A study led by Emma Lundberg* presents an extensive work  on the intra cellular localization of thousands of human proteins. The team generated more than 300,000 images to systematically resolve the spatial distribution of human proteins in cultivated cell lines.

Here is the abstract of this article, published in Science:

Resolving the spatial distribution of the human proteome at a subcellular level greatly increases our understanding of human biology and disease. Here, we present a comprehensive image-based map of the subcellular protein distribution, the Cell Atlas, built by integrating transcriptomics and antibody-based immunofluorescence microscopy with validation by mass spectrometry. Mapping the in situ localization of 12,003 human proteins at a single-cell level to 30 subcellular structures enabled the definition of 13 major organelle proteomes. Exploration of the proteomes reveals single-cell variations of abundance or spatial distribution, and localization of approximately half of the proteins to multiple compartments. This subcellular map can be used to refine existing protein-protein interaction networks and provides an important resource to deconvolute the highly complex architecture of the human cell.

The Cell Atlas is an open access resource: everybody can explore it !

*Emma Lundberg is associate professor at KTH Royal Institute of Technology and responsible for the High Content Microscopy facility at the Science for Life Laboratory (SciLifeLab) in Stockholm, Sweden.

As shown by AstraZeneca in nature reviews*, one third of safety failures along the drug discovery process is linked to CNS toxicity uncovered in clinical trials. To avoid this attrition, the potential neurotoxicity of any drug going through the blood brain barrier (BBB) needs to be assessed in the very early stages of new chemical entities (NCE) research. Neurotoxicity assays can be performed on the SH-SY5Y human cell line by using High-Content Screening (HCS) technologies. The present study was performed using classical 2D and 3D culture protocols. In this poster, 2D results and preliminary 3D culture results on multiple reference compounds are depicted.

View and download on Slideshare (low quality) : https://www.slideshare.net/hcspharma/in-vitro-dermocosmetology-high-content-analysis-approach-using-human-primary-keratinocytes-and-fibroblasts

Ask for high quality link  by putting your email below

3D cell models are now attracting a huge interest from scientists working both on toxicity and pharmacology assay development. They are considered more relevant at mimicking the in vivo situation. However, phenotypic assays on these models can be challenging, and are at least more complex.

A team from Molecular Devices and Cellular Dynamics has worked on the “phenotypic characterization of toxic compound effects on liver spheroids derived from iPSC using confocal imaging an three-dimensional image analyis”. The results have been published September 2016 in Assay and Drug Development Technologies, and describe how the ImageXpress Micro Confocal High-Content Imaging System and MetaXpress High-Content Image and Analysis Software (Molecular Devices) were used to manage the phenotypic characterization.

Assuming that the approach may be extensible to more complex 3D systems, such as cultures containing multiple cell types (e.g., Kuppfer cells, fibroblasts, endothelial cells), they conclude 3D analysis would allow characterization of different cell populations and their roles in toxicity and liver injury.

Source : Phenotypic Characterization of Toxic Compound Effects on Liver Spheroids Derived from iPSC Using Confocal Imaging and Three-Dimensional Image Analysis

Despite continuous improvement of DNA FISH, a method that has been extensively used for years, it still requires harsh treatments to allow probe hybridization. Oligoprobes are also expansive, and the method is time consuming. Teams are thus looking for improvements. For this purpose, Deng et al. from the Transcription Imaging Consortium (Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147) have used the CRISPR/Cas9 system to allow highly specific and efficient labeling of DNA without global DNA denaturation, which is generated by heat or chemical treatments in DNA FISH protocols.

Let’s quote the significance they attach to their work:

We have derived a new technology for the detection of genes within undisturbed nuclei of fixed cells and tissues. Previous approaches have used fluorescent DNA probes to hybridize to genes of interest, requiring treatment of heat and disruptive chemicals that distort the natural organization of the nucleus. Instead, we have used a bacterial protein, CRISPR (clustered regularly interspaced short palindromic repeats), combined with an RNA sequence as probes to find the genes of interest in the intact genome. This approach preserves the spatial relationships of the genetic elements, which are important for understanding gene expression, and the process is remarkably rapid (15 min), convenient, and can be used directly on tissues for diagnosis of disease.

