Fluorescence generation device
The present invention relates to a kind of light generating device, more particularly to a kind of fluorescence generation device for fluorescence generation device technical field.When background technology carries out science of heredity, the gene studies such as molecular biology or the detection of animals and plants epidemic disease, need first with the means of amplification of nucleic acid (；Such as polymerase chain reaction, Polymerase Chain Reaction, PCR), a small amount of sample of nucleic acid is replicated into amplification to the amount that can be detected in a short time.Above-mentioned nucleic acid amplification product can further make target nucleic acid fragment with linking with fluorescence, radiogen or in the nucleic acid probe (probe) of chromogenic enzyme via hybridization (Hybridization), produce fluorescence, irradiation image or color reaction.Use fluorescein stain as marker on many biochips at present, preferable analysis result can be provided by being primarily due to fluorescein stain, if compared with traditional colour generation agent method, fluorescein stain can provide high about 1000 times to 500,000 times of sensitivity.Above-mentioned fluorescence reaction is observed, detected and analyzed to the devices such as fluorescence microscopy (Fluorescence Microscopy) fluorescence analyser, flow cytometer, camera (Camera) or image capture unit to capture after fluoroscopic image.These technologies are directed to the selection of excitation source, because specific fluorescein stain needs to use the light of particular range of wavelengths to come as excitation source, when the light irradiation of an appropriate wavelength has the molecule of photoluminescent property, molecule can light absorbing energy and be excited to higher-energy state, and in very short time (10_8〜10_4Second) in return back to low-energy state, excess energy is disengaged in the form of giving out light simultaneously, therefore matching excitation source must be selected according to the characteristic of fluorescein stain, optimal stain stimulation effect can just be obtained, and (selection of Excitation^ excitation sources includes sending ultraviolet light or laser light etc. with general light source, but ultraviolet light is easily scattered, and transmission and penetrate and be difficult, therefore, detecting instrument must use special optical component with highly ultraviolet luminous sensitivity, so that its cost is high, economic benefit is not met.And if using laser light as excitation source, it has, and wavelength is single, penetrability is high and is easily detected, but laser exciting light must coordinate filter and it is spectroscopical use, its volume is occupied greatly, and make it that the erection of its instrument is difficult.Therefore, as TaiWan, China patent discloses " excitation source to excite fluorescent signal " of No. 201018728, it discloses a kind of by the use of adjustable luminous intensity and the light-emitting diode (LED) module of photochromic combination as excitation source, and the mode through photochromic combination adjustment obtains and can excite the excitation wavelength of fluorescein stain.But fluorescein stain only can just obtain preferably launching efficiency under the excitation source of specific wavelength, and the wave-length coverage of the light source of light-emitting diode (LED) module easily excited with fluorescein stain after fluorescent wavelength ranges have the overlapping of part, and the intensity for causing to cannot be distinguished from light signal when measuring is the light source of fluorescence after exciting or light-emitting diode (LED) module, the problem of accuracy is not good is caused.
The content of the invention main object of the present invention, is the excitation source for providing particular range of wavelengths, the generation efficiency of fluorescence is excited with effective lifting.It is another object of the present invention to the wave-length coverage for solving existing excitation source is easily overlapping with the wave-length coverage of fluorescence light source, and the problem of cause fluorescence excitation amount detection difficult.To reach above-mentioned purpose, the present invention provides a kind of fluorescence generation device, including sends the blue light diode of light beam, optical filtering part and fluorescein stain.The light that the optical filtering part is arranged at the blue light diode projects direction, to receive the light beam and change output filtering light beam, the wavelength of the filtered beam is between 465nm between 505nm, and the fluorescein stain is arranged at the optical filtering part with respect to the side of the blue light diode to receive the filtered beam and inspire fluorescence.As shown in the above description, compared with prior art, the present invention has following features：First, the wave-length coverage of optical filtering part control filtered beam and excites the fluorescein stain to obtain fluorescence in 465nm between 505nm by high efficiency and in the way of being precisely controlled wavelength.2nd, the wave-length coverage of filtered beam is set, the wave-length coverage of filtered beam can be avoided overlapping with fluorescent wavelength ranges or partly overlapped, the error problem on measuring is caused.Fig. 1 is illustrated, is the configuration block schematic diagram of the preferred embodiment of the present invention；Fig. 2, is the dimensional structure diagram of the preferred embodiment of the present invention；And Fig. 3, it is the spectral wavelength schematic diagram of the preferred embodiment of the present invention.Embodiment detailed description for the present invention and technology contents, it is as follows in conjunction with illustrating：As shown in Figures 1 and 2, the invention provides a kind of fluorescence generation device, including the blue light diode 10, optical filtering part 20 and fluorescein stain 30 of light beam 11 are sent.The light that optical filtering part 20 is arranged at blue light diode 10 projects direction to receive light beam 11 and change output filtering light beam 21, the wavelength of filtered beam 21 is between 465nm between 505nm, the fluorescein stain 30 is arranged in test tube 32, and make the test tube 32 be placed in the optical filtering part 20 with respect to the side of the blue light diode 10 to receive the filtered beam 21 and inspire fluorescence 31, detection module 40 carries out the detection of spectrum, in the present embodiment, blue light diode 10, optical filtering part 20 and detection module 40 are connected with computer 50
