This dye and the related dye, Di-4-ANEPPS, are most effectively used as ratiometric indicators of membrane potential. Increases in membrane potential are indicated by a decrease in fluorescence intensity with approx. 440nm excitation and an increase in fluorescence intensity with approx. 530nm excitation. Use of this 71006 filter set requires an external filter wheel to house the 2 excitation filters and to facilitate rapid switching of filters for ratiometric measurements.
This dye has mainly been used as a retrograde neuronal tracer, although it can also distinguish between DNA and RNA binding according to its fluorescence emission. With two excitation peaks at approx. 330nm and 390nm, and two emission peaks at approx. 450nm and 600nm, our DAPI longpass filter sets 49025 and 19012 are appropriate for detection of the fluorescence emission associated with the both peaks, which together appear "gold". The 49025 set is also useful for distinguishing between FluoroGold bound to DNA or RNA.
The spectral properties of ICG are strongly dependent on its concentration and the solute it is bound to, so care must be taken by workers to characterize their particular model system. This filter set 49030 is appropriate for most applications involving protein-bound ICG in various physiological media. ICG becomes concentrated in lipid environments, greatly shifting its spectral properties. 49030 may be used with LEDs with CWL 755-795nm.
JC-1 exists as a monomer at low concentrations and/or low membrane potentials and displays fluorescence ex./em. maxima similar to that of YFP. At higher concentrations and/or higher membrane potentials, JC-1 forms aggregates with a red-shifted emission maxima of 590nm and a broadened excitation spectra. The 49012 longpass filter set will allow detection of JC-1 monomer and aggregate simultaneously. 49012 may be used with LEDs with CWL 465-495nm; 19002 may be used with 470-490nm CWL LEDs.
JC-1 exists as a monomer at low concentrations and/or low membrane potentials and displays fluorescence ex./em. maxima similar to that of YFP. At higher concentrations and/or higher membrane potentials, JC-1 forms aggregates with a red-shifted emission maxima of 590nm and a broadened excitation spectra. Use of this 71019 filter set requires an external filter wheel to house the 2 emission filters to facilitate rapid switching of filters for ratiometric measurements.
Calcofluor White is a commonly used stain to detect fungi and yeast and other microorganisms. It is a non-specific stain which preferentially binds to chitin and cellulose in cell walls and as such is also used to visualize plant cells. Calcofluor White has extremely broad excitation/emission spectra, and as a result almost any filter set with UV or violet/blue excitation wavelengths will generate fluorescence emission which may be detected across a very broad range. Some samples will fluoresce strongly in the green wavelengths, others in the blue, and others yellow-orange. Because of this, filter selection is subjective, partly depending on the experience of the microscopist. Our filter sets 19012, 19000, and 19011 may be used successfully depending on the sample and the observer's preference. 19000 and 19012 may be used with LEDs with CWL 365-380nm; 19011 may be used with 395-410nm CWL LEDs.
EGFP spectra used here; similar to Emerald GFP, TagGFP, Azami Green, mWasabi, etc.49002 may be used with LEDs with CWL 455-485nm; 49020 may be used with 475-485nm CWL LEDs; 39002 may be used with 470-490nm CWL LEDs.
If workers must detect fluorescence from both RNA and DNA-bound Acridine Orange a longpass set such as 49012 is recommnded. 49011, 39002 may be used with LEDs with CWL 465-495nm; 49002 may be used with 455-485nm CWL LEDs.
For imaging applications using a monochrome camera, this longpass filter set will not distinguish FM 4-64 from TRITC or similar orange/red fluorochromes. To be able to distinguish between FM 4-64 and TRITC or Texas Red-like fluorochromes, contact us for a custom filter set. 19010 may be used with LEDs with CWL 470-490nm; 19006 may be used with 545-575nm CWL LEDs.