High Content Screening
441 views | +0 today
Follow
Your new post is loading...
Your new post is loading...
Scooped by CYTOO
Scoop.it!

High-throughput, multiparameter analysis of single cells

High-throughput, multiparameter analysis of single cells | High Content Screening | Scoop.it

Heterogeneity of cell populations in various biological systems has been widely recognized, and the highly heterogeneous nature of cancer cells has been emerging with clinical relevance. Single-cell analysis using a combination of high-throughput and multiparameter approaches is capable of reflecting cell-to-cell variability, and at the same time of unraveling the complexity and interdependence of cellular processes in the individual cells of a heterogeneous population. In this review, analytical methods and microfluidic tools commonly used for high-throughput, multiparameter single-cell analysis of DNA, RNA, and proteins are discussed. Applications and limitations of currently available technologies for cancer research and diagnostics are reviewed in the light of the ultimate goal to establish clinically applicable assays.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Improving Access to HC Screening Images

Improving Access to HC Screening Images | High Content Screening | Scoop.it

High-content screening (HCS) is a powerful technology that addresses a variety of questions within biological research. Established for pharmaceutical small molecule screening, it is regularly applied in target discovery and validation. Capable of observing phenotypic changes, HCS requires a technically complex infrastructure of hardware and software components. This complexity is compounded by the management of HCS images and image-analysis results, which can be resource- and time-consumptive.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

An Open Source Based High Content Screening Method for Cell Biology Laboratories Investigating Cell Spreading and Adhesion

An Open Source Based High Content Screening Method for Cell Biology Laboratories Investigating Cell Spreading and Adhesion | High Content Screening | Scoop.it
Background

Adhesion dependent mechanisms are increasingly recognized to be important for a wide range of biological processes, diseases and therapeutics. This has led to a rising demand of pharmaceutical modulators. However, most currently available adhesion assays are time consuming and/or lack sensitivity and reproducibility or depend on specialized and expensive equipment often only available at screening facilities. Thus, rapid and economical high-content screening approaches are urgently needed.

Results

We established a fully open source high-content screening method for identifying modulators of adhesion. We successfully used this method to detect small molecules that are able to influence cell adhesion and cell spreading of Swiss-3T3 fibroblasts in general and/or specifically counteract Nogo-A-Δ20-induced inhibition of adhesion and cell spreading. The tricyclic anti-depressant clomipramine hydrochloride was shown to not only inhibit Nogo-A-Δ20-induced cell spreading inhibition in 3T3 fibroblasts but also to promote growth and counteract neurite outgrowth inhibition in highly purified primary neurons isolated from rat cerebellum.

Conclusions

We have developed and validated a high content screening approach that can be used in any ordinarily equipped cell biology laboratory employing exclusively freely available open-source software in order to find novel modulators of adhesion and cell spreading. The versatility and adjustability of the whole screening method will enable not only centers specialized in high-throughput screens but most importantly also labs not routinely employing screens in their daily work routine to investigate the effects of a wide range of different compounds or siRNAs on adhesion and adhesion-modulating molecules.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

A functional system for high-content screening of neuromuscular junctions in vitro

A functional system for high-content screening of neuromuscular junctions in vitro | High Content Screening | Scoop.it
High-content phenotypic screening systems are the logical extension of the current efficient, yet low information content, pre-clinical screens for drug discovery. A physiologically accurate in vitro neuromuscular junction (NMJ) screening system would therefore be of tremendous benefit to the study of peripheral neuropathies as well as for basic and applied neuromuscular research. To date, no fully-defined, selective assay system has been developed which would allow investigators to determine the functional output of cultured muscle fibers (myotubes) when stimulated via the NMJ in real time for both acute and chronic applications. Here we present the development of such a phenotypic screening model, along with evidence of NMJ formation and motoneuron initiated neuromuscular transmission in an automated system. Myotubes assembled on silicon cantilevers allowed for measurement of substrate deflection in response to contraction and provided the basis for monitoring the effect of controlled motoneuron stimulation on the contractile behavior. The effect was blocked by treatment with D-tubocurarine, confirming NMJ functionality in this highly multiplexed assay system.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Benchmarking of Multivariate Similarity Measures for High-Content Screening Fingerprints in Phenotypic Drug Discovery

