Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions

BMC Biotechnology

Table 1 Relative brightness of DAPI and QDs using various fluorescence filter set configurations

Filter Set Configuration	A	B	C	Modified C
Filters
Excitation	DAPI	DAPI	DAPI	DAPI
Dichroic	Triple	Triple	DAPI	DAPI
Emission	Triple	DAPI	DAPI	QDot 655
Light Output
DAPI	135	135	269	0
QDot 585	6,300	252	42	490
QDot 655	36,637	983	1,747	27,846
Ratio QDot 655: DAPI	271	7.3	6.5

Light output values take the emission lines of the mercury arc lamp, filter sets, and dye absorbance and emission spectra into account to determine the light output of particular fluorophores given a particular optical configuration. Note that a "Triple" filter and dichroic refers to a standard multichroic or emission filter with multiple bandpasses for DAPI/FITC/TRITC, typical on a cytogenetics workstation. Light output values were obtained using "Fluorescent Spectra: An Interactive Exploratory Database", a web-software tool developed by C Boswell and G McNamara http://www.mcb.arizona.edu/ipc/fret/.

ISSN: 1472-6750