Cell imaging: Acquisition and analysis
Cellular analysis workflow
Protocols for culturing, plating, and maintaining cells will vary between different cell models however, the steps below outline a general workflow for cell-based imaging assays from upstream media prep through downstream experimental analysis using a high-content imaging system and cellular analysis software.
- Plate cells into labware of choice (chamber slides, dishes, microplates)
- Treat cells – Perform cell treatments with desired compounds if part of the assay workflow
- Stain for markers – If required, label cells with the desired fluorophores (fluorescent dyes, fluorescent protein – peptide fusions, etc.
- Acquire cell image – Place the plate into the cell imaging system and start acquiring your live cell images
- Analyze cell image – Use cellular imaging analysis software to run quantitative analysis of live cell images
Accelerate your screening with high-content and automated imaging
This technical webinar will focus on high-content screening applications such as those involved in viral research, neurite outgrowth studies, and organoid models. The information provided will apply to researchers currently working with cell-based assays and doing live-cell imaging, and those looking to automate and/or increase their current imaging workflow.
Types of cellular imaging methods
Whether the demand is for a high-content and high-throughput or a simple-to-use system, our cellular imaging solutions can accommodate acquisition and analysis that suits the needs of a screening or research lab. Here, we’ve briefly described several methods in cellular imaging:
- Phase-contrast microscopy shows intact cells
- Fluorescent imaging can show single molecules or organelles
- Brightfield microscopy is useful for visualizing whole cells rapidly
- Confocal microscopy is preferred for high-quality images and 3D reconstruction of cells
- 4D live-cell imaging combines confocal with time-lapse microscopy
- Automated microscopy employs software and hardware features which ensure robust focus in every field-of-view combined with sophisticated image analysis software to generate repeatable and subjective quantitative data
Common cell-based assays and applications
Here, we’ve highlighted several cell-based assay topics from 3D cell cultures to stem cell research as well as applications for cardiomyocytes and toxicity screening.
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3D Cell Models
3D cell cultures offer the advantage of closely recapitulating aspects of human tissues including the architecture, cell organization, cell-cell and cell-matrix interactions, and more physiologically-relevant diffusion characteristics. Utilization of 3D cellular assays adds value to research and screening campaigns, spanning the translational gap between 2D cell cultures and whole-animal models. By reproducing important parameters of the in vivo environment, 3D models can provide unique insight into the behavior of stem cells and developing tissues in vitro.
3D Cell Models
3D cell cultures offer the advantage of closely recapitulating aspects of human tissues including the architecture, cell organization, cell-cell and cell-matrix interactions, and more physiologically-relevant diffusion characteristics. Utilization of 3D cellular assays adds value to research and screening campaigns, spanning the translational gap between 2D cell cultures and whole-animal models. By reproducing important parameters of the in vivo environment, 3D models can provide unique insight into the behavior of stem cells and developing tissues in vitro.
Cancer Research
Cancer researchers need tools that enable them to more easily study the complex and often poorly understood interactions between cancerous cells and their environment, and to identify points of therapeutic intervention. Learn about instrumentation and software that facilitate cancer research using, in many cases, biologically relevant 3D cellular models like spheroids, organoids, and organ-on-a-chip systems that simulate the in vivo environment of a tumor or organ.
Cancer Research
Cancer researchers need tools that enable them to more easily study the complex and often poorly understood interactions between cancerous cells and their environment, and to identify points of therapeutic intervention. Learn about instrumentation and software that facilitate cancer research using, in many cases, biologically relevant 3D cellular models like spheroids, organoids, and organ-on-a-chip systems that simulate the in vivo environment of a tumor or organ.
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Cardiomyocytes
Stem cells differentiated into cardiomyocytes are used to screen early for potential toxicological effects of drugs, thus helping to avoid investment in development of drugs which will fail in clinical trials due to cardiac toxicity.
Cardiomyocytes
Stem cells differentiated into cardiomyocytes are used to screen early for potential toxicological effects of drugs, thus helping to avoid investment in development of drugs which will fail in clinical trials due to cardiac toxicity.
Cell Counting
Cell counting is fundamental and critical to numerous biological experiments. Assays such as drug compound toxicity, cell proliferation, and inhibition of cell division rely on the assessment of the number or density of cells in a well.