High-content screening (HCS) is a powerful tool for drug discovery being capable of measuring cellular responses to chemical disturbance in a high-throughput manner. HCS provides an image-based readout of cellular phenotypes, including features such as shape, intensity, or texture in a highly multiplexed and quantitative manner. The corresponding feature vectors can be used to characterize phenotypes and are thus defined as HCS fingerprints. Systematic analyses of HCS fingerprints allow for objective computational comparisons of cellular responses. Such comparisons therefore facilitate the detection of different compounds with different phenotypic outcomes from high-throughput HCS campaigns. Feature selection methods and similarity measures, as a basis for phenotype identification and clustering, are critical for the quality of such computational analyses. We systematically evaluated 16 different similarity measures in combination with linear and nonlinear feature selection methods for their potential to capture biologically relevant image features. Nonlinear correlation-based similarity measures such as Kendall’s τ and Spearman’s ρ perform well in most evaluation scenarios, outperforming other frequently used metrics (such as the Euclidian distance). We also present four novel modifications of the connectivity map similarity that surpass the original version, in our experiments. This study provides a basis for generic phenotypic analysis in future HCS campaigns.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Functional Screening Assays with Neurons Generated from Pluripotent Stem Cell–Derived Neural Stem Cells

Rapid and effective drug discovery for neurodegenerative disease is currently impeded by an inability to source primary neural cells for high-throughput and phenotypic screens. This limitation can be addressed through the use of pluripotent stem cells (PSCs), which can be derived from patient-specific samples and differentiated to neural cells for use in identifying novel compounds for the treatment of neurodegenerative diseases. We have developed an efficient protocol to culture pure populations of neurons, as confirmed by gene expression analysis, in the 96-well format necessary for screens. These differentiated neurons were subjected to viability assays to illustrate their potential in future high-throughput screens. We have also shown that organelles such as nuclei and mitochondria could be live-labeled and visualized through fluorescence, suggesting that we should be able to monitor subcellular phenotypic changes. Neurons derived from a green fluorescent protein–expressing reporter line of PSCs were live-imaged to assess markers of neuronal maturation such as neurite length and co-cultured with astrocytes to demonstrate further maturation. These studies confirm that PSC-derived neurons can be used effectively in viability and functional assays and pave the way for high-throughput screens on neurons derived from patients with neurodegenerative disorders.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Assessment of the in vitro γH2AX assay by high content screening as a novel genotoxicity test

The γH2AX assay is widely used as a marker of DNA damage in multiple scientific fields such as cancer biomarker, clinical studies and radiation biology. In particular, the in vitro γH2AX assay has been suggested as a novel in vitro genotoxicity test with potential as a pre-screening tool. However, to date, limited assessments have been carried out to evaluate the sensitivity, specificity and accuracy of the in vitro γH2AX assay.

 

In this study, the microscopy-based system combining automated cellular image acquisition with software quantification for High Content Screening (HCS) has been used for the first time to evaluate the in vitro γH2AX assay. A panel of well-characterised genotoxic and non-genotoxic compounds was selected to assess the performance of the in vitro γH2AX assay in the human bronchial epithelial cell line BEAS-2B.

 

The results obtained during this preliminary assessment indicate that the in vitro γH2AX assay has a high accuracy (86%) as a result of high sensitivity and specificity (86–92% and 80–88% respectively). Our data highlight the potential for γH2AX detection in HCS as a complement to the current regulatory genotoxicity battery of in vitro assays. We therefore recommend more comprehensive assessments to confirm the performance of the in vitro γH2AX assay by HCS with a more extensive set of compounds.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

A high content screening assay to predict human drug-induced liver injury during drug discovery

A high content screening assay to predict human drug-induced liver injury during drug discovery | High Content Screening | Scoop.it

Adverse drug reactions are a major cause for failures of drug development programs, drug withdrawals and use restrictions. Early hazard identification and diligent risk avoidance strategies are therefore essential. For drug-induced liver injury (DILI), this is difficult using conventional safety testing. To reduce the risk for DILI, drug candidates with a high risk need to be identified and deselected. And, to produce drug candidates without that risk associated, risk factors need to be assessed early during drug discovery, such that lead series can be optimized on safety parameters. This requires methods that allow for medium-to-high throughput compound profiling and that generate quantitative results suitable to establish structure–activity-relationships during lead optimization programs.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

High content analysis of differentiation and cell death in human adipocytes

High content analysis of differentiation and cell death in human adipocytes | High Content Screening | Scoop.it

Understanding adipocyte biology and its homeostasis is in the focus of current obesity research. We aimed to introduce a high-content analysis procedure for directly visualizing and quantifying adipogenesis and adipoapoptosis by laser scanning cytometry (LSC) in a large population of cell. Slide-based image cytometry and image processing algorithms were used and optimized for high-throughput analysis of differentiating cells and apoptotic processes in cell culture at high confluence. Both preadipocytes and adipocytes were simultaneously scrutinized for lipid accumulation, texture properties, nuclear condensation, and DNA fragmentation. Adipocyte commitment was found after incubation in adipogenic medium for 3 days identified by lipid droplet formation and increased light absorption, while terminal differentiation of adipocytes occurred throughout day 9–14 with characteristic nuclear shrinkage, eccentric nuclei localization, chromatin condensation, and massive lipid deposition. Preadipocytes were shown to be more prone to tumor necrosis factor alpha (TNFα)-induced apoptosis compared to mature adipocytes. Importantly, spontaneous DNA fragmentation was observed at early stage when adipocyte commitment occurs. This DNA damage was independent from either spontaneous or induced apoptosis and probably was part of the differentiation program. © 2013 International Society for Advancement of Cytometry

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Scientists develop single-cell imaging method to watch cell clock

Scientists develop single-cell imaging method to watch cell clock | High Content Screening | Scoop.it

A new way to visualize single-cell activity in living zebrafish embryos has allowed scientists to clarify how cells line up in the right place at the right time to receive signals about the next phase of their life.

more...
No comment yet.
Rescooped by CYTOO from Research Workshop
Scoop.it!

Changes in cell shape may lead to metastasis, not the other way around

Changes in cell shape may lead to metastasis, not the other way around | High Content Screening | Scoop.it

A crucial step toward skin cancer may be changes in the genes that control cell shape, report a team of scientists from The Methodist Hospital Research Institute, the Institute of Cancer Research, London, and Harvard Medical School in an upcoming issue of Nature Cell Biology.

 

Using automated high content screening and sophisticated computational modeling, the researchers' screening and analysis of tens of millions of genetically manipulated cells helped them identify more than a dozen genes that influence cell shape. Their work could lead to a better understanding of how cells become metastatic and, eventually, pinpoint new gene therapy targets for cancer treatment.

 

 


Via Jocelyn Stoller
more...
No comment yet.
Scooped by CYTOO
Scoop.it!

A High-Content Screening Assay for Small-Molecule Modulators of Oncogene-Induced Senescence

Cellular senescence is a state of stable cell growth arrest. Activation of oncogenes such as RAS in mammalian cells typically triggers cellular senescence. Oncogene-induced senescence (OIS) is an important tumor suppression mechanism, and suppression of OIS contributes to cell transformation. Oncogenes trigger senescence through a multitude of incompletely understood downstream signaling events that frequently involve protein kinases. To identify target proteins required for RAS-induced senescence, we developed a small-molecule screen in primary human fibroblasts undergoing senescence induced by oncogenic RAS (H-RasG12V). Using a high-content imaging system to monitor two hallmarks of senescence, senescence-associated β-galactosidase activity expression and inhibition of proliferation, we screened a library of known small-molecule kinase inhibitors for those that suppressed OIS. Identified compounds were subsequently validated and confirmed using a third marker of senescence, senescence-associated heterochromatin foci. In summary, we have established a novel high-content screening platform that may be useful for elucidating signaling pathways mediating OIS by targeting critical pathway components.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

A Simple High-Content Cell Cycle Assay Reveals Frequent Discrepancies between Cell Number and ATP and MTS Proliferation Assays

A Simple High-Content Cell Cycle Assay Reveals Frequent Discrepancies between Cell Number and ATP and MTS Proliferation Assays | High Content Screening | Scoop.it

In order to efficiently characterize both antiproliferative potency and mechanism of action of small molecules targeting the cell cycle, we developed a high-throughput image-based assay to determine cell number and cell cycle phase distribution. Using this we profiled the effects of experimental and approved anti-cancer agents with a range mechanisms of action on a set of cell lines, comparing direct cell counting versus two metabolism-based cell viability/proliferation assay formats, ATP-dependent bioluminescence, MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carbo xymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)reduction, and a whole-well DNA-binding dye fluorescence assay. We show that, depending on compound mechanisms of action, the metabolism-based proxy assays are frequently prone to 1) significant underestimation of compound potency and efficacy, and 2) non-monotonic dose-response curves due to concentration-dependent phenotypic ‘switching’. In particular, potency and efficacy of DNA synthesis-targeting agents such as gemcitabine and etoposide could be profoundly underestimated by ATP and MTS-reduction assays. In the same image-based assay we showed that drug-induced increases in ATP content were associated with increased cell size and proportionate increases in mitochondrial content and respiratory flux concomitant with cell cycle arrest. Therefore, differences in compound mechanism of action and cell line-specific responses can yield significantly misleading results when using ATP or tetrazolium-reduction assays as a proxy for cell number when screening compounds for antiproliferative activity or profiling panels of cell lines for drug sensitivity.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Actin in Action: Imaging Approaches to Study Cytoskeleton Structure and Function

Actin in Action: Imaging Approaches to Study Cytoskeleton Structure and Function | High Content Screening | Scoop.it
The cytoskeleton plays several fundamental roles in the cell, including organizing the spatial arrangement of subcellular organelles, regulating cell dynamics and motility, providing a platform for interaction with neighboring cells, and ultimately defining overall cell shape. Fluorescence imaging has proved to be vital in furthering our understanding of the cytoskeleton, and is now a mainstay technique used widely by cell biologists. In this review we provide an introduction to various imaging modalities used to study focal adhesions and the actin cytoskeleton, and using specific examples we highlight a number of recent studies in animal cells that have advanced our knowledge of cytoskeletal behavior.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

HCS-Neurons: identifying phenotypic changes in multi-neuron images upon drug treatments of high-content screening

HCS-Neurons: identifying phenotypic changes in multi-neuron images upon drug treatments of high-content screening | High Content Screening | Scoop.it
Background

High-content screening (HCS) has become a powerful tool for drug discovery. However, the discovery of drugs targeting neurons is still hampered by the inability to accurately identify and quantify the phenotypic changes of multiple neurons in a single image (named multi-neuron image) of a high-content screen. Therefore, it is desirable to develop an automated image analysis method for analyzing multi-neuron images.

Results

We propose an automated analysis method with novel descriptors of neuromorphology features for analyzing HCS-based multi-neuron images, called HCS-neurons. To observe multiple phenotypic changes of neurons, we propose two kinds of descriptors which are neuron feature descriptor (NFD) of 13 neuromorphology features, e.g., neurite length, and generic feature descriptors (GFDs), e.g., Haralick texture. HCS-neurons can 1) automatically extract all quantitative phenotype features in both NFD and GFDs, 2) identify statistically significant phenotypic changes upon drug treatments using ANOVA and regression analysis, and 3) generate an accurate classifier to group neurons treated by different drug concentrations using support vector machine and an intelligent feature selection method. To evaluate HCS-neurons, we treated P19 neurons with nocodazole (a microtubule depolymerizing drug which has been shown to impair neurite development) at six concentrations ranging from 0 to 1000 ng/mL. The experimental results show that all the 13 features of NFD have statistically significant difference with respect to changes in various levels of nocodazole drug concentrations (NDC) and the phenotypic changes of neurites were consistent to the known effect of nocodazole in promoting neurite retraction. Three identified features, total neurite length, average neurite length, and average neurite area were able to achieve an independent test accuracy of 90.28% for the six-dosage classification problem. This NFD module and neuron image datasets are provided as a freely downloadable MatLab project at http://iclab.life.nctu.edu.tw/HCS-Neurons webcite.

Conclusions

Few automatic methods focus on analyzing multi-neuron images collected from HCS used in drug discovery. We provided an automatic HCS-based method for generating accurate classifiers to classify neurons based on their phenotypic changes upon drug treatments. The proposed HCS-neurons method is helpful in identifying and classifying chemical or biological molecules that alter the morphology of a group of neurons in HCS.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Super-Resolution Imaging Strategies for Cell Biologists Using a Spinning Disk Microscope

Super-Resolution Imaging Strategies for Cell Biologists Using a Spinning Disk Microscope | High Content Screening | Scoop.it

In this study we use a spinning disk confocal microscope (SD) to generate super-resolution images of multiple cellular features from any plane in the cell. We obtain super-resolution images by using stochastic intensity fluctuations of biological probes, combining Photoactivation Light-Microscopy (PALM)/Stochastic Optical Reconstruction Microscopy (STORM) methodologies. We compared different image analysis algorithms for processing super-resolution data to identify the most suitable for analysis of particular cell structures. SOFI was chosen for X and Y and was able to achieve a resolution of ca. 80 nm; however higher resolution was possible >30 nm, dependant on the super-resolution image analysis algorithm used. Our method uses low laser power and fluorescent probes which are available either commercially or through the scientific community, and therefore it is gentle enough for biological imaging. Through comparative studies with structured illumination microscopy (SIM) and widefield epifluorescence imaging we identified that our methodology was advantageous for imaging cellular structures which are not immediately at the cell-substrate interface, which include the nuclear architecture and mitochondria. We have shown that it was possible to obtain two coloured images, which highlights the potential this technique has for high-content screening, imaging of multiple epitopes and live cell imaging.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Cell-Based Fuzzy Metrics Enhance High-Content Screening (HCS) Assay Robustness

High-content screening (HCS) allows the exploration of complex cellular phenotypes by automated microscopy and is increasingly being adopted for small interfering RNA genomic screening and phenotypic drug discovery. We introduce a series of cell-based evaluation metrics that have been implemented and validated in a mono-parametric HCS for regulators of the membrane trafficking protein caveolin 1 (CAV1) and have also proved useful for the development of a multiparametric phenotypic HCS for regulators of cytoskeletal reorganization. Imaging metrics evaluate imaging quality such as staining and focus, whereas cell biology metrics are fuzzy logic–based evaluators describing complex biological parameters such as sparseness, confluency, and spreading. The evaluation metrics were implemented in a data-mining pipeline, which first filters out cells that do not pass a quality criterion based on imaging metrics and then uses cell biology metrics to stratify cell samples to allow further analysis of homogeneous cell populations. Use of these metrics significantly improved the robustness of the monoparametric assay tested, as revealed by an increase in Z′ factor, Kolmogorov-Smirnov distance, and strict standard mean difference. Cell biology evaluation metrics were also implemented in a novel supervised learning classification method that combines them with phenotypic features in a statistical model that exceeded conventional classification methods, thus improving multiparametric phenotypic assay sensitivity.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

A High-Content Screen Identifies Novel Compounds That Inhibit Stress-Induced TDP-43 Cellular Aggregation and Associated Cytotoxicity

TDP-43 is an RNA binding protein found to accumulate in the cytoplasm of brain and spinal cord from patients affected with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Nuclear TDP-43 protein regulates transcription through several mechanisms, and under stressed conditions, it forms cytoplasmic aggregates that co-localize with stress granule (SG) proteins in cell culture. These granules are also found in the brain and spinal cord of patients affected with ALS and FTLD. The mechanism through which TDP-43 might contribute to neurodegenerative diseases is poorly understood. To investigate the pathophysiology of TDP-43 aggregation and to isolate potential therapeutic targets, we screened a chemical library of 75,000 compounds using high-content analysis with PC12 cells that inducibly express human TDP-43 tagged with green fluorescent protein (GFP). The screen identified 16 compounds that dose-dependently decreased the TDP-43 inclusions without significant cellular toxicity or changes in total TDP-43 expression levels. To validate the effect, we tested compounds by Western blot analysis and in a Caenorhabditis elegans model that replicates some of the relevant disease phenotypes. The hits from this assay will be useful for elucidating regulation of TDP-43, stress granule response, and possible ALS therapeutics.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

A High Content Assay to Assess Cellular Fitness

A universal process in experimental biology is the use of engineered cells; more often, stably or transiently transfected cells are generated for the purpose. Therefore, it is important that cell health assessment is conducted to check for stress mediated by induction of heat shock proteins (Hsps). For this purpose, we have developed an integrated platform that would enable a direct assessment of transfection efficiency (TE) combined with cellular toxicity and stress response. We make use of automated microscopy and high content analysis to extract from the same well a multiplexed readout to assess and determine optimal chemical transfection conditions. As a proof of concept, we investigated seven commercial reagents, in a matrix of dose and time, to study transfection of an EGFP DNA plasmid into HeLa cells and their consequences on health and fitness; where we scored for cellular proliferation, EGFP positive cells, and induction of Hsp10 and Hsp70 as makers of stress responses. FuGENE HD emerged as the most optimal reagent with no apparent side effects suitable for performing microtiter based miniaturized transfection for both chemical and RNAi screening. In summary, we report on a high content assay method to assess cellular overall fitness upon chemical transfection.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Understanding the impact of image quality on segmentation accuracy

Understanding the impact of image quality on segmentation accuracy | High Content Screening | Scoop.it

High-content screening (HCS) is an automated microscopy technique that enables evaluation of spatial and temporal effects on cells for drug discovery and other applications. The throughput of HCS can be on the order of hundreds of cell images per second to capture transient morphological (structural) effects. Higher throughput can have an impact on image quality, for instance, by reducing exposure times. It can also affect the accuracy of image post-processing (such as the classification of pixels into biologically meaningful regions, known as segmentation). The motivation for our work is to understand the triangular relationship between image quality, microscopy settings, and accuracy of image post-processing.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Quantitative single cell and single molecule proteomics for clinical studies

Quantitative single cell and single molecule proteomics for clinical studies | High Content Screening | Scoop.it

A central aspect of cellular systems biology is the study of cell-to-cell variability driven by network control of molecular noise. Proteins are produced in stochastic bursts and, although time averaging smoothes their accumulated levels, variation in their copy number is substantial in members of environmental sensing and signalling networks. We have developed a label-free, microfluidic antibody capture chip platform called the MAC chip, to quantify precisely the copy numbers of many proteins from a single cell in a multiplexed single assay format. We intend to investigate protein noise in circulating tumour cells (CTCs) isolated from biopsies of cancer patients through the identification of biomolecular signatures, such as p53 tumour suppressor protein, which correlate with biological properties and clinical outcomes during treatment.

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Fluorescence imaging of flowing cells using a temporally coded excitation

Fluorescence imaging of flowing cells using a temporally coded excitation | High Content Screening | Scoop.it

Imaging fluorescence in moving cells is fundamentally challenging because the exposure time is constrained by motion-blur, which limits the available signal. We report a method to image fluorescently labeled leukemia cells in fluid flow that has an effective exposure time of up to 50 times the motion-blur limit. Flowing cells are illuminated with a pseudo-random excitation pulse sequence, resulting in a motion-blur that can be computationally removed to produce near diffraction-limited images. This method enables observation of cellular organelles and their behavior in a fluid environment that resembles the vasculature.

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

What Are High Content Analysis And Content Screening?

In the world of medical science, breakthroughs are happening every moment. Almost every ailment in the world now has a cure, giving afflicted patients all around the world a hope to live the life as they would want it.

 

 

more...
No comment yet.
Scooped by CYTOO
Scoop.it!

Discovery of autophagy modulators through the construction of a high-content screening platform via monitoring of lipid droplets

Given its importance in various cellular processes, autophagy has attracted growing attention as a promising therapeutic target in many human diseases. Herein, we report the construction of a high-content screening platform for the discovery of autophagy modulators via monitoring of cellular lipid droplets as a late-stage marker of autophagy. Notably, our platform discriminates the effects of chemical modulators on autophagic flux without additional transfection, washing, or fixation. This approach allows the simultaneous identification of completely different types of autophagy regulators in a single operation. Mode-of-action studies of 2 representative hit compounds confirmed 1 compound as an mTOR-independent autophagy activator and the other as an autophagy inhibitor that blocks autophagic flux.

 

 

more...
No comment yet.