What is 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. The ability to study cellular and subcellular structure, function, and organization in living systems aids in the development of assays that are more biologically relevant and that can better predict the human response to new drug candidates. 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.
Benefits of live cell imaging
Live cell imaging is a fundamental research tool in cell biology labs and in a wide variety of industries, and it has led to the discovery of drug targets and candidates as well as the molecular mechanisms involved in diseases. Critical for these researchers, is the ability to develop assays that can best mimic the in vivo nature of cells. Benefits of imaging live cells include:
- Monitoring live cells over time provides more than a snapshot of what is occurring in the cells while allowing for the visualization of transient events that may be missed in end-point assays and revealing optimal time-points for end-point assays.
- Tight regulation of environmental conditions ensures that the cellular structures and processes being studied are in their native physiological state
- Imaging live cells in their natural state minimizes artifacts that can arise from other imaging techniques, such as cell fixation and immunostaining.
- The localization and transport of cellular biomolecules along with the progression of multiple pathways can be investigated simultaneously in real-time.
- Acquiring images laterally, axially, and temporally enables the generation of 4D images and data
- Live cell imaging of molecular dynamics generates qualitative and quantitative data that can’t be gained from other biochemical methods.
Cellular imaging and analysis workflow
Protocols for culturing, plating, and maintaining live cells will vary between different cell models however, the steps below outlines a generalized workflow for live cell imaging assays.
Plate cells into labware of choice (chamber slides, dishes, microplates)
Treat cells – Perform cell treatments with desired compounds, RNAi, etc. if part of the assay workflow.
Stain for markers – If required, label cells with the desired fluorophores (fluorescent dyes, fluorescent protein – peptide fusions, etc.
Environmental controls – If the acquisition will take a long time or if the assay is a live cell time-lapse experiment (continuous or discontinuous), then enable full environmental control including gas, temperature, and humidity.
Acquire cell image – Place the plate into the automated 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
Live cell imaging system
Sophisticated live cell imaging instruments provide the ability to regulate environmental conditions to maintain the health and viability of cells while monitoring molecular and cellular dynamics from the single cell to organismal level. From the study of fast-kinetic events to running long-term, time-lapse assays, automated live cell imaging provides the direct observation of dynamic biological processes and the ability to generate a wealth of cellular information
- Label-free, brightfield imaging to multi-color fluorescence imaging capabilities provide the flexibility necessary for the multitude of live cell imaging applications
- High-acquisition speeds allow for the examination of fast-kinetic events, such as calcium oscillation in cardiomyocytes.
- Robust focusing capabilities of automated live cell imaging instruments ensure the maintenance of optimum focus through the entirety of the acquisition, which is crucial for generating accurate live cell imaging data.
Applications and assays
Advancements in automated microscopes with integrated environmental control chambers as well as in fluorescent protein and synthetic fluorophore technology have expanded the capabilities of researchers investigating biological processes in living cells.
我们的高内涵成像和分析系统有灵活的可扩展性，可轻松随着您研究的推进改进您的系统。They feature options and modules to address your specific research needs including objectives, filters, imaging modes, and environmental control. Here we have a collection of applications and assays that utilize these key features for your live cell imaging research:
Live Cell Imaging and Analysis Workflow
The described live cell imaging workflow is a generalized example of a continuous long-term time-lapse live cell assay. This workflow breaks down the steps from cell plating to image analysis and highlights the materials and instrumentation necessary to run a live cell assay including an automated high-content imaging system and cellular imaging and analysis software.
Plate adherent or suspension cells into the labware of choice including culture dishes, chamber slides, and microplates. Incubate the cells at normal culture conditions (i.e. 37°C, 5% CO2). For adherent cells, incubating the plated cells overnight will allow ample time for cell attachment and growth.
96 或 384 孔板
Most cell cultures for high-content, high-throughput screening are run using 96- or 384-well microplates.
If necessary for the protocol, treat the cells with compounds of interest.
Compounds can be added to cells for minutes up to several days depending on the mechanism of action of the compounds and the biological re-sponse being investigated. For example, apoptosis or receptor internalization assays generally require shorter incubation periods while multiparametric cytotoxicity assays require longer periods. Longer compound treatments may require the replacement of compounds during incubation
Live cells can be stained with a variety of fluorophores or transfected/transduced with constructs containing fluorescent protein – peptide fusions. Manufacturer instructions should be followed when staining. Stain-ing isn’t necessary for all live cell applications and label free imaging and analysis allows for the tracking and monitoring of cells in brightfield only.
简单易用、可靠的检测试剂盒，适用于生命科学研究、药物的探索与发现以及生物学测定。我们的检测试剂盒已经过优化，可用于我们的仪器。Screen more compounds earlier in drug discovery and enable characterization of a full concentra-tion-response profile of test compounds.商店分析试剂盒
ImageXpress High-content Imaging Portfolio
我们的高内涵成像和分析系统有灵活的可扩展性，可轻松随着您研究的推进改进您的系统。They feature options and modules to address your specific research including objectives, filters, imaging modes, and envi-ronmental conditions.View product comparison table
CONFIGURE ENVIRONMENTAL CONTROL SETTINGS
After the addition of stains and compounds, place the plate into the appropriate imaging instrument integrated with environmental control capabilities such asl gases (CO2, O2), temperature, and humidity.
For long-term time-lapse assays it is essential to maintain a natural physiological environment to ensure the health of living cells and to prevent focus drift throughout the time-lapse acquisition.
ACQUIRE LIVE CELL IMAGE
For continuous time-lapse acquisitions, configure the total length of time for the time series and the imaging intervals, i.e. image every hour for 24 hours.
Discontinuous time-lapse acquisitions can be per-formed, where the plate is taken to and from the incubator and imaged at different time points.
ImageXpress® Pico 个人型高内涵成像分析系统不仅仅是一台数字显微镜，还结合了高分辨率成像及强大的分析功能。Whether running fluorescence imaging or brightfield assays, the automated imager features comprehensive preconfigured protocols for cell-based assays to shorten the learning curve, so you can start running experiments quickly.查看产品
Cellular Imaging Acquisition and Analysis Software
The CellReporterXpress Automated Image Acquisition and Analysis Software works with the ImageXpress® Pico systems. It has a clean, easy-to-learn interface for performing quantitative analysis on images acquired from automated microscopy. 该软件可实现在更高通量下对图像进行分布式分析，非常适用于对玻片或微孔板进行缩放式显微成像。查看产品
ANALYZE LIVE CELL IMAGE
Analyze the images with cellular imaging analysis software to generate multiparametric readouts for the biological responses that are being studied. Kinetic data can be generated and reviewed for time-lapse acquisitions.
Resources for Live Cell Imaging
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High-content assay for morphological characterization of 3D neuronal networks in a microfluidic platform
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