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Introduction to Fluorescence Filters - Optical fluorescence occurs when a molecule absorbs light at wavelengths within its absorption band, and then nearly instantaneously emits light at longer wavelengths within its emission band.
Multiband Filter Set Terminology - To satisfy the ever-increasing demand for high-speed imaging, especially for live-cell real-time analysis using fluorescent protein labels, there is a need for an alternative to the single-band filter cube approach without sacrificing image fidelity.
Fluorescent Proteins Theory Applications and Best Practices - First identified in 1962 in sea creatures, fluorescent proteins have proved versatile and extremely useful, as demonstrated by applications they either enable or significantly benefit.
Choosing Dichroic Beamsplitters with Flatness/ RWE Appropriate to the Microscopy Method - Wavefront distortion can degrade image quality by reducing contrast or compromising resolution. In several microscopy applications, reducing wavefront distortion is critical to achieving the microscopy method. Specifying and selecting optical filters that minimize wavefront aberration is important to maximize or enable optical system performance. This technical note elucidates how to select optical filters for high performance microscopy, and provides guidance on choosing Semrock catalog filters for wavefront distortion performance required for applications.
Super-Resolution Dichroic Beamsplitters - Semrock sets a new standard for super-resolution microscopy with λ/5 P-V RWE on our new 3 mm thick dichroics and improved 1λ P-V per RWE on our improved 1 mm dichroics. These industry leading optics improve the performance of laser based confocal and TIRF illumination systems, and are ideal for reflection of imaging beams in conventional structured-illumination techniques as well as patterned illumination systems for localized photo-activation.
Image-splitting Dichroic Beamsplitter - Semrock offers the industry standard enabling simultaneous multi-color imaging applications. The spectral edges of these filters are optimized for imaging of popular fluorophore-pairs providing maximum signal throughput, while maintaining minimal wavefront distortion in reflection and transmission thereby maximizing contrast and resolution of the overall imaging system.
Super-resolution Microscopy Cubes – Super-resolution Microscopy Cubes set the new standard for laser based microscopes. These cubes are optimized for mounting 1λ RWE 1mm thick super-resolution laser dichroic beamsplitters. Maximize SNR and minimize artifacts in TIRF, Confocal, PALM, STORM, SIM, and other super-resolution techniques.
Flatness of Dichroic Beamsplitters - Glass substrate is not always perfectly flat, especially after it is coated, since the intrinsic stress of hard glass coatings can cause slight bending of the substrate. This bending can cause focal plane shift and image distortion when imaging reflected light.
Practical Flatness - BrightLine® laser dichroic beamsplitters set a new standard for super-resolution microscopy with λ/5 P-V Reflected Wavefront Error (RWE) performance. We’ll explain how dichroic beamsplitter flatness affects an optical wavefront in reflection, and how to calculate the practical impact of flatness specifications on your system.
VersaChrome Edge™ tunable filters
unlock virtually unlimited spectral flexibility for fluorescence microscopy and hyperspectral imaging, as well as for spectroscopy applications allowing users to create a bandpass filter as narrow as sub 5nm FWHM or as wide as 12% of the center wavelength throughout the visible and near-infrared wavelength ranges.
Tunable Bandpass Filters - Semrock has now developed a revolutionary new optical filter technology: thin-film filters that are tunable over a wide range of wavelengths by adjusting the angle of incidence with essentially no change in spectral performance.
Spectral Imaging with VersaChrome Filters - Using tunable bandpass filters in spectral imaging systems in a range of applications.
Fluorescence Resonance Energy Transfer - Fluorescence Resonance Energy Transfer (FRET) is a powerful technique for characterizing distance-dependent interactions on a molecular scale. It is one of the few tools available that is able to measure intermolecular and intramolecular distance interactions both in-vivo and in-vitro.
Optical Filter Configurations for FRET - Explanations of the classic and most popular approaches to the FRET method of imaging.
Fluorescence Imaging with Quantum Dot Nanocrystals - Quantum dot nanocrystals are fluorophores in that they absorb photons of light and then re-emit longer-wavelength photons, however, there are some important differences between quantum dots and traditional fluorophores.
Ultraviolet Fluorescence Applications - Many biological molecules of interest naturally fluoresce when excited by shorter- wavelength UV light. Because the fluorescence is intrinsic, samples can be observed without the added chemistry and limitations associated with "indirect" labeling by extrinsic fluorophores.
Using Fura-2 to Track Ca2+ - The fluorophore Fura-2 has an absorption spectrum that varies markedly depending on the concentration of calcium (Ca2+) that is present near the fluorophore molecule.
Using Fura-2 to Track Calcium Using VersaChrome Filters - Ratiometric imaging with Fura-2 can be more carefully optimized to your specific experimental conditions when using tunable bandpass filters.
BrightLine® Multiphoton LaserMUX™ Beam Combiners - Multiphoton LaserMUX™ beam combiners enable deeper tissue imaging and improved contrast in multi-color and multi-modal fluorescence microscopy.