit read. Fifteen minutes later, you have fully processed data – with your samples never leaving the wells.

From Plate to Excel - Fast

Opal™ software comes preloaded with templates for every EYRAplex kit. Standards, plate layouts, and gating are handled automatically – so you spend less time setting up and more time on results.

Once the plate read is complete, results are exported directly into Excel, with options for bulk export if you’re working on large studies or multiple plates. No manual reformatting. No data wrangling headaches.

The Maintenance-Free Mindset

Because EYRA is fluidics-free, there’s no daily calibration or cleaning. No flushing, no wasted consumables, and no time lost to instrument downtime. It’s genuinely a plug-and-play experience – switch it on, run your plate, and walk away with your data.

Joe’s Takeaway

The Mabtech EYRA isn’t just another multiplex platform – it’s multiplexing reimagined. By removing fluidics and harnessing high-resolution confocal imaging, EYRA delivers:

  • High-plex capacity: 30+ analytes per well
  • Fast turnaround: ~15 minutes per plate
  • No maintenance: no daily cleaning or calibration
  • Accurate, reproducible results: powered by RAWsphere analysis

For labs looking to streamline their workflow without sacrificing data quality, EYRA offers a fresh, frustration-free alternative to traditional flow-based systems.

Until next time… happy experimenting!

Joe Roberts, PhD
Product Manager
Millennium Science

Joe Blogs


Spot the Difference: ELISA vs ELISpot vs FluoroSpot

A Joe Blogs post by Joe Roberts, PhD

When it comes to measuring proteins such as cytokines, antibodies, or growth factors, there’s no shortage of options. ELISA, ELISpot, and FluoroSpot all have important roles to play. While each assay relies on antibody-based detection, knowing which one to use (and when) can mean the difference between simply collecting data and gaining real insight.

As a Product Manager at Millennium Science, I speak with researchers every day who are deciding between these techniques. In this blog we will explore each assay to help you spot the difference and help you choose the right assay for your next experiment.

Comparison at a Glance – ELISA vs ELISpot vs FluoroSpot

ELISA vs ELISpot vs FluoroSpot comparison table

ELISA – The Trusted Workhorse 📊

Best for: Quantifying total soluble protein in biological fluids.

ELISA (Enzyme-Linked Immunosorbent Assay) is one of the most widely used immunoassays for detecting and quantifying soluble proteins, such as cytokines, antibodies, and hormones, in serum, plasma, or cell culture supernatants. It’s robust, scalable, and ideal for high-throughput analysis when you need accurate, reproducible concentration data.

How it works (Sandwich ELISA):
A capture antibody is coated onto a high-binding plate and binds the target protein in your sample. A biotinylated detection antibody binds a different epitope, followed by a streptavidin–enzyme conjugate. Addition of a colourimetric substrate produces a measurable signal proportional to protein concentration.

Why researchers choose it:

  • Delivers quantitative results (e.g., pg/ml or ng/ml)
  • Compatible with high-throughput and automation
  • Well-established, with widely available equipment
  • Ideal for comparative analysis across multiple samples

Limitations:

  • No information on the number or type of cells producing the protein
  • Sensitivity can be lower than single-cell assays
More information

ELISpot – Spotlight on Secreting cells

Best for: Counting individual protein-secreting cells.

ELISpot (Enzyme-Linked ImmunoSpot) detects and counts cells that secrete a specific protein. It’s particularly valuable when studying immune responses where the frequency of antigen-specific T or B cells is low – for example in vaccine research, oncology, and autoimmune disease studies.

How it works:
Live immune cells are added to a PVDF plate pre-coated with a capture antibody. When stimulated, the cells secrete the target protein, which is immediately bound near the cell. After washing away cells, a detection antibody and enzyme conjugate are added. A precipitating substrate forms a visible spot at each secretion site – each spot representing a single responding cell.

Why it stands out:

  • Extremely sensitive – can detect one responder cell in >100,000
  • Functional readout of immune activity
  • Ideal for rare antigen-specific responses
  • Widely used in T-cell response monitoring, vaccine trials, allergy research, and immuno-oncology

Pairing with Mabtech’s ASTOR2 automated ELISpot reader delivers rapid, high-precision spot counting with consistent, reproducible results across entire assay plates, and when used with Mabtech monoclonal antibody pairs and ready-to-use ELISpot kits, ensures reliable performance across species and sample types.

