Capillary electrophoresis fluorescent detection system
View Patent ↗The invention is a high sensitivity and high throughput capillary multiwavelength fluorescence detection system. The fluorescent detection system is configured to illuminate a relatively large volume of a single capillary or a plurality of capillaries, with a pixelated detection system capable of imaging an area of each capillary that differentiates the capillary walls, the space between the capillaries, and the internal liquid volume within the capillary. Only the desired pixels or image area are used for processing and generating an output signal.
1. A fluorescence detection system, comprising:
a sample vessel comprised of capillaries;
an LED light source to emit light to excite a fluorescently labeled sample contained in said capillaries, the system configured to direct light on the sample vessel to illuminate the sample;
a CCD fluorescence detector positioned to detect the fluorescent emissions of the sample, said fluorescence detector capable of producing an image, having sufficient resolution to detect three distinct regions of the image, including a wall of each capillary, an internal volume of each capillary, and a space between the capillaries, each region being defined by at least one pixel, and
a processor capable of providing an integrated signal for selecting which pixels to process for an integration of light intensity, with pixels from the walls of the capillary and pixels from the space between the capillaries specifically excluded from the integrated signal; and
integrating of the light intensity of said pixels to provide a fluorescent signal.
2. The fluorescent detection system of claim 1 , wherein said sample vessel of said system contains at least two separate windows, and the detection system includes at least two light sources, and the fluorescent light from each of the said two separate windows is imaged onto at least two separate areas of the CCD.
3. The fluorescence detection system of claim 1 , wherein the system is configured to direct light on more than one sample vessel.
4. The system of claim 1 configured so that the width of at least one pixel defines the volume of each capillary.
5. The system of claim 1 that uses multiple detection windows.
6. The CCD fluorescence detection system of claim 1 , with said system being configured to detect fluorescent emissions at more than one wavelength.
7. A method of detecting a fluorescence emission in a sample the method comprising:
introducing a fluorescently labeled sample into a sample vessel;
directing a light from an LED light source to excite the fluorescently labeled sample, the light directed on the sample vessel to illuminate the sample vessel; and
detecting a fluorescent emission from the fluorescently labeled sample with a CCD fluorescence detector capable of producing an image having sufficient resolution to detect three distinct regions of the image;
said three regions including a wall of each capillary, an internal volume of each capillary, and a space between the capillaries, each region being defined by at least one pixel, and
using a processor capable of providing an integrated signal for selecting which pixels to process for an integration of light intensity, with pixels from the walls of the capillary and pixels from the space between the capillaries specifically excluded from the integrated signal.