What is a fluorescence filter cube?
What is a fluorescence filter cube?
Filters provide a means of efficiently filtering the light that passes through the system, mostly in the form of fluorescent filter cubes. These cubes are specifically designed for one or several fluorophores of interest and are an essential component to high-quality fluorescent imaging.
How do fluorescence filter cubes work?
The function of the cube is to employ the excitation filter to tailor-make the excitation light reaching the fluorochrome; to ensure maximum reflection of the desired excitation light by the dichroic mirror; and finally to employ the barrier filter to pass the desired emission wavelengths yet block unwanted excitation …
What are the filters used in fluorescence microscope?
Most fluorescence instruments, including fluorescence microscopes, are based on optical filters. A typical system has three basic filters: an excitation filter (or exciter), a dichroic beamsplitter (or dichromatic mirror), and an emission filter (or barrier filter).
What is FITC filter set?
The Nikon dual excitation band DAPI-FITC filter set is designed for the simultaneous detection of the popular fluorophores DAPI and FITC (fluorescein isothiocyanate) when applied in combination, and incorporates an excitation filter with narrow bandpass windows in the violet (400-418 nanometers) and the blue (478-495 …
What is FITC fluorescence?
Fluorescein isothiocyanate (FITC) is a derivative of fluorescein used in wide-ranging applications including flow cytometry. FITC has excitation and emission spectrum peak wavelengths of approximately 495 nm and 519 nm, giving it a green color. Like most fluorochromes, it is prone to photobleaching.
How do I choose a fluorescence filter?
For optimal fluorescence detection when using a single dye, the excitation and emission filters should be centered on the dye’s absorption and emission peaks. To maximize the signal, one can choose excitation and emission filters with wide bandwidths.
What color is FITC?
green
FITC exhibits an excitation maximum at λ = 495 nm and emission maximum at approximately λ = 519 nm. The color of the compound is yellow while the emitted light is green.
What is an emission filter?
An emission filter serves the purpose of allowing the desirable fluorescence from the sample to reach the detector while blocking unwanted traces of excitation light. Like the excitation filter, this filter only allows a narrow band of wavelengths to pass through it, around the peak fluorophore emission wavelength.
Is FITC a 488?
Molecular Probes™ Alexa Fluor® 488 dye—with nearly identical spectral properties and quantum yield as fluorescein (FITC)—produces brighter, more photostable conjugates that are ideal for imaging and other applications requiring increased sensitivity and environmentally insensitive fluorescence detection.
What is the brightest fluorescent protein?
mNeonGreen
mNeonGreen: a yellow-green fluorescent protein mNeonGreen was reported as the brightest monomeric green or yellow fluorescent protein at the time. It is 1.5 to 3 times brighter than the most commonly used GFPs and YFPs.
What are fluorophores used for?
Fluorophores (or fluorochromes) are commonly used in conjugation with antibodies as detection reagents in applications such as flow cytometry. Fluorophores can absorb and emit light within a range of wavelengths, normally referred to as the absorbance (excitation) and emission spectra.
What are fluorescence filter cubes?
Fluorescence Filter Cubes Discover how variations in excitation and barrier filters allow certain wavelengths to pass. Usually, the lamp housing contains an infrared or heat filter to protect the fluorescence filters from heat deterioration.
What is the band pass of the FITC cube?
This cube has a 460-490 nm band-pass excitation filter, which will excite FITC. The barrier filter for this particular cube is NOT a long pass filter but a band pass of 515-550nm which will restrict the emission, reaching the eye or other detector, to the green wavelengths and will block red emission from the Rhodamine.
What is the excitation filter for the U-MNG cube?
A second cube, the U-MNG, has a band pass excitation filter 530-550 for green excitation of the Rhodamine conjugate. The barrier filter for the U-MNG cube is a long pass BA590 which will permit the red emission of the Rhodamine to reach the eye or other detector (e.g. film or video) and will block any green emission.
What are the different types of filter cubes?
The large product range includes longpass filter cubes for observation of single-stained samples and bandpass filter cubes suitable for detecting specific wavelengths from multiple-stained samples and un-mixing.