Leica Upright Light Microscopes These powerful imaging systems feature constant color, natural light illumination, superior optics, and configurable options to provide high contrast, brilliant images for your biological research
Microscopy is the foundation of basic biology research. The ability to see biology supports the needs of the scientific community with the visualization, measurement and analysis of cells and particles. It is one of the most ubiquitous piece of equipment found in the life science lab. Specifications can range from simple upright widefield microscopes to powerful confocal fluorescence or super-resolution microscopes.
Microscopy is used in the daily work of most cell biology labs in fields such as oncology, virology and neuroscience. The basis of human health and its associated diseases can be understood from a cellular level with remarkable dynamic, molecular detail. The study of tissues and live specimens on a macroscopic scale gives us biological insight, as well as analyzing individual cells, cellular organelles and macromolecules.
In the life sciences, your popular choices are between upright widefield microscopes, inverted microscopes for cell culture applications, and stereo microscopes for macro-viewing applications.
Fluorescence is the most commonly used way of tagging and identifying single molecule species. The technique uses the luminescence phenomena found in nature to stain entire cells or biomolecules with fluorophores, allowing for the fluorescence microscope to detect the signal, generating both spatial and intensity data. This data is representative of the localization and the amount of the molecules inside a cell. Colocalization and interaction studies can be performed.
Live cell imaging is a popular technique to study live cells and investigate biological processes in real-time. Using time-lapse microscopy and careful preparation of cell environments, it is possible to observe cell-to-cell interactions, the behavior of single cells, and changes within the cell. Any imaging technique can be used to address live cells, with the most popular being phase contract microscopy, fluorescence and confocal microscopy, and light sheet microscopy. Live cell imaging can be performed inside a CO2 incubator with microscopes designed to fit and operate inside a typical incubation chamber and can be performed outside with special stage-top incubators.
Live cell imaging is suited for the study of:
The primary hurdles in live cell imaging is to balance the photo-capture capabilities and therefore image quality with preserving cell health. Suitable technologies and consumables are available to ensure optimal environments that live cell imaging experiments perform well.
Organoids and 3D cell cultures are an exciting development that combines bioengineering with life sciences. Recent advancements have created stable cell cultures in three dimensions, such as organoids, spheroids, or organ-on-a-chip models. 3D cell cultures are grown or printed to mimic the in vivo environment of cells inside the body. Animal cells are embedded in the extracellular matrix (ECM), which is composed of proteoglycans and fibrous proteins. This complex, dynamic, and tissue-specific 3D structure provides physical scaffolding for the cells and initiates cues that influence cell differentiation and behavior.
The majority of cells in living tissue grow, communicate, move, and receive nutrients and oxygen in a physically 3D environment. Therefore, cells behave differently inside a 3D gel matrix compared to a 2D environment. In many cases, a 3D environment reflects the in vivo situation more accurately. This should be considered when analyzing cell behavior, differentiation, response to drug treatment, and gene and protein expression.
Super-resolution microscopy enables the visualization of the smallest structures in living cells that cannot be resolved using standard widefield or confocal fluorescence microscopy. This technique provides a spatial 3D resolution that is well below the diffraction limit. It creates new views on the structural organization of cells and the dynamics of biomolecular assemblies, that are closer to a near-molecular resolution.
When using widefield and confocal fluorescence microscopy, the diffraction barrier limits the maximal resolution to about 200 nm. Super-resolution microscopy breaks the diffraction barrier, enabling “nanoscopy” with a substantially improved optical resolution of down to 5–20nm. This method uses the physical or chemical properties of adjacent fluorophores to resolve them from each other. For example, while one fluorophore’s state is “on”, the neighboring fluorophore’s state is “off”, which enables their differentiation.
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