Cell imaging: Acquisition and analysis
Researchers have several options in methods for imaging cells, from phase-contrast microscopy that shows intact cells to fluorescent imaging of single molecules or organelles. Cellular analysis is performed to evaluate and measure the current state of cells, such as cell integrity, toxicity, and viability and various other research applications. An integral part of cellular analysis is data collection, analysis, and export into a meaningful and useful format.
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
Setting up acquisition and analysis on the ImageXpress Pico
View our “How-to” video demonstrating the setup and analysis of an acquisition using a digital microscopy system that combines high-resolution imaging with powerful analysis software.
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
The development and integration of three-dimensional (3D) assay models are becoming popular to drive translational biology. Our ImageXpress® Micro Confocal High-Content Imaging System allows superior visualization of thick samples by minimizing background fluorescence and increasing sharpness, resulting in accurate image segmentation. Increase the biological relevance of your screening assays with a seamless integration between acquisition and analysis of cells in a 3D space to yield volume, intensity, and distance measurements.
3D Cell Models
The development and integration of three-dimensional (3D) assay models are becoming popular to drive translational biology. Our ImageXpress® Micro Confocal High-Content Imaging System allows superior visualization of thick samples by minimizing background fluorescence and increasing sharpness, resulting in accurate image segmentation. Increase the biological relevance of your screening assays with a seamless integration between acquisition and analysis of cells in a 3D space to yield volume, intensity, and distance measurements.
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.
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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.
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.
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eBook: Cellular Imaging Insights
Elucidating complex cellular pathways within 2D and 3D cellular structures in response to chemical stimulation or stress is the goal of many researchers. The capacity to which scientists have access to and can adopt emerging cellular imaging and analysis technology fuels their level of understanding.
eBook: Cellular Imaging Insights
Elucidating complex cellular pathways within 2D and 3D cellular structures in response to chemical stimulation or stress is the goal of many researchers. The capacity to which scientists have access to and can adopt emerging cellular imaging and analysis technology fuels their level of understanding.
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.
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 Nano system paired with CellReporterXpress software for analysis has utility in all parts of the stem cells 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 Nano system paired with CellReporterXpress software for analysis has utility in all parts of the stem cells researcher’s workflow from tracking differentiation to quality control to measuring functionality of specific cell types.
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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.
Resources of Cellular Imaging
eBook
Cellular Imaging Insights
Cellular Imaging Insights
Gain insights and expedite studies for 2D and 3D cellular structures using automated cellular imaging.
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…
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
Establishment of physiologically-relevant in vitro models is crucial to further understanding of the mechanisms of neurological diseases as well as targeted drug development. While iPSC-…
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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
There is increasing interest in exploring the use of three-dimensional (3D) spheroids for modeling developmental and tissue biology with the goal of accelerating translational research in…
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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
Cardiotoxicity remains a primary cause for drug attrition during clinical trials and post-marketing. While it is believed that a significant percentage of cardiovascular diseases are due to…
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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…