More information

FluoroSpot – Multiplex Made Easy 🌈

Best for: Analysing polyfunctional immune responses.

FluoroSpot builds on the ELISpot principle but uses fluorescently labelled detection antibodies to identify multiple proteins secreted by the same cell – typically 2-4 proteins. This allows you to measure not just whether a cell responds, but how it responds.

Why it’s powerful:

  • Detects polyfunctionality – cells secreting multiple cytokines
  • High sensitivity with low background
  • Enables simultaneous detection of different functional subsets
  • Excellent for vaccine research, infectious disease, and cancer immunotherapy

Pairing with Mabtech’s IRIS2 ELISpot, FluoroSpot and FociSpot reader enables precise, high-resolution spot detection across multiple fluorescence channels.

More information

Joe’s Takeaway

  • ELISA → When you want to know how much protein is present in a sample.
  • ELISpot → When you need to know how many cells are producing a protein.
  • FluoroSpot → When you need to know which protein a cell is producing, and whether it’s producing more than one at the same time.

ELISA vs ELISpot vs FluoroSpot diagram

In many projects, using ELISA and ELISpot/FluoroSpot together can give the clearest picture — combining quantitative bulk measurements with functional single-cell insights.

If you’d like help selecting the right assay for your research, get in touch with our team at Millennium Science. We’re local experts, and Mabtech’s range covers everything from research use to clinical-grade applications.

Until next time… happy experimenting!

Joe Roberts, PhD
Product Manager
Millennium Science

Joe Blogs

 

 

Welcome to Joe Blogs

A Fortnightly Take on Life Science Technology 🔬

Hi, I’m Joe Roberts, PhD – Product Manager here at Millennium Science, immunologist at heart, and someone who’s passionate about helping researchers get the most out of their tools, assays and experimental design. My career has taken me through PhD, post-doctoral, and senior scientist roles, including roles within flow cytometry core facilities across the UK, Ireland and Australia. That journey has shaped a deep appreciation for the technologies that enable great science.

Here at Millennium Science, our mission is to ignite life science discovery with revolutionary technology!

This blog series – which I’m calling Joe Blogs – is my way of sharing insights, practical tips, and a few behind-the-scenes stories from the technologies I work with every day. From immunoassays and cell-based analysis to real-time monitoring and flow cytometry, I’ll be unpacking the platforms and reagents that drive discovery across Australia and New Zealand’s life science community.

And here’s the best part: Joe Blogs will land every fortnight on Monday morning – your dose of lab-friendly advice to start the research week right!

What You Can Expect from Joe Blogs ✍️

Whether you’re running your first experiment on a new platform or refining an advanced workflow, Joe Blogs is designed to help you:
✅ Understand what your instruments can (and can’t) do
✅ Compare assay options with clarity and context
✅ Get practical insights from someone who’s been there
✅ Stay on top of emerging techniques and technologies

Think of it as a conversation, where we talk shop about the platforms you’re using, the questions you’re asking, and the science you’re working to advance.

Joe Blogs – Upcoming Topics 📅

Here’s a taste of what’s in the pipeline:

  • Spot the Difference: ELISpot vs FluoroSpot vs ELISA
  • Real-Time Results: What xCELLigence Really Tells You About Cell Health
  • NovoCyte or NovoCyte Opteon? Choosing the Right Flow Cytometer
  • Setting Up Your First ELISpot Assay: Tips for Success from Day One

I’d Love to Hear from You 💬

Got a burning question, a technical challenge, or a topic you’d love to see unpacked in a future post?
Let me know! I’d really like this blog to reflect the real conversations and challenges happening across our local research community.

Whether it’s about assay design, data interpretation, instrument comparisons or workflow optimisation, your ideas will help shape future editions of Joe Blogs. You can reach me directly via email or through the team at Millennium Science.

jroberts@mscience.com.au
customerservice@mscience.com.au

Let’s Keep It Real (and Local) 🤝

As part of Millennium Science, we’re proud to partner with leading life science suppliers to bring world-class tools to Australian and New Zealand researchers, with the local support to match. My goal with Joe Blogs is to give you a bit of that support upfront: advice, clarity, and maybe even a shortcut or two.

So, whether you’re measuring T cell responses, analysing cell proliferation, or exploring new approaches to data-rich experiments, you’re in the right place.

Until next time…happy experimenting!

Joe Blogs