Imaging Analysis & Live Cell Imaging
Live-cell imaging and analysis allows scientists to study dynamic cellular events in real time to gain unique insights into biology. These techniques are in contrast to PCR, flow cytometry, and immunocytochemistry and tissue staining with antibodies, which analyze cellular events as a snapshot. Today, improvements in cameras, live-cell fluorescent dyes, fluorescent proteins, video compression, and time-lapse microscopy have advanced our ability to visualize and analyze living cells, their intercellular interactions, and subcellular processes with remarkable detail and fidelity.
Related Technical Articles
- Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.
- Firefly luciferase is a widely used bioluminescent reporter for studying gene regulation and function. It is a very sensitive genetic reporter due to the absence of endogenous luciferase activity in mammalian cells or tissues.
- PKH and CellVue® Fluorescent Cell Linker Kits provide fluorescent labeling of live cells over an extended period of time, with no apparent toxic effects.
- Fluorescent calcium indicators and dyes to measure Ca+ flux used in calcium imaging experiments. Calcium ions (Ca2+) play vital cellular physiology roles in signal transduction pathways, in neurotransmitter release, in contraction of all muscle cell types, as enzyme cofactors, and in fertilization.
- Lipophilic cell tracking dyes enable cancer biologists to track tumor and immune cell functions both in vitro and in vivo. Read the article to choose a right membrane dye kit for cell tracking and proliferation monitoring.
- See All (32)
Related Protocols
- PKH dyes have been successfully used to label exosomes and extracellular vesicles for in vitro and in vivo tracking experiments. The exosome labeling protocol below details steps for reliable fluorescent labeling of exosomes using PKH dyes for microvesicle tracking experiments.
- Cell staining can be divided into four steps: cell preparation, fixation, application of antibody, and evaluation.
- Immunocytochemistry (ICC) Cell Capture Imaging Reagent and protocol for eliminating wash steps, and is highly effective in applications involving rare cells, low cell count samples, and limited access samples.
- Toluidine blue stains the nuclear material in tissues with a high DNA and RNA content and is used for selectively staining acidic tissue components such as carboxylates, sulfates, and phosphate radicals.
- This protocol covers 3 modes for the microscopic examination of cell samples.
- See All (6)
Find More Articles and Protocols
Live-cell Imaging Systems
A variety of techniques are used to visualize the complex structures and behavior of cells. In static analyses, cells are fixed and stained, which requires killing the cells and therefore capturing a single instant in cellular time. While static techniques are suitable for some applications, they do not provide insight into cellular activities. Dynamic, live-cell approaches facilitate the visualization of structure, behavior or composition in live cells, without traditional staining. These techniques are ideal for applications that must mimic in vivo conditions, as well as for developing predictive assays for drug screening.
Live-cell imaging systems use advanced optics, environmental control, and reagents that permit long-term studies of mammalian, bacterial, and yeast cells. Culture and imaging systems that also employ microfluidics allow precise control of the culture microenvironment, like flow, gas mixture, and temperature, while limiting physical stress to cells. Live-cell imaging and analysis is suited to a range of applications, including hypoxia, cell migration, 3D cell culture, cytoskeletal dynamics and protein trafficking. Advanced systems automate culture conditions, administration and withdrawal of treatments, and imaging intervals to create images that may be presented as still image or video data.
Live-cell Imaging Reagents
Live-cell imaging media and supplements are designed to protect cells from light-induced cellular damage during time-lapse experiments. These reagents are also formulated to have low autofluorescence and photobleaching, which dramatically enhances the quality of fluorescent live-cell images.
A variety of live-cell staining dyes are available to study the dynamic processes of live cells. Live-cell fluorescent organelle dyes permit the selective staining of specific organelles, such as the cell membrane, nucleus, cytoplasm, mitochondria, lysosomes, endoplasmic reticulum (ER), Golgi apparatus, and cytoskeleton proteins, all without increased cytotoxicity. Using organelle dyes as counterstains in live-cell imaging is also useful in functional studies.
Biosensors with GFP and RFP can be used to detect a specific protein as well as the subcellular location of that protein within live cells by either fluorescent microscopy or time-lapse video capture. Lentiviral vector systems are a popular tool for introducing genes and gene products into cells, with many advantages over chemical-based transfection. Beyond organelle detection, cell-permeable live-cell dyes are used for applications such as apoptosis detection, cell viability, cell health, hypoxia, reactive oxygen species studies, calcium indicator imaging, and neural or stem cell experiments.
To continue reading please sign in or create an account.
Don't Have An Account?