Cell Counting
Cell counting is fundamental and critical to numerous biological experiments. Assays such as drug compound toxicity, cell proliferation, and inhibition of cell division rely on the assessment of the number or density of cells in a well.
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Cell Migration Assay
The movement or migration of cells is often measured in vitro to elucidate the mechanisms of various physiological activities such as wound healing or cancer cell metastasis. Cell migration assays may be conducted in a controlled environment using live cell timelapse imaging.
Cell Migration Assay
The movement or migration of cells is often measured in vitro to elucidate the mechanisms of various physiological activities such as wound healing or cancer cell metastasis. Cell migration assays may be conducted in a controlled environment using live cell timelapse imaging.
Cell Painting
Cell Painting is a high-content, multiplexed image-based assay used for cytological profiling. In a Cell Painting assay, up to six fluorescent dyes are used to label different components of the cell including the nucleus, the endoplasmic reticulum, mitochondria, cytoskeleton, the Golgi apparatus, and RNA. The goal is to “paint” as much of the cell as possible to capture a representative image of the whole cell.
Cell Painting
Cell Painting is a high-content, multiplexed image-based assay used for cytological profiling. In a Cell Painting assay, up to six fluorescent dyes are used to label different components of the cell including the nucleus, the endoplasmic reticulum, mitochondria, cytoskeleton, the Golgi apparatus, and RNA. The goal is to “paint” as much of the cell as possible to capture a representative image of the whole cell.
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COVID-19 and Infectious Disease Research
Here we've addressed common applications in infectious disease research including cell line development, binding affinity, viral neutralization, viral titer and more with a focused effort on understanding the SARS-CoV-2 virus in order to develop potential therapies for COVID-19 including vaccines, therapeutics and diagnostics.
COVID-19 and Infectious Disease Research
Here we've addressed common applications in infectious disease research including cell line development, binding affinity, viral neutralization, viral titer and more with a focused effort on understanding the SARS-CoV-2 virus in order to develop potential therapies for COVID-19 including vaccines, therapeutics and diagnostics.
Live Cell Imaging
Live cell imaging is the study of cellular structure and function in living cells via microscopy. It enables the visualization and quantitation of dynamic cellular processes in real time. Live cell imaging encompasses a broad range of topics and biological applications—whether it is performing long-term kinetic assays or fluorescently labeling live cells.
Live Cell Imaging
Live cell imaging is the study of cellular structure and function in living cells via microscopy. It enables the visualization and quantitation of dynamic cellular processes in real time. Live cell imaging encompasses a broad range of topics and biological applications—whether it is performing long-term kinetic assays or fluorescently labeling live cells.
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Neurite Outgrowth / Neurite Tracing
Neurons create connections via extensions of their cellular body called processes. This biological phenomenon is referred to as neurite outgrowth. Understanding the signaling mechanisms driving neurite outgrowth provides valuable insight into neurotoxic responses, compound screening, and for interpreting factors influencing neural regeneration. Using the ImageXpress Micro system in combination with MetaXpress Image Analysis Software automated neurite outgrowth imaging and analysis is possible for slide or microplate-based cellular assays.
Neurite Outgrowth / Neurite Tracing
Neurons create connections via extensions of their cellular body called processes. This biological phenomenon is referred to as neurite outgrowth. Understanding the signaling mechanisms driving neurite outgrowth provides valuable insight into neurotoxic responses, compound screening, and for interpreting factors influencing neural regeneration. Using the ImageXpress Micro system in combination with MetaXpress Image Analysis Software automated neurite outgrowth imaging and analysis is possible for slide or microplate-based cellular assays.
Organoid Innovation Center
The new Organoid Innovation Center at Molecular Devices combines cutting-edge technologies with novel 3D biology methods to address key challenges of scaling complex 3D biology.
The collaborative space brings customers and researchers into the lab to test automated workflows for organoid culturing and screening, with guidance from in-house scientists.
Organoid Innovation Center
The new Organoid Innovation Center at Molecular Devices combines cutting-edge technologies with novel 3D biology methods to address key challenges of scaling complex 3D biology.