This article is available here, and an explained abstract can be read on the very good “greenfluorescentblog” !

We are very pleased to welcome Kathleen to HCS Pharma. With her Master’s degree in Cellular and Molecular Biology, done at Rennes University, Kathleen was hired as an engineer at the Institut National de la Santé et de la Recherche Médicale. During those 3 years, she worked on different projects in the field of toxicology and drug screening, which gave her a strong expertise on cellular biology.

She is now our cell culture leader, welcome onboard !

Cell biology is part of the R&D activity in dermocosmetolgy. At HCS Pharma, we are developing dermocosmetology assay using our automation platform and high content analysis software.

Wound healing is a complex process resulting in new tissue formation, and skin remodeling. We have developed an in vitro automated assay in 96 well-plate using human primary epithelial keratinocytes in order to avoid expensive used of animal models, reduce the cost of assay and increase the volume of compounds tested.

This assay, also called “The scratch assay” is relevant to evaluate the efficacy of active ingredients for irritated skins as well as for stretch marks. In dermocosmetology, this assay is commonly used to assess compounds efficiency on wound healing process.


If you want to know more on this assay, follow this link to our page on the website or contact us!


The creation of a double strand break (DSB) is accompanied by the phosphorylation of histone H2AX. The measurement of serine 139 phosphorylated histone H2AX (γH2AX) is reported to be a marker of interest to identify potential genotoxic activity.

In order to evaluate the High Content Screening for γH2AX detection, 4 non genotoxic compounds and 9 genotoxic compounds from the ECVAM list I or II were selected to to be tested on HepG2 cell line. These cells offer the advantage to have H2AX expression data in the literature. In parallel, Human primary keratinocytes were included. Indeed these cells would be relevant for investigating skin adverse effects of topical applied xenobiotics with the advantage of High content imaging as valuable tool for screening in the early discovery phase.

View and download on Slideshare (low quality) :

Ask for high quality link by putting your email below

Thomas Kirchhausen, Eric Betzig et al. publish an article in Sciences about SIM (structured illumination microscopy) and the observation of moving molecules inside cells. The incredible spatial and time resolution give the opportunity to see for exemple proteins which pass the cell membrane.

On the surface of a living cell at any given time, hundreds of tiny bubbles are popping into existence, surrounding and incorporating proteins, hormones, fats, and the occasional bacteria or virus. But until now the details of this activity were inferred – you couldn’t actually see it.

Li et. al., “Extended Resolution Structured Illumination Imaging of Endocytic and Cytoskeletal Dynamics,” Science.

Source : New Instrument Captures the Secret Lives of Cells – technologyreview.com

Aditional informations : Imaging Techniques Set a New Standard for Super-Resolution in Live Cells

Aviv Regev at the Broad Institute, in Cambridge, uses fluidic systems to separate cells and submits them to detailed genetic analysis, at the rate of thousands per day. The goal is to build a comprehensive cellular human atlas, based on gene expression profiling.

The new technology works instead by cataloguing messenger RNA molecules inside a cell. These messages are the genetic material the nucleus sends out to make proteins. Linnarsson’s method attaches a unique molecular bar code to every RNA molecule in each cell. The result is a gene expression profile, amounting to a fingerprint of a cell that reflects its molecular activity rather than what it looks like.

Researchers from Yale university presented a new technology to increase presicion of high speed (and wide field) imagery. Using both LEDs and laser light, they minimize spacial coherence and granularity. It’s a promising way to optimize cells imaging.

La technologie développée par les chercheurs de Yale combine la luminosité des lasers traditionnels avec la qualité d’image permise par les LEDs, en ayant notamment recours à un laser à cavité chaotique. Une telle combinaison permet d’émettre une lumière puissante, mais dotée d’une faible cohérence spatiale, et de réduire ainsi considérablement la granularité.

Curated from www.atelier.net

Here is a nice example of cell imaging uses for stem cells characterisation. Different culture conditions are performed on adult stem cells and cell adhesion, proliferation, survival, and cell migration are followed by cell imaging.

Curated from www.jove.com

%d bloggers like this: