Instrumentation for HCS

The first reader to be developed especially for HCS was launched in 1997. The Array ScanĀ® (Cellomics, Pittsburgh, PA, USA) was the first system which integrated microscopy, digital imaging and data processing within a single platform [82].

Today, individual instruments available commercially can be classified by their optics (inverted microscope versus confocal microscope), light sources (laser versus mercury-xenon lamps) and their capabilities of performing live cell/kinetic measurements (Table 5.2).

The ArrayScan (Cellomics, Pittsburgh, PA, USA) and DISCOVERY-1 (Molecular Devices, Sunnyvale, CA, USA) contain optics based on an inverted microscope.

Confocal microscopes, used in the INCELL Analyzer 3000 (GE Healthcare) and the Opera (Evotec, Hamburg, Germany) have the capability of delivering high-resolution images.

Another HCS platform which is not microscope-based is the Acumen Explorer (TTP Labtech). This is a laser-scanning fluorescence microplate cytometer, which uses a scanning laser and optics with fluorescence filters to direct light emissions onto four photomultiplier detectors.

All HCS platforms described allow the performance of cellular assays in dense plate formats (usually 96- or 384-well plates). The Opera and the INCELL Analyzer 3000 are assembled with an internal pipetting device, which provides the capability of performing rapid kinetic measurements, even under environmental control using the INCELL Analyzer 3000.

Table 5.2 A summary of high-content screening (HCS) reader characteristics.

ArrayScan

DISCOVERY-1

INCELL

Opera

Acumen

VTI

Analyzer 3000

explorer

Optics

Inverted

Inverted

Confocal

Confocal

Photo-

microscope/

microscope/

microscope/

microscope/

multiplier

CCD camera

CCD camera

CCD camera

CCD camera

Light source

UV, Mercury-Xenon lamp

Arc lamp

Laser

Laser

Laser

Objective

5-40x

5-40x

40x

10-60X

NA

magnifications

Optics

Inverted

Inverted

Confocal

Confocal

Photo-

microscope/

microscope/

microscope/

microscope/

multiplier

CCD camera

CCD camera

CCD camera

CCD camera

Light source

UV, Mercury-Xenon lamp

Arc lamp

Laser

Laser

Laser

Plate format

96, 384

96-1536

96, 384

96-2000

96-1536

(well)

Internal environ-

No

No

Yes

No

No

mental control

Internal

No

No

Yes

Yes

No

pipetting

NA = not applicable.

The data processing is accomplished by the systems' software, which is often developed by the suppliers to provide their customers with an integrated solution for their HCS needs. A recently launched software package, entitled Cellenger (Definiens AG, Munich, Germany), not only delivers ready-to-use assays and the ability to generate novel algorithms, but also provides a database solution for handling the large data sets obtained during HCS campaigns.

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