HCS

High Content Screening (HCS) is multifactorial quantitative analysis method for cellular samples, based on automated microscopic imaging acquisitions.

The bio-applications used for the analyses make it possible to obtain simultaneously a large number of multiparametric quantitative data related to targeted cellular phenotypes (morphology, presence and/or intensity of marking, intra-cellular location,…). The automation of microscopic acquisitions allows the simultaneous screening and analysis of many molecules, conditions or cell lines from multi-well plates.

Combined with the use of robotic arms for loading the plates into the machine, this technique becomes an extremely powerful analytical tool that complements High Throughput Screening strategies for screening pharmaceutical molecules.

To this end, the High Content Imaging cluster of the ISTEM Cellular Imaging Platform brings together several imagers, each with different potentialities and approaches.

 

  • Cellinsight CX7 / Arrayscan VTi (Thermofisher Scientific)

The arrayscan VTi and the CX7 are two versions of the same machine, which run in parallel on the platform, with the same software suite (HCS Studio). Two additional analysis licenses are also available on the platform, for consultation and reanalysis of results if necessary.

 

As technology has evolved between these two machines,some differences can be noted in the optical and mechanical characteristics of the two machines:

Although it is not possible to create new ones, about twenty bio-applications are available, and allow to set up analyses of the types Neuritic Analysis, Cell Cycle, Collocation, aggregates detection, etc….

The CX7 is associated with an Orbitor robotic arm that allows the automated loading of a dozen plates into the machine, making this machine compatible with screening studies, and work outside working hours.

 

CX7 Arrayscan
Illumination LED HBO (Mercury Lamp)
Filters 305,350,488,555,590,647,750 DAPI,GFP,DSRED,TexasRed, Cy5
Magnifications X10; x20; x20 (NA 0.45); X40 X5; x10; x20; x40; x63
Methods Epifluorescence, Brightfield, Confocal Epifluorescence

 

 

  • ImageXpress (Molecular Devices)

 

This HCS Imager is distinguished from the CX7/Arrayscan by the ability to work with living cells thanks to a thermostat-controlled enclosure and in a controlled atmosphere at 5% CO2. It also has an automated pipetting head for dispensing drugs in a well, for example.

The MetaXpress software, based on Metamorph, also allows the use of analysis modules similar to pre-programmed Bioapplications, but it is also possible to create all or part of a bio-application using the Custom Module Editor.

the optical characteristics of the machine are as follows

 

 

ImageXpress
Illumination LED
Filters 305,350,488,555,590,647,750
Magnifications X10; x20; x20 (NA 0.45); X40
Methods Epifluorescence

 

Two analysis stations complete the acquisition/analysis station attached to the machine. As with the CX7, the imageXpress is equipped with a robotic arm for the automated handling of multiple multi-well plates.

 

  • Incucyte Zoom and Incucyte S3 (Sartorius)

 

 

The Zoom and the S3 are two versions of the same imager from Sartorius.

These Incucytes, installed directly in culture incubators, allow to follow and acquire images of living cells in situ, in fluorescence (green or red) or in phase contrast, over short periods or for several weeks, in an optimal environment.

Each of these machines can accommodate up to 6 multi-well culture plates and the image acquisition frequency can be set by the user.

 

 

 

The photos can be viewed and exported from the central unit attached to each incucyte. Depending on the machine under consideration, different analysis modules are availables for quantitative analysis:

  • morphology and confluence of culture
  • Scratch Wound (Scratch Wound)
  • of Chemotaxis
  • of neuritic analysis
  • of spheroid size