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The following guidelines will be helpful when choosing a filter set:
In simple terms for Flourescence microscopy you are trying to only allow a discreet portion of the light from the source to become incident on your sample and then to allow only that portion of the fluoresence emitted from the sample to reach the detector. Obviously a number of key points need to be considered as follows: |
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Spectral Characteristics of the Fluorophores |
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See our Fluorophore Reference Table to find the excitation and emission maxima of each fluorophore as well as the recommended filter sets. It is not always possible or desirable to use the peak maxima of the fluorophores but all the filter set designs have been optimised for the appropriate fluorphore.
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Sample Preparation |
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You need to know the relative strength expected (ie. bright or dim) from your labelled specimen. For routine visual applications where signal is not limiting, a Vivid Standard or Vivid Plus filter set is a good choice. For low-light level techniques, or when using a CCD camera, we recommend an Alpha Vivid set for maximum sensitivity. When labelling with multiple fluorophores you can use:
- a multi-band set for simultaneous visualisation of all dyes or proteins in the specimen.
- a hybrid pinkel set which makes use of single-dye exciters and a multi-band dichroic and emitter.
- E-mail Glen Spectra for choosing a single-dye set for your fluorophore.
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Applications |
Some techniques use specifically designed filter sets. Contact Glen Spectra for advice on filter sets for specific applications.
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Spectral Output From Your Excitation Source |
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Different light sources vary considerably in their output across the spectrum. As a result, some of our filter sets are better suited for mercury than xenon arc lamps, for example. See our light sources page for spectral output diagrams of the common lamps and lasers used in fluorescence microscopy.
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Spectral Response of Your Detector |
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Detectors have wavelength-specific sensitivities. CCD Cameras, for example, can detect IR light that the eye cannot. We provide filters that correct for such problems. See our Detector Response Curves for more information, or check with the manufacturer of your detector.
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Make and Model of Your Microscope |
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You will need to provide this information so that we can determine the correct filter sizes for your microscope. You should also specify if you would like your filters mounted in a filter holder. Glen Spectra offers filter sets to fit all microscopes. We sell holders for a number of microscopes and also mount filters in customer supplied holders. Please note that if you order a filter set and filter holder from us, we will mount the filters into the filter holder for free.
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