The collaborative space brings customers and researchers into the lab to test automated workflows for organoid culturing and screening, with guidance from in-house scientists.
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Stem Cell Research
Pluripotent stem cells can be used for studies in developmental biology or differentiated as a source for organ-specific cells and used for live or fixed cell-based assays on slides or in multi-well plates. The ImageXpress system has utility in all parts of the stem cell researcher’s workflow, from tracking differentiation, to quality control, to measuring functionality of specific cell types.
Stem Cell Research
Pluripotent stem cells can be used for studies in developmental biology or differentiated as a source for organ-specific cells and used for live or fixed cell-based assays on slides or in multi-well plates. The ImageXpress system has utility in all parts of the stem cell researcher’s workflow, from tracking differentiation, to quality control, to measuring functionality of specific cell types.
Toxicity Screening
Screening for off-target or toxic effects is very important during the development of new drugs and for the extension of the therapeutic potential of existing molecules. ImageXpress systems are fully integrated hardware and software platforms for automated acquisition and analysis of images for high-throughput cell-based cytotoxicity testing. Configured with optional environmental control, living cell responses or kinetic reactions can be monitored in real time for several days.
Toxicity Screening
Screening for off-target or toxic effects is very important during the development of new drugs and for the extension of the therapeutic potential of existing molecules. ImageXpress systems are fully integrated hardware and software platforms for automated acquisition and analysis of images for high-throughput cell-based cytotoxicity testing. Configured with optional environmental control, living cell responses or kinetic reactions can be monitored in real time for several days.
Latest Resources
Resources of Cellular Imaging
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Image-based phenotypic profiling using Cell Painting in a 3D breast cancer spheroid model
Image-based phenotypic profiling using Cell Painting in a 3D breast cancer spheroid model
Most potential oncology drugs fail the drug development pipeline, despite having promising data for their efficacy in vitro. This further incentivize the need for identifying in vitro models…
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Scientific Poster
Automation for organoid assays – An integrated system with high-content imaging
Automation for organoid assays – An integrated system with high-content imaging
Three-dimensional (3D) cell models that represent various tissues are being successfully used in drug discovery and disease modeling to study complex biological effects and tissue architectu…
eBook
Building complexity in organoid models
Building complexity in organoid models
Building complexity in organoid models | Molecular Devices
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Automated Monitoring of Development and Activity Analysis of iPSC-derived 3D Cerebral Organoids
Automated Monitoring of Development and Activity Analysis of iPSC-derived 3D Cerebral Organoids
The human brain is highly complex which makes it challenging to study both in vitro and in-model organisms. Cultured neurons do not sufficiently recapitulate the three-dimensional (3D)…
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AI enabled phenotypic analysis of label-free intestinal organoids
AI enabled phenotypic analysis of label-free intestinal organoids
The intestinal crypt system is an important part of the intestinal organoid where mature organoids have more complex and numerous crypt structures. Yet, due to their high turnover rate and…
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Application Note
Neurite outgrowth analysis in 3D reconstructed human brain micro-tissues
Neurite outgrowth analysis in 3D reconstructed human brain micro-tissues
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Automation and high content imaging of 3D triple-negative breast cancer patient-derived tumoroids assay for compound screening
Automation and high content imaging of 3D triple-negative breast cancer patient-derived tumoroids assay for compound screening
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3D Immune Cell Migration
3D Immune Cell Migration
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COVID-19 Timeline: Diagnostics, Vaccines, and Therapeutic Antibody Development
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Automation of the organ-on a chip assay: automated culture, imaging and analysis of angiogenesis
Automation of the organ-on a chip assay: automated culture, imaging and analysis of angiogenesis
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Intestinal organoids for automated screening assays. High content imaging and analysis of organoid morphology
Intestinal organoids for automated screening assays. High content imaging and analysis of organoid morphology
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Label-free cell segmentation with IN Carta SINAP application module
Label-free cell segmentation with IN Carta SINAP application module
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Engineering Next-gen Organoids with Automated Lab Workflows at #SLAS2022
Engineering Next-gen Organoids with Automated Lab Workflows at #SLAS2022
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Improving the robustness of Cell Painting with a near-infrared label and advanced image and data analytics
Improving the robustness of Cell Painting with a near-infrared label and advanced image and data analytics
Image-based phenotypic profiling approaches such as the widely used Cell Painting assay use high-content imaging along with multiparametric readouts to study biological, genetic, and…
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Lab Automation 101: Inside Access From Our Subject Matter Expert
Lab Automation 101: Inside Access From Our Subject Matter Expert
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Application Note
Deep Learning-based Image Analysis for Label-free Live Monitoring of iPSC and 3D Organoid Cultures
Deep Learning-based Image Analysis for Label-free Live Monitoring of iPSC and 3D Organoid Cultures
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Innovation at Molecular Devices: Updates in Automated, High-Content Imaging
Innovation at Molecular Devices: Updates in Automated, High-Content Imaging
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Organoids for disease modeling and in vitro drug screening
Organoids for disease modeling and in vitro drug screening
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Monitoring organoid development and characterization of calcium oscillation activities in iPSC-derived 3D cerebral organoids
Monitoring organoid development and characterization of calcium oscillation activities in iPSC-derived 3D cerebral organoids
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High-throughput assessment of compound-induced pro-arrhythmic effects in human IPSC-derived cardiomyocytes
High-throughput assessment of compound-induced pro-arrhythmic effects in human IPSC-derived cardiomyocytes
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Deep learning-based image analysis for label-free live monitoring of iPSC 3D organoid cultures
Deep learning-based image analysis for label-free live monitoring of iPSC 3D organoid cultures
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Project Profile: Organoid Innovation Center
Project Profile: Organoid Innovation Center
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Tips for running a successful live cell imaging experiment
Tips for running a successful live cell imaging experiment
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Shaping the Future of Organoid Research
Shaping the Future of Organoid Research
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How AI is advancing neurodegenerative disease research
How AI is advancing neurodegenerative disease research
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Disease Modeling with 3D Cell-Based Assays Using a Novel Flowchip System and High-Content Imaging
Disease Modeling with 3D Cell-Based Assays Using a Novel Flowchip System and High-Content Imaging
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Danaher’s SCIEX and Molecular Devices Businesses Debut New Products
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eBook
Cellular Imaging Insights
Cellular Imaging Insights
Gain insights and expedite studies for 2D and 3D cellular structures using automated cellular imaging.
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Advanced Cell Screening and Imaging Reveals How the Cardiac System Responds in Drug Development
Advanced Cell Screening and Imaging Reveals How the Cardiac System Responds in Drug Development
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AI-based analysis of complex biological phenotypes
AI-based analysis of complex biological phenotypes
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Improving acquisition and analysis of 3D cell model assays with water immersion objectives
Improving acquisition and analysis of 3D cell model assays with water immersion objectives
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The workflow game-changer: combined technologies for efficiency
The workflow game-changer: combined technologies for efficiency
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Neuroscience: bridging the gap between cell-based and human research
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Imaging system produces human cellular morphological data for COVID-19 research
Imaging system produces human cellular morphological data for COVID-19 research
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From 3D high throughput to simple automated imaging: Cellular imaging within reach of every laboratory
From 3D high throughput to simple automated imaging: Cellular imaging within reach of every laboratory
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What 3D imaging means for cancer drug discovery
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Making the move from 2D to 3D
Making the move from 2D to 3D
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Revolutionizing early drug discovery for immuno-oncology and neurodegenerative disease modeling
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LabTube Meets Molecular Devices & MIMETAS
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High-content screening of complex physiologically-relevant cell models
High-content screening of complex physiologically-relevant cell models
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Multi-Dimensional in Vitro Bioactivity Profiling for Grouping of Glycol Ethers
Multi-Dimensional in Vitro Bioactivity Profiling for Grouping of Glycol Ethers
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Rising growth of the global high‑content screening software and services market
