Common equipment in HTS labs

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Common Equipment in HTS Labs
From The NIH Chemical Genomics Center Assay Guidance Manual

1 Microplates and Microplate Readers
2 Liquid Handling Devices

Microplates and Microplate Readers

Most commonly used microplates have 96-wells (12 columns x 8 rows) with 250-300 volume µL capacity. Recent introductions include 96-well half area, 384- and 1536-well plates using the same outside plate dimensions (or foot print). The 96-well half area and 384-well plates use less reagents (75-100 µL capacity), and a major advantage is the reduced reagent consumption. However, these plates require special care during reagent delivery. The 1536-well plates are used less commonly, and require special liquid delivery equipment.

Microplate Types:

Clear bottom
Black Opaque
White Opaque
Mixed formats
CytoStar-T
FlashPlates
Scinti-Strips

Plate Configurations:

24-well
96-well
96-well half-area
96-well High efficiency (HE) plates
384-well
1536-well

CytoFluor 4000 Fluorescence Multi-well Plate Readers

Assays for quantifying cell-associated or soluble fluorescence can be measured in a CytoFluor 4000 Fluorescence Multi-well Plate Reader. These systems have the sensitivity and flexibility for low level detection, which is a must for fluorescent assays. CytoFluor Plate Readers combine detection of less than 3 pg/well of fluorescein with a five-log dynamic range. You can adjust the gain settings to match the fluorescent signal of each assay.

Even lower detection limits are possible with enzyme amplified assays - as few as several hundred attomoles of analyte can be quantified. Almost any fluorescence assay can be automated on the CytoFluor Plate Reader, which comes with 4 standard excitation and 5 standard emission filters. For specialized applications, you can choose from 15 additional optical filters. Standard multi-well plates and strips of all shapes and popular well formats can be read. Custom-designed plates and Terasaki plates can also be read using special adaptors. You can even choose to read the plates from the top or the bottom. Changing the direction of the read is easy, and the same plate can be scanned up to 200 times at user-defined intervals.

To help you with small volume assays or long term assays, a plate can be shaken automatically within the instrument - for precisely the amount of time you specify. Optional design features keep the CytoFluor Plate Reader in step with the latest applications:

Temperature Control
A temperature control option is available to run temperature-dependent kinetics.
High-throughput Screening (HTS).
Reads up to 6000 samples per hour per instrument, so you can run your screening assays on the same instrument on which they were developed.
Robotics Friendly.
Easily integrates with laboratory robotics and LIMS for greater productivity.
Special Plate Reading Capability.
Reads Terasaki plates for small volume (10 - 20 µl/well) applications.

Fluorescent - Imaging Plate Reader (FLIPR)

Two 96-well FLIPRs and a 384-well FLIPR instruemnts are available at Sphinx. Neuroscience Dircovery Research has one 96-well plate reader in Indianapolis. Utilization of functional, high-throughput assays on living cells allows a combination of screening and pharmacological information to be obtained in a single assay, improving and speeding the generation of key discovery data. FLIPR® (Fluorometric Imaging Plate Reader) technology is an industry leader in using live cell assays in screening operations.

Now you can get the sensitivity, data fidelity and real-time, kinetic information of the FLIPR system in a 384-well mode complete with 384 simultaneous fluid additions. With enhanced automation features allowing it to be part of a linear robotics system, the FLIPR384® system stands at the forefront of today's rapid-throughput screening

technologies.

The ability to monitor. Real-time, kinetic data also provides additional pharmacological information for ranking relative potencies of drugs and gives information on the kinetics of the drug-receptor interaction.