Computer 50 controls the luminous intensity and switch, the filter range of optical filtering part 20 and the information for receiving the detection module 40 of blue light diode 10.In addition, the present invention is additionally provided with the progress that heating member 60 is reacted with sharp PCR.In general, as shown in Figure 3, the display that light wave is longer than in frequency spectrum is that have peak value within the specific limits and gradually decay from the peak value toward both sides, and the numerical value of wavelength only refers to the numerical value corresponding to the peak value, and not representing light has value in single wavelength and impulse form is presented.For example, the wavelength alleged by the present invention is 488nm, refers to the peak value of its light wave at 488nm, and still has value in 488nm front and rear wave band.Therefore, if the optical source wavelength of blue light diode 10 is not single enough, the overlapping wavelengths of fluorescence 31 after easily causing the part wave-length coverage for the light beam 11 that blue light diode 10 is sent and exciting, so that the detection module 40 receives the light energy of the light beam 11, the problem of causing detection error in detection in the lump.The present invention is filtered and obtained the excitation wavelength of filtered beam 21 and correspondence fluorescein stain 30 through optical filtering part 20 to blue light diode 10, and the fluorescein stain 30 can be 6- fluorescein phosphoramidates（6-FAM), 5- fluoresceins phosphoramidate（5-FAM), Oregon green -488 (Oregon Green-488), A Laisha -488 (Alexa-488), calcein（Calcein), cyanine -2 (Cyanine-2), fluorescein phosphoramidate（FAM), fluorescein isothiocynate（Fluorescein isothiocyanate, FITC), fluorite fluorescein X (FluorX), green fluorescent protein（GFP), red displacement green fluorescent protein（RsGFP), Oregon green -500 (Oregon Green-500) rhodamine -110 (Rhodamine 110), rhodamine are green（Rhodamine green) or SYBR green etc..Wherein, the optical filtering part 20 is 492nm for the wavelength that the excitation wavelength of correspondence 6- fluorescein phosphoramidates adjusts the filtered beam 21, and then the ripple of fluorescence 31 after being excited is longer than at 517nm, and carry out exciting 6- fluorescein phosphoramidates using 492nm wavelength, the optimal luminous efficiency of fluorescence 31 can be obtained；And if during using 5- fluorescein phosphoramidates as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 494nm, and obtaining the 518nm wavelength of fluorescence 31；If during using Oregon green -488 as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 496nm, and obtains the 524nm wavelength of fluorescence 31；If during using A Laisha -488 as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 495nm, and obtains the 520nm wavelength of fluorescence 31；If during using calcein as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 494nm, and obtains the 517nm wavelength of fluorescence 31；If during using cyanine -2 as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 489nm, and obtains the 506nm wavelength of fluorescence 31；If during using fluorescein phosphoramidate as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 488nm, and obtains the 508nm wavelength of fluorescence 31；If during using fluorescein isothiocynate as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 494nm, and obtains the 518nm wavelength of fluorescence 31；If during using fluorite fluorescein X as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 494nm, and obtains the 519nm wavelength of fluorescence 31；If during using green fluorescent protein as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 488nm, and obtains the 558nm wavelength of fluorescence 31；If during using red displacement green fluorescent protein as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 488nm, and obtains the 507nm wavelength of fluorescence 31；If during using Oregon green -500 as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 503nm, and obtains the 522nm wavelength of fluorescence 31；
If during using rhodamine -110 as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 496nm, and obtains the 520nm wavelength of fluorescence 31；If using rhodamine it is green as fluorescein stain 30 when, the wavelength of the filtered beam 21 is set as 502nm, and obtains the 527nm wavelength of fluorescence 31；If during using SYBR green as fluorescein stain 30, the wavelength of the filtered beam 21 is set as 497nm, and obtains the 520nm wavelength of fluorescence 31.Described above, in particular range of wavelengths, and then obtains the preferred launching efficiency of fluorescence 31 with the filtered beam 21 of accurate adjustment optical filtering part 20.In addition, due to the filter effect of optical filtering part 20, it controls light value in the filtered beam 21 before and after the crest in 15nm wavelength band, zero is just leveled off to more than the light value outside 15nm scopes, thus the wavelength as the wavelength of fluorescence 31 produced by above-mentioned fluorescein stain 30 all with filtered beam 21 is at a distance of more than 15nm, and measurement error problem when being not to cause to detect due to the wave-length coverage and the overlapping wavelengths of fluorescence 31 of filtered beam 21.In summary, due to the present invention exciting and detecting merely with the progress fluorescence 31 of blue light diode 10, and the wavelength and spectral range of filtered beam 21 are controlled through the adjustment of optical filtering part 20, and then obtain and efficient excite fluorescence 31, and by the wave-length coverage overlapping wavelengths not with fluorescence 31 for the filtered beam 21 for controlling optical filtering part 20, exclude error problem when detection module 40 measures the intensity of fluorescence 31.The present invention is elaborated above, described above, only the preferred embodiments of the present invention can not be used to limit the scope that the present invention is implemented.All equivalent variations made according to the present patent application scope and modification etc., in the patent covering scope that all should still belong to the present invention.