High-Content Screening (HCS)

Discover the power of real-time analysis

HCA technology was pioneered by Cellomics in 1996, now part of Thermo Fisher Scientific, and has evolved and transformed over the past two-and-a-half decades with the customer in mind. The HCA platforms are modular and powered by our proprietary HCS Studio quantitative analysis software, which allows for sophisticated cell-level quantification and advanced statistics to be conducted at the same time imaging occurs. This real-time analysis approach, assures immediate data availability post scan, and helps researchers make informed decisions more quickly. HCS Studio was designed for biologists, not computer scientists, enabling scientists a turnkey technology to advance their imaging cytometry research. More than 2,000 peer-reviewed publications on over 30 validated assays support our HCA technology as the industry leader. Partner with us, to take advantage of the expertise and reliability of our technology, as well as decades of experience in labeling and detection with Invitrogen Molecular Probes fluorescence reagents.

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Microscopic image of cardiomyocytes in culture showing green-fluorescent tubulin intracellular structure.


Cardiomyocytes are receiving increased attention for the health safety assessment of novel drugs on the heart. Cardiovascular diseases are the leading cause of morbidity and mortality worldwide. Pluripotent stem cells are used for primary screening for potential toxicological properties and can provide an unlimited source of cardiomyocytes and therefore play a key role in cardiac regeneration. High-content analysis combines fluorescence microscopy with an automated digital imaging system to provide qualitative and quantitative analysis of changes in cellular phenotype.

Experimental details for the image above: cardiomyocytes were differentiated from IPSCs (Cat. No. A18945), plated on vitronectin- (Cat. No. A14700) coated plates, then fixed, permeabilized, and labeled with mouse anti-alpha tubulin primary antibody (green fluorescence, Cat No. A11126).

Microscopic image of hippocampal primary neurons with blue and green fluorescence
Microscopic image of neurites showing blue and green fluorescence

Neurite outgrowth

Of all the methods used to evaluate the effects of chemicals on neurodevelopment in vitro, measurement of neurite outgrowth has probably received the most attention. This is not surprising since axonal and dendritic processes are a defining characteristic of neuronal morphology and critical determinants of neuronal connectivity. Traditionally, neurite outgrowth studies relied on slow and tedious manual tracing of neurites in vitro. High-content analysis offers a much more efficient method to evaluate neurite outgrowth during compound screening studies.

Experimental details for the image above: nuclei are identified in pre- and postsynaptic vesicles within hippocampal primary neurons via Hoechst 33342 (blue, Cat. No. H3570) staining; neurites are labeled with Invitrogen Alexa Fluor 488 dye conjugated secondary antibody (green, Cat. No. A20181). Neurons were auto-identified using CellInsight CX7 LZR (autofocus, 40x objective, confocal mode).

Microscopic image of a spheroid generated from A549 cells, showing green fluorescence

3D cell imaging and spheroid assays

Cancer cells propagated in three-dimensional (3D) culture into spheroids provide a more comprehensive and extrapolative model in cancer research. Acquiring the dimensions from a larger structure includes obtaining images from different depths within the body of the spheroid and examining them in 3D or condensing into a 2D stack. Confocal imaging technology found in the CellInsight CX7 LZR is utilized in order to visualize the interior of the 3D object.

Learn more about spheroids and 3D cell culture

Experimental details for the image above: Adenocarcinomic human alveolar basal epithelial cells (A549 cells) were plated at a density of 5,000 cells/well on a Nunclon Sphera U-bottom plate (Cat. No. 174925). Spheroids were stained for 1 hour with Invitrogen Image-iT Green Hypoxia Reagent (5 μM final concentration, green, Cat. No. I14833) and Hoechst 33342 (blue, Cat. No. H3570), The image was acquired using CellInsight CX7 LZR and a 10x objective.

Fluorescence microscopy image of neurons, with red and green fluorescence indicating cell structures

Stem cell research

Stem cells are unspecialized cells that act as precursors to specific cells that are able to self-renew. They are unspecialized because they do not perform a tissue-specific function, such as carrying oxygen in the bloodstream or sending and receiving signals in the brain. Instead, they are important for generating specialized cells and helping to repair tissue. Stem cells continue to offer significant potential in regenerative medicine research efforts through the differentiation of stem cells into the desired tissue of interest.

Experimental details for the image above: neurons were differentiated from IPSCs (Cat. No. A18945), plated on vitronectin- (Cat. No. A14700) coated plates, then fixed, permeabilized, and labeled with mouse anti-alpha tubulin primary antibody (green, Cat No. A11126) and Alexa Fluor 594 dye-labeled phalloidin (red, Cat. No. A12381). Image was obtained on a CellInsight CX7 LZR instrument using a 40x objective

High-content analysis reagents

BPAE cells imaged using the CellInsight CX5

BPAE cells imaged using the Thermo Scientific CellInsight CX5 platform and stained with DAPI (blue), Invitrogen Alexa Fluor 568 phalloidin (red), goat anti–mouse IgG (H+L) secondary antibody, and Alexa Fluor 488 conjugate (green).

A549 cells imaged using the CellInsight CX7 platform

A549 cell image acquired using the CellInsight CX7 platform and stained with Invitrogen HCS CellMask Blue (blue), Alexa Fluor 488 Phalloidin (green), and Alexa Fluor 750 goat anti–mouse IgG secondary antibody (pink).

HT1080 cell image acquired using an ArrayScan reader

HT1080 cell image acquired using an ArrayScan system and stained with DAPI (blue), Invitrogen Alexa Fluor 555 phalloidin (red), and anti-tubulin and Alexa Fluor 647 goat anti–mouse IgG antibodies (green).

HCS reagent selection guides

Drawing on decades of experience in fluorescence imaging, Invitrogen HCS products are developed using Thermo Scientific high-content platforms with special considerations for the high-throughput workflow and automated imaging:

  • Cell and nuclear masks for automated demarcation
  • Robust functional probes for cell health interrogation
  • Flexible assay workflow for automated processing
  • Broad fluorophore choice for easy multiplexing
  • Validated on multiple imaging platforms

Select from the reagents optimized for use with your HCS platform:

Other cell structure and segmentation tools

Clear identification of cell and nuclear boundaries in automated imaging requires fiducial markers that will label cellular structures robustly and reproducibly. Cell masks are designed specifically for automated imaging. Invitrogen Molecular Probes cell structure labels are widely used in microscope imaging and HCS.

See reagents for cell structure

Other cell health and cytotoxicity assays

We offer a diverse selection of imaging and microscopy assays for the analysis of cell viability, proliferation apoptosis, cell cycle and DNA repair, migration, adhesion and other cellular functions. Many of these assays have been validated specifically for HCS platforms.

Browse cell health and cytotoxicity assays by application: