High-Throughput Screening for Flow Cytometry


High-Throughput Screening

High-throughput screening (HTS) is an essential part of the current drug discovery process. It allows for rapid automated testing of large libraries of biological or chemical compounds to assess their effect on a specific biological target. Once a lead molecule has been identified, additional testing is required to further characterize lead compounds and assess their potential efficacy.

Antibody Screening

high-throughput screening imageCreating a highly specific and novel biologic involves high-throughput antibody screening of hundreds of candidate clones. An unparalleled combination of speed, sensitivity, and multiparameter analysis are all required when assessing antibody clones to find those that bind strongly and specifically to target antigens with the potential to become commercialized products.

Phenotypic Screening

Phenotypic screening is often described as a target agnostic approach to drug discovery. Here, molecules are assessed for their ability to promote a desired phenotype associated with a therapeutic effect. Typically, phenotypic screening is used in small molecule drug discovery often utilizing cellular models of human disease pathophysiology. It may encompass multiple parameters to assess the likely efficacy of drug candidates including elucidation of a desired effect as well as monitoring of adverse events such as toxicity.

Lead Optimization

Once a lead molecule has been selected, it is necessary to evaluate its characteristics and performance. This can involve several preclinical assays including safety, pharmacokinetics (PK), Absorption, Distribution, Metabolism, Excretion (ADME), internalization, and functional studies. Functional assays may include neutralization, agonism, antagonism, proliferation, and apoptosis assays.

Flow Cytometry as a Tool for Screening

To perform this array of tests throughout the whole drug discovery process, flexible and sensitive instrumentation is needed to manage the required assay speed, without sacrificing high-parameter data collection and rare-event detection. For these reasons, flow cytometry has become the gold standard for performing antibody and phenotypic screening as well as characterization assays. Importantly, flow cytometry allows for unparalleled multiplexing, while providing the flexibility, speed, and reproducibility to analyze a high number of individual events and determine their distributions.


Challenge of High-Throughput Antibody Screening & Analysis

In the drug discovery process, it is necessary to screen for candidate hits, filter those hits into lead candidates, and then analyze each candidate’s clinical potential. Flow cytometry is an excellent technique to perform the necessary experimental volumes and conditions. There are three important characteristics of high-throughput flow cytometry workflows: multiparameter/multiplexed analysis, acquisition speed, and automation readiness.

Automated HTS

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Combining High-Parameter, High-Speed Flow Cytometry with Automation

Collecting data on multiple parameters at once enables more complex and informative multiplexed analysis. This can be achieved by using flow cytometers with multiple lasers and detectors. Fundamentally, the more lasers and parameters are involved the more data is collected each time.

Acquisition speed describes the rate at which a flow cytometer moves, counts, and collects data. Speed of acquisition is a key variable for improving the detection of rare events, such as T-cell and other immune-cell population changes. By collecting more data within a limited amount of time, the likelihood of capturing these rare events increases. For optimal results, select a flow cytometer that avoids sample carryover by automatically washing the probe between samples.

Flow cytometry instruments built with automation in mind can also help actualize the high-throughput potential of an instrument, as analysis demands increase further. Flow cytometers ready for robotic integration and with automation features can simplify operations, save time, and improve lab performance.

High-Throughput Screening Instruments

Introducing the Market Leading Instrument for HTS

We know you are under increasing pressure to deliver ever more data, more quickly and that you need the right tools to support you. In the past, you may have had to compromise between speed, the number of parameters measured, the number of events collected, or even data resolution.

With the ZE5 Cell Analyzer, the market leading HTS instrument, this is no longer the case. Achieve unprecedented speed without having to compromise on performance. You can screen your samples faster and with better data quality than with any other flow cytometer.

A Screening Flow Cytometer without Limits

  • High-throughput ready hardware and software
  • Maximal productivity with integrated automation and 24/7 operation
  • High-speed acquisition without compromising data quality
  • Superior assay design and instrumentation flexibility

The impressive features of the ZE5 Cell Analyzer enable you to design experiments that no other flow cytometer available today can do. Five lasers and 30 detectors, fast sampling from 96-well or 384-well plates, and speed up to 100,000 events per second means you can design larger panels and run them at unparalleled speeds.


