Andor's range of high performance imaging cameras are suited to a wide variety of microscopy applications, from extremely challenging single molecule tracking to routine wide-field fluorescence microscopy of living or fixed cells.
The market leading iXon range of EMCCDs provides ultimate sensitivity coupled with speed, the platform of choice for an array of light starved microscopy applications including single molecule tracking/interactions, photon counting, super-resolution microscopy, spinning disk confocal microscopy and TIRF.
Andor's innovative sCMOS cameras provide a great all round solution, redefining the concept of a 'workhorse' camera. Simultaneously offering extremely fast speeds, superb sensitivity, wide dynamic range, excellent resolution and a large field of view in one detector, sCMOS allows you to radically upgrade the performance of your fluorescence microscope. With the Andor sCMOS portfolio, we have your application covered, whether your samples are fixed or living, large or small, static or dynamic. sCMOS is also the perfect detector for Light Sheet microscopy.
Andor cameras are suitable for a variety of microscopy applications. Choose a slide from the list below to see how our ultra-sensitive detectors can benefit your critical research.
The ability to take light microscopy beyond the diffraction limit and resolve structures such as vesicles or molecules separated by less than 200 nm is the gift of super-resolution microscopy to cell biology. Among the main super-resolution techniques are PALM/STORM, STED microscopy, RESOLFT and SIM. In low light conditions image acquisition is a challenging task due to low-signal/high-noise nature of such experiments and the ability of the EMCCD camera to minimize the camera noise floor, even at fast readout speeds make them ideal. On the other hand with the emergence of brighter and more photo-stable fluorophores the high QE and extremely fast frame rates of Zyla 4.2 PLUS sCMOS make it an alternative choice for Super-resolution microscopy.
Fluorescence imaging of cells traveling in flow enables image based cell sorting and high-throughput microscopic characterization of extremely large numbers of cells. In addition, non-adherent cells can be imaged in conditions similar to the blood stream where hydrodynamic forces are known to play an important role in cell morphology and function. The temporal resolution of fluorescence imaging, however, is limited by a finite fluorescence emission rate. For a cell in fluid flow, this results in a trade-off between exposure time and motion-blur and sets a limit on the achievable signal to noise ratio of the captured image. High speed, high resolution sCMOS cameras are ideally suited, and the true global shutter functionality of the 5.5 megapixel sensor is essential to avoid readout distortion of the fast moving cells.
Case Study 1
Recent technical advances in optical detection and manipulation have made the detection of isolated, light emitting probe molecules a reality. Of particular importance to biology is the possibility of direct, real-time visualization of single biological macromolecules and their assemblies under native physiological conditions, offering great promise for enhancing our understanding of the behaviour, interactions, mechanisms and trafficking of individual biological macromolecules within the living cell. The raw sensitivity of the iXon range of EMCCD cameras ensures they are capable of detecting single photon events even at fast frame rates making them the ideal detector for the low light application of single molecule detection.
Over the past 10 years, a new fluorescence microscopy technique has come to the fore that is faster and less phototoxic than other microscopy techniques, making it ideal for studying living organisms and the biological processes that take place within them. Known as Light Sheet Microscopy (LSM) or Selective Plane Illumination Microscopy (SPIM), its key feature is that the illumination is done perpendicularly to the direction of observation. The field of view, resolution and speed of sCMOS make it an ideal detector for Light Sheet Microscopy.
Solutions for Light Sheet Microscopy:
Zyla 4.2 PLUS
TIRFM makes use of an optical effect that can be adapted to observe fluorescent events occurring at the interface between two optical media of different refractive indices. TIRFM is limited to the area within a few hundred nanometres of the glass/sample interface, where total internal reflection is occurring. TIRFM is an increasingly popular technique for visualizing, with high signal to background ratio, processes that occur in and around the membrane of living cells. Both EMCCD cameras and sCMOS (Zyla 4.2 PLUS) can be utilised for TIRF Microscopy. The sensitivity of the EMCCD is relevant when the fluorophores of choice are emitting a small number of photons, however when the photon emissions are sufficiently high, Zyla 4.2 PLUS achieves excellent resolution with a large field of view.
The ability to monitor changes in intracellular ion concentrations over time is vital for our understanding of signalling and functional pathways in cellular systems. Ion channels that span the outer cell membrane open or close in response to extracellular and intracellular signals, potentially altering how the cell behaves. These fluctuations can be visualized and quantified using ratiometric microscopy and special fluorescent dyes designed to bind specific ions. iXon EMCCD models ensure maximum sensitivity under fast frame rates, low dye concentrations and reduced illumination. Speed, resolution and field of view can be pushed to the max with the sCMOS cameras. Zyla 5.5 with Global Shutter exposure mode maintains temporal correlation across all areas of the sensor, e.g. when monitoring calcium sparks following electrical stimulation.
