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Fluorescence Filters

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Flourescence filters are at the heart of every flourescene optical device, and high quality filter sets are essential for high definition imaging and fluorescence microscopy. At Shanghai Optics we produce fluorescence filters for a wide range of applications in the form of filter cubes and filter wheels.

Flourescence Microscopy

Fluorescence microscopes are designed to detect the flourophores in a sample; either naturally occurring flourophores or those used as dyes and labels. These flourophores absorb energy in specific wavelengths (excitation range), and emit it in a different wavelength (emission range). This allows us to identify fluorophores by absorption and emission.

More precisely, as each flurorophore has a unique excitation spectrum and a unique emission spectrum, each peaking at a specific wavelength, they can be distinguished by either their excitation spectrum, by their emission signal, or by their Stokes shift, which is the distance between peak excitation and peak emission. This is the basic principle behind all fluorescence microscopy systems, whether it be a simple system such as a epifluorescent microscope or a more complex setup such as those used in multiphoton systems.

Flourescence Filter Sets

Each fluorescence filter cube features three distinct optical filters: an excitation filter a dichroic mirror and an emission filter.

The excitation filter, otherwise called an exiter, is a bandpass filter designed to pass only the wavelengths of light which are absorbed by the fluorophore. By doing so, it minimizes excitation that might be caused by the light source and other sources of fluorescence and also blocks excitation wavelength light in the flurorecence emission band. The minimum transmission of this filter is what will determine the brightness or brilliance of the final image. In general, an excitation filter should have a minimum of 40% transmission; transmission of greater than 85 percent will give a bright, well-lit image. Optical density determines the background darkness, and should be at minimum 3.0 but preferably closer to 6.0. The excitation filter should be chosen so that the center wavelength (CWL) is close to the peak excitation of the flourophore, and the bandwith is completely within the flurophore excitation range.

The dichroic mirror is a specialized edge filter sometimes called a dichromatic beamsplitter. When used at a 45 angle of incidence, it is able to reflect light in the wavelengths of the excitation band and transmit light from the emission band to the emitter and on to the detector.

The third component of the fluorescence filter is the emitter. This is also an bandpass filter, and passes exclusively light emitted by the flurorophore. Excitation light and all other light outside, both UV an IR, are blocked out effectively by the emitter filter.

Our standard high performance fluorescence filter cubes are carefully designed to pass the appropriate wavelengths for high-brightness imaging while blocking irrelevant light that might decrease contrast. Since both system autoflourescence and excitation light is blocked, our filters are able to provide a high signal to noise ratio and the dark background needed for precise imaging.

When a bright image is more crucial than high contrast, a longpass edge filter may be used as an emission filter in place of the bandpass filter. Longer wavelengths of light will then be transmitted, producing a well-lighted brighter imager.

When a multiband excitation filter is combined with a multiband emission filter and a polychroic rather than dichroic beamsplitter image with all fluorescent tags will be formed. This type of filter set is referred to as full-multiband and can be used with multiple fluorophores.

Choosing a Flourescence Filter

At Shanghai Optics we produce a wide range of fluorescence sets that are designed with the excitation and emission filters suitable for each commonly used fluorophore. For specialized applications, such as those using multiple flurophores, alternate dichroic filters or laser sources, we are able to produce custom filter sets upon request. Please contact us to discuss the specific requirements of your application and to get a free consultation and a price quote.

Key Features

  • High passband transmission (T>90%)
  • Excellent blocking (OD>6.0)
  • Hard coatings and no adhesives for long filter life
  • All dielectric coated with IBS technology

Application

  • Fluorescence microscope
  • Co-localization fluorescence measurements
  • Fluorescence In Situs Hybridization (FISH)
  • Comparative Genomic Hybridization (CGH)
  • Gel and spot Imaging comparisons

Flourescence Filter Factory Standards

Diameter: 18.0, 20.0, 22.0, 25.0mm Diameter Tolerance: +0/-0.1mm
Passband Transmission: T>90% Clear Aperture: >80%
Surface Quality: 60-40 Angle of Incidence: 0° +/- 5°
Beam Deviation: <11 arc sec. Transmitted Wavefront: 1/4λRMS@633nm
Operating Temperature: -45°C~85°C Environmental Durability: MIL-STD-810F
Temperature Dependence: <5ppm/°C Blocking OD>6.0(typical): UV-700nm, λc<500nm UV-925nm, λc>500nm
Physical Durability: MIL-C-48497A Substrate: Low Fluorescence BK7 or Equivalent

 

Central Wavelength(nm) Bandwidth Transmission(%) Thickness(mm) Typical Application
438 24 90% 5.0 CEP Excitation
447 60 90% 3.5 DAPI Emission
472 30 90% 5.0 GFP Excitation
482 35 90% 5.0 FITC Excitation
483 32 90% 3.5 CFP Emission
500 24 90% 5.0 YFP Excitation
510 84 90% 3.5 FURA2 Emission
520 35 90% 3.5 GFP Emission
531 40 90% 5.0 CY3 Excitation
536 40 90% 3.5 FITC Emission
542 27 90% 3.5 YFP Emission
543 22 90% 5.0 TRITC Excitation
562 40 90% 5.0 Texas Red Excitation
593 40 90% 3.5 TRITC/CY3 Excitation
624 40 90% 3.5 Texas Red Emission
628 40 90% 5.0 SY5 Excitation
692 40 90% 3.5 SY5 Emission

 

fluoscence filter
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