General system specifications:

Simultaneous optical measurements on all wells of a 384- or 96-well plate at rates of up to once per second.
Interchangeable pipettor head for simultaneous dispensing to either 384 or 96 well plates.
Semi-automated disposable tip loading for both 384 and 96-well pipettor heads.
Integrated tip washing station to decrease number of tip changes, saving time and money.
Six pipettor-accessible positions on the FLIPR384® system platform.
Computer controlled front panel for access to plate and tip stations.
Water cooled, argon-ion laser illuminated patented FLIPR/FLIPR384® system optical detection scheme.
Cooled CCD detection optics.
Windows™ NT-based FLIPR Control software.
Three primary configurations (with options):
- Maual
- Heated stage-Robot line
- Heated stage-Stacker fed

Key fluorescence intensity assays:

Intracellular calcium
Membrane potential
Intracellular pH
Intracellular sodium

Key application areas:

Primary/secondary screening
Assay development
Structure-activity relationships

LJL Analyst™

Analyst™ AD Assay Development System is a flexible system for the development of assays destined for High Throughput Screening (HTS). Analyst AD gives assay development groups a time- and cost-effective way to speed the transfer of assays to HTS. By sharing a common detection hardware and software system with Analyst™ HT (for HTS using the 96- and 384-well format), Analyst AD eliminates the need to modify and revalidate methods due to differences in detection system features and performance.

Analyst AD supports assay detection using:

Fluorescence Intensity
High Efficiency Fluorescence Polarization (HEFP™)
Time-Resolved Fluorescence
Luminescence

Assay detection methods control all instrument parameters effecting superior performance in any single mode. However, the user is not limited to assays that use one wavelength or even one mode. Analyst AD also supports two-wavelength assays (such as TR-FRET) or two method assays.

For each detection modality, sensitivity, dynamic range, signal-to-noise and cross-talk are all optimized. Analyst AD delivers uncompromising sensitivity and dynamic range in all

fluorescence-based assay detection modes. Analyst AD achieves this performance with SmartOptics™, a unique system which selects and configures light sources, filters, detectors, and optical paths to insure maximum assay performance. Using novel three-axis positioning, light is focused and collected in a small "sensed volume" for maximum performance, excluding spurious light that contributes to noise and cross-talk.

In the case of standard 96-well microplates, the sensed volume is a small fraction of the total assay well volume. The sensed volume remains the same for 384-well microplates, ensuring essentially identical assay performance in 96- and 384-well formats. Ultimate performance in low volume assays is achieved by using Analyst AD with LJL's HE™ microplates. The SmartOptics system and HE plates have been designed together to deliver the same high sensitivity and low background in a 10 microliter assay in an HE plate compared to a 200 microliter assay in a traditional 96-well microplate.

Wallac VICTOR² Multilabel Plate Reader

The Wallac 1420 VICTOR² is a multilabel, multitask plate counter designed to support the demands of academic and industrial laboratories for assays based on all types of label. An extended version of the successful Wallac VICTOR multilabel counter, the new VICTOR² allows immediate access to more than 10 counting modes, covering all of the main non-radioactive counting technologies. VICTOR² accepts all types of microtitration plates with between 1 and 864 wells, as well as Petri dishes, slides, filters and Terasaki plates. All models include scanning, shaking and kinetics modules. To support individual applications in cell or molecular biology, binding studies, environmental and food testing, toxicology and drug screening, VICTOR² can also be supplied with dispenser module, temperature control, bottom reading, high density reading and various other options.

A small bench top unit, VICTOR² is easy to place as a stand-alone instrument or

to integrate into a robotic system. Performing the work of a fluorometer, a luminometer, a time-resolved fluorometer and a photometer, it provides a superior alternative to a whole series of mutually incompatible analytical instruments. Ease of use and general reliability are enhanced by 32-bit Windows Workstation software, designed to utilize the full capabilities of Windows NT and Windows 95.

The Wallac 1420 VICTOR™ F fluorometer is an advanced detector for Fluorescence based assays on microplates. High quality VICTOR™ optics provide reliable measurement of a wide variety of assay types.

VICTOR™ F accepts all types of microtitration plates with between 6 and 1536 wells. Filterplates and membranes, Petri dishes, slides and Terasaki plates are all measured. Non-standard configurations can also be defined using the instrument's 32-bit Windows Workstation software. The most versatile counting solution for all types of cell-based assay, VICTOR™F offers specialties such as scanning function, top and bottom measurement, shaking in three different modes and dual ratio measurements. Similar, thoughtful features add distinction to your work within the whole gamut of molecular biology applications, as well as in binding studies, environmental testing, and especially useful in kinetic assay, VICTOR™ F allows delays and repeats by plate, row or well.

TopCount®

The TopCount® is a microplate reader that detects luminescence signals (other than fluorescence.). The HTS version lets you count radioisotopic and luminescent samples twelve at a time in both 96- and 384-well microplates, separately or intermixed. It allows for counting and analyzing assays in the 96-well format while you convert them to the 384-well microplate, without losing productivity. The external plate stacker allows more than 15,000 samples to be loaded for the highest unattended counting capacity, or you can integrate your TopCount NXT into a robotic system for high throughput applications. Being Windows NT-based, TopCount NXT is easily networked to provide error-free, automatic data transmission. The Hologram database automatically stores the data for every sample counted, and avoids loss of data due to network and printer failures.

Cross-sectional diagram of the TopCount counting a typical opaque microplate; illustrating the elimination of crosstalk. TopCount NXT analyzes two, six, or twelve samples at a time, and counts samples in 24- and 96-well microplates, as well as microcentrifuge tubes. Only TopCount NXT can control the counting chamber temperature to ensure a reproducible counting environment for all your assays.

Applications: count gammas, betas, alphas, or luminescent samples, TopCount NXT will

do it without interfering crosstalk. Perform solid scintillation, liquid scintillation, filter counting, Cerenkov counting, SPA®, and scintillatingmicroplate counting, without compromising quality. And, TopCount NXT’s high efficiency count mode significantly increases sensitivity for a variety of assays.

MicroBeta TriLux® and MicroBeta JET®

MicroBeta TriLux® is a compact, complete automatic multidetector Liquid Scintillation Counter (LSC).It counts directly from microtitration plates as well as from filters and tubes. By combining LSC reliability with plate reader simplicity, it allows vast savings in time, consumables and waste. MicroBeta TriLux is ideal for samples labelled with 3H, 14C, 32P, 33P, 35S, 51Cr, 125I, and glow- type luminescence labels as well as for scintillation proximity assay. Also available, a new version of MicroBeta has built-in injectors to allow also the use of flash-type luminescence reagents.

MicroBeta JET is the world's only multi-detector luminescence counter with reagent injectors for using flash-type luminescence reagents. Like MicroBeta TriLux, JET can also be used for beta and gamma labels. Also like TriLux, JET can measure from microtitration plates, tubes or filters in most formats. MicroBeta JET is available in 6, 3, 2 or 1 detector models. The single detector model can have up to four injectors and multi-detector models up to two injectors per detector. There are 16 or 32 plate versions of JET, as well as a robot loading model.

Liquid Handling Devices

Liquid handling equipment includes single and multi-channel pipetors used manually, and programmable fluid delivery devices. These devices can precisely and repeatedly dispense a few to several hundred microliters of aqueous solutions when properly calibrated. These devices employ disposable or polyethylene pipet tips, and are not suitable for dispensing highly corrosive or viscous liquids accurately. Examples of such liquids include concentrated acids, bases, some organic solvents, oils, >40% glycerol, undiluted blood, serum or plasma and other viscous material.

Hand-held pipets:

Single-channel
Multi-channel
Electronic (i.e. Matrix)
Repeaters

Automated pipeting devices:

Multi-drop
Biomek 1000/2000
Multimek
MRD8/MRD96
PlateMate
Work Stations
Integrated Robotic systems
Nanoliter dispensers
Units with plate stackers
Tecan

The quality of assay data depend critically on the ability of individual scientists to use the liquid delivery devices appropriately. It is highly recommended that you should make yourself familiar with all pipeting equipment in your lab. Also note that these liquid dispensors will have to be calibrated on a regular basis. For automated liquid handlers, special procedures recommended by the manufacturers are employed. These procedures usually involve serial dilutions of dyes from stock solutions and the determination of the accuracy and precision of the dilutions.

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