The application of an array of fluorochromes has made it possible to identify cells and sub-microscopic cellular components with a high degree of specificity amid non-fluorescing material. In fact, the fluorescence microscope is capable of revealing the presence of a single molecule. Through the use of multiple fluorophores, different probes can simultaneously identify several target molecules. Although the fluorescence microscope cannot provide spatial resolution below the diffraction limit of specific specimen features, the detection of fluorescing molecules below such limits is readily achieved. The large field of view (4.2/5.5 MP) and the excellent resolution (6.5 µm) of sCMOS make it an ideal detector for wide-field fluorescence microscopy.
Cell motility is required for many physiological processes during development, such as cell migration during gastrulation, axon guidance, tissue regeneration and embryological development. Of interest are whole cell movement, cell polarity, adhesion, membrane ruffles, protrusion of lamellipodia and filopodia, morphogenesis and also the involvement of the cytoskeleton. The sensitivity and speed of the iXon EMCCD cameras and sCMOS mean that both technologies can be successfully used for cell motility studies. In extremely low light conditions the raw sensitivity of EMCCD excels. However, if monitoring cell motility under transmitted light or DIC or if the fluorescence signal is sufficiently high sCMOS can also be used.
Case Study 1
Spinning disk confocal microscopy utilizes multiple pinholes to project a series of 1000 or more parallel excitation light beams onto the specimen in a multiplexed pattern that is subsequently detected after fluorescence emission passes through the same pinholes. The technique is highly useful for high speed imaging of living cells expressing fluorescent proteins. Photobleaching and phototoxicity are reduced with this technique, but a tighter light budget requires very sensitive camera systems for optimum signal detection. This is why EMCCDs are the camera of choice for this low light application. The EM gain functionality allows the user to preserve their living specimens and therefore image for longer time periods and the high sensitivity combined with the fast frame rates allow for dynamic imaging of extremely low light samples.
Fluorescence Recovery After Photobleaching (FRAP) denotes an optical technique capable of quantifying 2D lateral diffusion of a molecularly thin film containing fluorescently labelled probes or to examine single cells. This technique is very useful in biological studies of cell membrane diffusion and protein binding. Fluorescence Resonance Energy Transfer (FRET) is a microscopy technique used to measure the proximity of two fluorophores. Resonance energy transfer occurs only over very short distances, typically within 10 nm, and involves the direct transfer of excited state energy from the donor fluorophore to an acceptor fluorophore as an alternative to fluorescence emissive decay from the donor. Offering high resolution, high speed and high S/N, EMCCDs are the cameras of choice for FRAP and FRET imaging.
Live Cell Imaging now spans multiple modalities, including widefield (fluorescence, phase contrast, and differential interference contrast), laser scanning confocal, multiphoton, and spinning disk microscopy. It is used to provide critical insight into the fundamental nature of cellular and tissue function through the study of cellular dynamics. The EM gain capability of iXon EMCCD is a necessary requirement to ensure the living cell remains living for the duration of the experiment and special features such as 'Crop mode' acquisition provide fast frame rates for dynamic events. Alternatively the speed and field of view of sCMOS make it ideal for live cell imaging. Zyla 5.5 with Global Shutter exposure mode provides a capability that is both 'photon-efficient' and easy to synchronize to, especially critical for 3D / 4D microscopy.
Bioluminescence refers to the ability of living things to produce light and represents a profoundly useful area of biophotonic research. A well-known bioluminescence is that induced by the luciferase enzyme. For example, firefly luciferase emits light in the presence of its substrate luciferin & adenosine triphosphate (ATP) and is widely used for measuring ATP concentrations. Low read noise, low dark current and a high QE are key parameters for a bioluminescence detector. Deep cooled CCD cameras are suited to ultrasensitive imaging of bio/chemiluminescence. They offer ultra-low dark noise due to exceptional TE Cooling and are ideal if longer exposures are required for weaker bioluminescent signals.
Case Study 1
Super Resolution Microscopy
Ion Signalling Microscopy
Spinning Disk Confocal Microscopy
Live Cell Imaging
Wide-field Fluorescence Microscopy
Single Molecule Detection
FRAP / FRET
Light Sheet Microscopy
Andor offers the leading portfolio of high performance low light imaging camera technologies, born of consistent innovation, unparalleled quality and reliability, and superlative customer service.
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