Rising growth of the global high‑content screening software and services market
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Application Note
Assessment of neurotoxicity and neuronal development using induced pluripotent stem cell-based neurite outgrowth assay
Assessment of neurotoxicity and neuronal development using induced pluripotent stem cell-based neurite outgrowth assay
Growing concerns about the increased prevalence of untested chemicals in the environment has created a pressing need to develop reliable and efficient screening tools to identify chemicals…
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Scientific Poster
Customizing Your High Content Analysis for Accelerated Toxicity Screening
Customizing Your High Content Analysis for Accelerated Toxicity Screening
Combining high-throughput imaging and customizable high-content analysis allows multiple useful applications for toxicity assessment in assays using either cell lines or stem-cell derived hu…
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Application Note
Measuring mitochondrial shape and distribution using the ImageXpress Micro High-Content Imaging System
Measuring mitochondrial shape and distribution using the ImageXpress Micro High-Content Imaging System
Using high-content imaging assays allow cells cultured and treated in microplates to be imaged alive without wash steps for real-time studies of cells in their most natural state, although…
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Application Note
High-content assay for morphological characterization of 3D neuronal networks in a microfluidic platform
High-content assay for morphological characterization of 3D neuronal networks in a microfluidic platform
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Application Note
High-content 3D toxicity assay using iPSC-derived hepatocyte spheroids
High-content 3D toxicity assay using iPSC-derived hepatocyte spheroids
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Application Note
High-Throughput Confocal Imaging of Spheroids for Screening Cancer Therapeutics
High-Throughput Confocal Imaging of Spheroids for Screening Cancer Therapeutics
In recent years, there has been significant progress in development of in vitro aggregates of tumor cells for use as models for in vivo tissue environments. When seeded into a well of a…
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Application Note
3D Imaging of Cancer Cell Spheroids
3D Imaging of Cancer Cell Spheroids
Many cancer cell lines will form spheroids if cultured on a favorable three dimensional (3D) matrix. These spheroids are believed to represent tumor physiology more closely than cells…
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Application Note
Phenotypic characterization of toxic compound effects on cardiomyocytes derived from induced pluripotent stem cells
Phenotypic characterization of toxic compound effects on cardiomyocytes derived from induced pluripotent stem cells
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Application Note
Rapid selection of adherent cell lines using ClonePix Technology
Rapid selection of adherent cell lines using ClonePix Technology
During the last decade, production of monoclonal antibodies and recombinant proteins by mammalian cells has become a cutting edge technology. However, the generation of transient as well as…
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Application Note
StainFree technology for cellular image analysis without fluorescent dyes
StainFree technology for cellular image analysis without fluorescent dyes
Imaging cell-based assays typically requires the use of fluorescent probes that can be toxic to living cells or may only function in fixed cells. A label-free method for analyzing cell…
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Application Note
Live cell kinetics assay utilizing the ImageXpress Micro System
Live cell kinetics assay utilizing the ImageXpress Micro System
The ImageXpress® Micro Confocal/ XLS Widefield High-Content Imaging System from Molecular Devices includes optional Fluidics and Environmental Control modules, enabling single-channel…
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Application Note
High-content screening of neuronal toxicity using iPSC-derived human neurons
High-content screening of neuronal toxicity using iPSC-derived human neurons
The nervous system is sensitive to the toxic effects of many chemical compounds, environmental agents and certain naturally occurring substances. Neurotoxicity can cause temporary or…
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Application Note
Multiplexed high-content hepatotoxicity assays using iPSC-derived hepatocytes
Multiplexed high-content hepatotoxicity assays using iPSC-derived hepatocytes
Drug-induced hepatotoxicity is an important cause for liver injury and acute liver failure. Thus highly predictive assays for safety and efficacy testing are crucial for improving drug…
Data Sheet
AcuityXpress High Content Informatics Software
AcuityXpress High Content Informatics Software
AcuityXpress™ Software provides cellular informatics for the Complete Imaging Solution from Molecular Devices and has been specifically designed to address the needs of high-content data ana…
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Publications
Phenotypic Assays for Characterizing Compound Effects on Induced Pluripotent Stem Cell-Derived Cardiac Spheroids
Phenotypic Assays for Characterizing Compound Effects on Induced Pluripotent Stem Cell-Derived Cardiac Spheroids