Speed up Your Discovery without Compromise

  • automation robotic arm icon


    Integrates with laboratory robotics system via a device-agnostic application programming interface (API)

  • run icon

    96-well plates in 15 min
    384-well plates in 50 min

    ensuring low carryover at high speed with high-throughput screening

  • speed icon

    100,000 EVENTS per sec

    without data loss or compromised resolution

  • 24/7 turning cog icon

    24/7 with a single operator

    using high-capacity fluidics tanks


Built for Screening

Efficient and Flexible Sample Handling

Plate inside ZE5 Cell Analyzer

For high-throughput a plate loader is essential - the ZE5 Cell Analyzer enables:

  • Hands-free hardware switch over — switch between plates formats or tubes using the integrated sample loader
  • Compatibility with any plate format — round bottom, flat bottom, V bottom, deep-well
  • Sample preservation — on‑board temperature control and orbital agitator
  • Removing Obstacles

    • Prevent blockages — with the innovative bi-directional sample pump and superior fluidics
    • Minimize carry-over — the sample line is cleaned between each sample quickly and automatically
    • Elimiate manual well parsing — sample parsing is done on the fly as the data is collected
    Carryover reduction - Figure 1 image

    Figure 1. Carryover reduction.
    , lysed whole blood was run on the ZE5 Cell Analyzer in high-throughput (HT) mode. After each sample, the system performed and automatic wash cycle of 0.25 sec outside and 1.75 sec inside the sample injection port (SIP). A clean tube of water was run immediately after the wash to evaluate carryover. B, the resulting carryover data shown average carryover of 0.046% (+ 0.023%). Carryover = (Count-wash well- / Count - Sample well - ) x100wash well /Sample well100

24/7 Operation with Large-Capacity Fluidics Upgrades

 Large-Capacity Fluidics

Go for longer without filling back up

  • 20-liter level-sensed external fluidics cart
  • Direct house dH2O connection as sheath source

Seamless Integration into Robotic Workcells and Laboratory Information Management Systems (LIMS)

ZE5 Cell Analyzer combined with robotics

Unleash the full potential of the ZE5 Cell Analyzer by combining it with robotics

  • Automate plate loading using robotics within a workcell
  • Integrate with LIMS and scheduling software through an application programming interface (API)
  • Experience a vendor-agnostic API allowing seamless integration with a wide range of automation providers for full customization
  • Contact a Specialist

    Unlock the Power of High-Throughput Screening

    Contact a HTS specialist to discuss how the ZE5 Cell Analyzer can help you harness the power of high-throughput screening with your experiments

    Contact a HTS Specialist

Antibodies & Reagents

Suitable antibodies and reagents are critical components for HTS. Bio-Rad is dedicated to providing cutting edge reagents for all flow cytometry applications, and has recently launched an exclusive range of StarBright Dyes to allow you to build superior panels. Bio-Rad also has an extensive range of high-quality antibodies and reagents validated for flow cytometry, to help you conduct successful experiments.

StarBright Dyes image

High-Throughput Screening Data

Acquisition Speed & Data Quality

Dragon Beads from Bangs Laboratories Inc were acquired on the ZE5 Cell Analyzer and other competitor instruments at serial dilutions. The ZE5 Cell Analyzer shows consistent linearity between expected and measured event count up to 100,000 events per sec (eps), whereas other systems drop off around 20,000 eps due to significant data abort.

consistent population resolution

FITC-labeled beads were acquired on the ZE5 Cell Analyzer at increasing event rates from 4,000–129,000 events per sec (eps) and showed consistent population resolution as determined by %CV.

  • Fast Plate Sampling — Competitive Comparison

    fast plate sampling - ze5 v competitor

    Samples were run on a 384-well plate with the ZE5 Cell Analyzer and a leading competitor. 10 µl sample acquisition volume, HT mode, flow rate 2 µl/sec, 5 sec agitation on column 1 and 13.

    The ZE5 Cell Analyzer finishes the 384-well plate in 50 min; the competitor system took 90 min.

Carryover Rate Analysis

data image

FITC-stained cells were run on the ZE5 Cell Analyzer in HT mode, alternating blank and cell wells in eight replicates. Carryover rate is calculated as cells-of-interest in blank/cells-of-interest in sample well. The average carryover rate is 0.007%.

carryover rate analysiscell count carryover

See more data: Download "Get more from your experiments PDF"

Our Automation Partners

The cytometer's application programming interface (API) facilitates integration into any workcell, so you can power through screening campaigns faster and with larger panels than ever before. We can advise you on your project, as we have already worked with many automation providers to integrate the ZE5 Cell Analyzer into robotic workcells

Screening Uncovered

Evolution of Flow Cytometry as a Screening Platform

Flow cytometry has evolved, the latest generation of screening flow cytometers provide easy-to-use, high-plex assays at speeds only previously seen using ELISA and imaging cytometry systems. Learn about the advantages of flow cytometry and the superior capabilities of the ZE5 Cell Analyzer.


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  • Acquisition to Analysis

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  • Contact a Specialist

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    Contact a HTS expert to discuss how the ZE5 Cell Analyzer can help you increase the efficiency of your experiments

    Contact a Specialist
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