Improved fluorophore separation with IMS confocal microscopy.
Some of the most extensively used fluorophores for multicolour fluorescence labelling of tissue, such as Lucifer yellow, fluorescein, Cyanine-3.18 and rhodamine, all have the disadvantage of being difficult to separate spectrally due to their broad and overlapping emission spectra. With the intensity-modulated multiple-beam scanning (IMS) technique, cross-talk between labels can be reduced to a fraction of that seen using conventional excitation and detection technique. Thus, cross-talk to the 'red' channel decreased with about one order of magnitude for the 'green' fluorophores Lucifer yellow and FITC. For the tested 'red' fluorophores, the amount of cross-talk to the green channel was reduced by 60-90%.
Carlsson
K
K
Physics IV, Royal Institute of Technology, Stockholm, Sweden.
Ulfhake
B
B
D000818:Animals; D019793:Fluorescein; D005452:Fluoresceins; D007150:Immunohistochemistry; D007546:Isoquinolines; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology*; D051381:Rats; D011919:Rats, Inbred Strains; D013116:Spinal Cord / Q000166:cytology*
D005452:Fluoresceins; D007546:Isoquinolines; C017475:lucifer yellow; D019793:Fluorescein