Fluorometer with low heat-generating light source
This invention concerns a fluorometer preferably combined with a thermal cycler useful in biochemical protocols such as polymerase chain reaction (PCR) and DNA melting curve analysis. The present fluorometer features a low heat-generating light source such as a light emitting diode (LED), having a one-to-one correspondence to each of a plurality of sample containers, such as capped PCR tubes in a standard titer tray. The fluorometer of the present invention further comprises an optical path between each LED and its correspondingly positioned container, and another optical path between each fluorescing sample within the positioned container and an optical signal sensing means. The instrument can be computer controlled.
1. A fluorometer comprising:
a plurality of light sources, wherein each of said light sources generates heat at a level below which active cooling is required;
a plurality of containers for containing a potentially fluorescing sample;
a first optical path means;
a second optical path means; and
a means for sensing optical signals,
wherein there is an equal number of said light sources and said containers, each of said containers corresponds to one of said light sources, and said fluorometer does not comprise a positioning mechanism for repeatedly presenting said containers to said light sources during operation of said fluorometer; and
wherein said first optical path means is located between said plurality of light sources and said plurality of containers, said first optical path means is straight or relatively straight, said first optical path means does not comprise a fiber optic line, said second optical path means is located between said plurality of containers and said means for sensing optical signals, and said means for sensing optical signals is in optical communication with said containers.
2. The fluorometer of claim 1 , wherein said first optical path means comprises a lens.
3. The fluorometer of claim 1 , wherein said second optical path means comprises a lens.
4. The fluorometer of claim 1 , wherein said first optical path means comprises a lens and said second optical path means comprises a lens.
5. The fluorometer of claim 1 , wherein said first optical path means comprises a plurality of lenses, wherein each of said plurality of lenses corresponds to one of each of said light sources.
6. The fluorometer of claim 1 , wherein said second optical path means comprises a plurality of lenses, wherein each of said plurality of lenses corresponds to one of each of said containers.
7. The fluorometer of claim 1 , wherein said first optical path means comprises a first plurality of lenses and said second optical path means comprises a second plurality of lenses, wherein each of said first plurality of lenses corresponds to one of each of said light sources and each of said second plurality of lenses corresponds to one of each of said containers.
8. The fluorometer of claim 1 , wherein said first optical path means is located in proximity to said light sources relative to said second optical path means and said plurality of containers, and said second optical path means is located in proximity to said containers relative to said first optical path means and said plurality of light sources.
9. The fluorometer of claim 1 , wherein said light sources are light emitting diodes.
10. The fluorometer of claim 9 , wherein said light emitting diodes are blue.
11. The fluorometer of claim 1 , further comprising a programmable controller for controlling at least one function of the fluorometer.
12. The fluorometer of claim 1 , wherein said containers comprise sample tubes.
13. The fluorometer of claim 12 , wherein said sample tubes are held in position by a sample tube holder or a multiplate.
14. The fluorometer of claim 13 , wherein said sample tubes form an array, and said light sources, first optical path means, and second optical path means are arranged in corresponding arrays.
15. The fluorometer of claim 14 , wherein said array is an 8 by 12 sample array.
16. The fluorometer of claim 1 , wherein said means for sensing optical signals is selected from the group consisting of a photomultiplier tube, a charge coupled device type camera, a vacuum photodiode, and an avalanche photodiode.
17. The fluorometer of claim 1 , further comprising at least one reflector positioned to guide light emitted by a fluorescing sample from at least one container to said means for sensing optical signals.
18. The fluorometer of claim 1 , wherein at least one of said plurality of containers contains a potentially fluorescing sample.
19. A method for analyzing a sample, comprising the steps of:
(a) providing a sample in at least one of the plurality of containers in the fluorometer of claim 1 ;
(b) generating light from at least one of the plurality of low heat generating light sources;
(c) guiding the generated light from said at least one light source through its corresponding first optical path means to its said corresponding container in which said sample is provided; and
(d) detecting for a fluorescing signal from said sample with said means for sensing optical signals.
20. An apparatus comprising:
(a) the fluorometer of claim 1 ; and
(b) a thermal cycler.