Live-cell imaging is a microscopy technique that allows in vivo imaging of cells, instantly and over a period of time. There are different types of microscopy compatible with live-cell imaging, which include both conventional contrast techniques, like differential interference contrast (DIC) or phase contrast, and fluorescence-based techniques.
The most commonly used are those capable of performing live-cell fluorescence microscopy, which allow researchers to study and follow the localization of fluorescence proteins in cells and in different tissues. Amongst these, scientists can use epi-fluorescence or widefield fluorescence, laser scanning or spinning disc confocal, lattice light-sheet and various super-resolution microscopy techniques, such as SIM or PALM.
A microscope adapted for live-cell imaging will include whole body heating or a heating device connected to the stage to keep cells at physiological temperatures and, when necessary, a CO2 incubator to support imaging over extended periods of time.
Live-cell imaging can provide a large amount of information and detail about a molecule of interest, cellular organelle or cell type within a tissue. Imaging of fixed samples, like those labeled with immunofluorescence staining, often only provide a snapshot of a specific cellular process or the localization of a molecule. This can limit the interpretation of information, particularly when studying highly dynamic processes.
With live-cell microscopy techniques, scientists can obtain highly valuable information of such biological processes, including:
> Molecular localization and movement with time resolution
> Dynamic complex assembly and interaction between molecules
> Organization of components within cells
> Vesicle or viral particle behavior
> Cellular responses to environmental cues
The Nanoimager allows whole body heating and adjustable temperature settings, including 37℃ and up to 42℃, to support live-cell imaging of different cell types in various organisms. Its compact design, unrivaled stability and smart alignment design, makes it a robust microscope that can be used in any laboratory environment.
The Nanoimager offers powerful imaging of single molecules with enhanced resolution of up to 20nm by supporting different super-resolution techniques, including dSTORM, PALM, single particle tracking, smFRET, confocal and SIM microscopy. For live imaging, SIM is easy to use and offers high speed imaging, while smFRET provides quantitative information on dynamic protein-protein interactions. PALM microscopy offers even better resolution and can be combined with HILO or TIRF microscopy to improve the signal to noise ratio of thinner samples.
The Nanoimager's NimOS software includes a single-particle tracking feature, which provides tracking analysis, particle size and number measurements, and automated detection over time. Our microscope can image two fluorophores simultaneously (with four laser colors) on a single sample, allowing tracks registered in one channel to be assigned to cellular markers in the second channel.
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