Development of more complex, biologically relevant, and predictive cell-based assays for compound screening is a major challenge in drug discovery. The focus of this study was to establish h…
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Publications
In Vitro Cardiotoxicity Assessment of Environmental Chemicals Using an Organotypic Human Induced Pluripotent Stem Cell-Derived Model
In Vitro Cardiotoxicity Assessment of Environmental Chemicals Using an Organotypic Human Induced Pluripotent Stem Cell-Derived Model
An important target area for addressing data gaps through in vitro screening is the detection of potential cardiotoxicants. Despite the fact that current conservative estimates relate at lea…
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Publications
Determination of Hepatotoxicity in iPSC-Derived Hepatocytes by Multiplexed High Content Assays
Determination of Hepatotoxicity in iPSC-Derived Hepatocytes by Multiplexed High Content Assays
We present here methods for assessing hepatotoxicity by high content imaging and image analysis. The assays focus on the characterization of toxic effects using a variety of phenotypic endpo…
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Publications
Phenotypic Characterization of Toxic Compound Effects on Liver Spheroids Derived from iPSC Using Confocal Imaging and Three-Dimensional Image Analysis
Phenotypic Characterization of Toxic Compound Effects on Liver Spheroids Derived from iPSC Using Confocal Imaging and Three-Dimensional Image Analysis
Cell models are becoming more complex to better mimic the in vivo environment and provide greater predictivity for compound efficacy and toxicity. There is an increasing interest in explorin…
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Publications
A Chemical-Biological Similarity-Based Grouping of Complex Substances as a Prototype Approach for Evaluating Chemical Alternatives
A Chemical-Biological Similarity-Based Grouping of Complex Substances as a Prototype Approach for Evaluating Chemical Alternatives
Comparative assessment of potential human health impacts is a critical step in evaluating both chemical alternatives and existing products on the market. Most alternatives assessments are co…
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Publications
Neurite Outgrowth in Human Induced Pluripotent Stem Cell-Derived Neurons as a High-Throughput Screen for Developmental Neurotoxicity or Neurotoxicity
Neurite Outgrowth in Human Induced Pluripotent Stem Cell-Derived Neurons as a High-Throughput Screen for Developmental Neurotoxicity or Neurotoxicity
Due to the increasing prevalence of neurological disorders and the large number of untested compounds in the environment, there is a need to develop reliable and efficient screening tools to…
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Publications
High-Content Assay Multiplexing for Toxicity Screening in Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Hepatocytes
High-Content Assay Multiplexing for Toxicity Screening in Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Hepatocytes
Cell-based high-content screening (HCS) assays have become an increasingly attractive alternative to traditional in vitro and in vivo testing in pharmaceutical drug development and toxicolog…
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Publications
High-Content Assays for Characterizing the Viability and Morphology of 3D Cancer Spheroid Cultures
High-Content Assays for Characterizing the Viability and Morphology of 3D Cancer Spheroid Cultures
There is an increasing interest in using three-dimensional (3D) spheroids for modeling cancer and tissue biology to accelerate translation research. Development of higher throughput assays t…
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Publications
Abstract 5099: Automated rapid and accurate cell cycle analysis
Abstract 5099: Automated rapid and accurate cell cycle analysis
It is essential to have detailed information about compounds identified in screening campaigns during drug discovery and to know their influence on cell cycle progression or DNA repair. The…
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Publications
High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells
High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells
Abstract Development of predictive in vitro assays for early toxicity evaluation is extremely important for improving the drug development process and reducing drug attrition rates during cl…
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Publications
Assessment of Beating Paramaters in Human Induced Pluripotent Stem Cells Enables Quantitative in Vitro Screening for Cadiotoxicity
Assessment of Beating Paramaters in Human Induced Pluripotent Stem Cells Enables Quantitative in Vitro Screening for Cadiotoxicity
Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes show promise for screening during early drug development. Here, we tested a hypothesis that in vitro assessment of multiple…
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Publications
Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
A large percentage of drugs fail in clinical studies due to cardiac toxicity; thus, development of sensitive in vitro assays that can evaluate potential adverse effects on cardiomyocytes is…
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High-Content Phenotypic Screening
ISSCR 2021 Innovation Showcase: Automated Culture and High-Content Imaging of 3D Lung and Cardiac
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Transitioning high-content assays to 3D: Scientific opportunities and imaging challenges
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