High Content Screening (HCS)

 

With any form of visual study it is possible that apophenia (patternicity) can result, where the observer collects data and unknowingly fits an erroneous connection or theory. We have evolved the ability to discern patterns in visual data often by viewing a small portion of a data set, in the laboratory this can be described as unwittingly looking for the front cover of Nature, where the researcher is looking for cells that demonstrate the ideal rather than collecting representative data. Hence when observing a cellular response to stimuli, time must be taken to screen a whole population in an unbiased fashion, to understand variation and the population norm, consequently automation is required.

High Content Screening (HCS) allows for the setting of imaging and analysis criteria then allowing automation in batch format to visualise and numerate across multiple analysis parameters. In the laboratory around 30-70 multi-well plates are processed on a weekly basis via automated confocal screening and flow cytometry (traditional or imaging) 


 

PerkinElmer Opera Phenix, a modified spinning disk confocal High Content Screening system with a dedicated sCMOS camera for each imaging channel so to allow rapid data throughput. The system allows  a range of magnifications and resolutions up to a x60 objective lens with water immersion. There is the possibility of stacking up to 42 multi-well plates at room temperature for high throughput of fixed cell samples and a 42 multi-well plate stacker in an automated incubator. The system is well suited for 3D culture, spheroids, organoid development as well as more traditional 2D methods. 

 

The Operetta CLS has a sensitive sCMOS camera and 8 high powered light-emitting diodes (LEDs) to give optimised excitation and emission spectra’s of a wider range of wavelengths  with the aim of brightening up 2D, 3D, 4D HCS.Automated analysis using PerkinElmer Harmony and Columbus platforms gives high through-put analysis, vital for the fast analysis of thousands acquired images.  This gives a vast array of read-outs including intensity, morphology and texture variations, allowing large cell populations to be comprehensively investigated.




Using the Columbus software image processing and analysis of many thousands of images can be completed with quickly and in a standardised manner. The image above shows  (Fig.1.) C918 cells stained with the nuclear stain Hoechst and the mitochondria dye MitoTracker Green,  Fig.2. the detection of each individual nuclei. Fig.3. has highlighted the detection of the mitochondria in the cytoplasm whilst Fig.4  shows the detection of a subpopulation of cells in green and any excluded cells in red.  
 


 

Institute equipment

The institute has been analyzing data from multi-well plates in a high throughput/content format for over ten years in the fields of drug discovery and biomarker discovery via automated microscopes or complete systems. There an an array of systems for microscopy, flow cytometry and histology screening

  • PerkinElmer Opera Phenix with multi-well plate loading automation (room rg106)
  • PerkinElmer Operetta CLS, a fluorescence wide-field based system for screening system multi-well plates. (MCRC 1st floor mid.lab).
  • ThermoScientific Insight, wide field system with automated plate loader (room rg106)
  • ImageStream, imaging cytometry
  • BD Canto with high throughput carousel
  • BD Fortessa FACS analysis with High Throughput/High Content multi-well plate loader
  • PerkinElmer Vectra, an automated histology based system for imaging up to seven fluorescent labels per tissue section


Software

  • PerkinElmer Harmony (data analysis)
  • PerkinElmer Columbus (data management and analysis)
  • SpotFire HCS (multi-plate assay analysis)
  • PerkinElmer Inform and Phenochart
  • Definiens Tissue Studio and Image Miner

The facility offers assistance in the visualisation, collection of data, and the analysis of the population whereas the Scientific Computing facility offers a fully data storage, backup and archival service.