Live cell imaging is a rapidly evolving field that involves the visual observation of living cells over time under controlled conditions. The technique is a powerful solution to investigate cellular processes, dynamics, and functions in real-time, paving the way for numerous breakthroughs in the scientific and medical fields. Live cell imaging allows the observation of cells in their native and functioning state, providing a more realistic picture of cellular processes than static imaging of fixed cells. This non-invasive technique is suitable for observing cell growth in vitro under culture conditions.
Three technically critical factors for live cell imaging are minimizing phototoxicity and photobleaching, maintaining cell viability during the imaging process, and obtaining adequate resolution and contrast. Prolonged exposure to light or certain wavelengths can damage cells (phototoxicity) or reduce the fluorescence of dyes (photobleaching). Therefore, careful selection of light sources, dyes, and imaging parameters is critical. During the imaging process, the cells must be maintained in an optimal environment, such as the correct temperature, pH, and gas mixture, to ensure proper cell function. To visualize specific cellular structures or processes, high-resolution imaging with good contrast is necessary. The selection of an appropriate microscope, imaging modality, and sample preparation technique is essential.
Advanced live cell imaging systems are capable of regulating environmental conditions to maintain cell health and viability while monitoring molecular and cellular dynamics from the single cell to the organism level. Many types of live cell imaging systems have unique advantages and are designed for different applications. Label-free, bright-field imaging to multi-color fluorescence imaging capabilities provide the flexibility necessary for a wide range of live cell imaging applications. Fluorescence microscopy is the most commonly used live cell imaging system. This system is suitable for studying intracellular molecular interactions, protein localization, and cellular signaling pathways. Confocal microscopy is a live-cell imaging system that uses spot illumination and a pinhole in an optically conjugate plane in front of the detector to eliminate out-of-focus signals. Confocal microscopy is particularly useful for imaging thick specimens. Two-photon microscopy utilizes two less-energetic photons to excite fluorophores, which reduces phototoxicity and photobleaching and increases imaging depth. Total internal reflection fluorescence (TIRF) microscopy is often used to observe events near the cell surface, using an evanescent wave that penetrates only a few hundred nanometers into the sample, thereby reducing background fluorescence and increasing contrast.
Live cell imaging provides high spatial and temporal resolution, allowing detailed analysis of cellular structures and it allows the study of dynamic cellular processes in real-time, providing a comprehensive understanding of cellular behavior and function. Since cells are observed in their natural state, the artifacts associated with cell fixation are eliminated. In addition, live-cell imaging systems facilitate longitudinal studies, allowing the same cell to be observed over long periods of time.
Live cell imaging technology has a wide range of applications in various fields. In cell biology, live cell imaging facilitates the study of cellular processes such as cell division, migration, adhesion, and intracellular trafficking. In neuroscience research, live cell imaging is used to observe neuronal growth, synapse formation, and neurotransmitter release. In developmental biology, live cell imaging can track cell lineages and differentiation during the development of an organism. In addition, live cell imaging is used in high-content screening assays to evaluate the effects of potential drugs on cellular processes and to advance the drug discovery process.
Amerigo Scientific offers live cell imaging systems with unique features to meet the diverse needs of cell biology, neuroscience, and other fields.
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