The present invention relates to the detection of fluorescence of biological molecules, and specifically, to the homogenization of and switching between the excitation light beams originating from different light emitting diodes (LEDs).
Many biological molecules of interest naturally fluoresce when excited by shorter-wavelength ultra-violet (UV) light. Traditionally, a fluorescence spectrophotometer measures the fluorescent light emitted from a sample at different wavelengths, after illumination with lamps; xenon arcs and mercury-vapor lamps. The light from an excitation source passes through a filter or monochromator and strikes the sample. A proportion of the incident light is absorbed by the sample, and some of the molecules in the sample fluoresce. A mercury vapor lamp is a line lamp, meaning it emits light near peak wavelengths. By contrast, a xenon arc has a continuous emission spectrum with nearly constant intensity in the range from 300-800 nm and a sufficient irradiance for measurements down to just above 200 nm. No monochromator is perfect at achieving complete blocking of the out of band light and it will transmit some stray light, that is, light with other wavelengths than the targeted. An advanced fluorometer uses UV light emitting diode (LED) light to excite the microorganisms in the sample into generating fluorescent energy (an auto-fluorescent signal). All these configurations with UV LED light only include one or two UV LEDs.
The Excitation Emission Matrix (EEM) is a specific measurement that is becoming more and more respected and widely used within the field of fluorescence spectroscopy, by exciting the sample with multiple different wavelengths and collecting and detecting fluorescent emission further, the fluorescence emission may be directed into a spectrophotometer to produce an excitation matrix. For the EEM, the light source may have a wavelength of, or have a wavelength in the range of 200 nm to 800 nm, such as, for example, 230 nm, 260 nm, 265 nm, 270 nm, 275 nm, 280 nm and 285 nm, but also at 300 nm˜420 nm in 5-10 nm wavelength separation.
In U.S. Pat. No. 8,651,695, a high intensity LED based lighting array has a ring assembly having a plurality of reflectors and LEDs. The ring assembly has a planar surface mounting each of the plurality of primary reflectors in perpendicular relation to a respective one of the plurality of light emitting diodes. A secondary diffuser is positioned on the ring to mix light from the LEDs to create a uniform light emission in a range of azimuth angles. Unfortunately, the coupling efficiency is very low due to reflector diffusion. In fact, the coupling efficiency may be less than 1 percent.
In U.S. Pat. No. 8,596,815, LED chips emit light within different wavelength ranges and are distributed laterally with respect to the axis over an area, where the LED chips have light emitting surfaces for emitting light in directions transverse to the area. An optical element adjacent to the light emitting surfaces of the LED chips in the at least one array collects and directs light emitted by the LED chips of the at least one array along the axis towards a target. Another embodiment is directed to a method for providing light having multiple wavelengths for fluorescent microscopy. The different wavelengths may have a different optical axis and the optical beam is not collimated because of the larger LED source.
United States patent publication number 20040189787 provides a rotating mirror to sequentially receive light from a plurality of solid state light emitters and to provide a time-multiplexed light output, but not for combining different wavelength with overlap spectrum. In addition, there is no rotation of the mirror.
In U.S. Pat. No. 6,760,506, an optical switch is provided for switching a light beam from at least one optical input to one of a plurality of outputs. In the optical switch all input beams have the same plane of incidence, and the mirror is only used for tuning light directions.
Finally, in U.S. Pat. No. 9,250,431, a high-power microscopy illumination system is disclosed, which includes a solid-state illumination source. A diffusing collection lens having a diffusing surface is configured to collect and diffuse light emission from the solid-state illumination source. An emitting surface is disposed substantially opposite the diffusing surface. An optical coupling element couples the light emission from the diffusing collection lens emitting surface along an optical axis to an optical output. The diffusing collection lens provides improved uniformity of illumination with direct coupling, without significant power loss. It is noted, however, that this system is for improving the output beam uniformity, not for improving the UV beam angular distribution consistency of different UV LED sources.
Advanced fluorometer systems for EEM analysis can improve the signal-to-noise ratio of the auto-fluorescence signal by using narrow bandwidth UV light emitting diode (LED) light instead of broadband lamp sources. Unfortunately, the combining of many LEDs within 5-10 nm wavelength separation using a traditional dichroic plate combiner, is not practical, and is very costly due to the requirements of sharp deep dichroic plate coating. This process is also very complicated, if not impossible. To date optical output of LEDs in UVB/UVC range are several times lower than UVA or color LEDs. In addition, optical coupling efficiency for combining multiple wavelength LED sources to application target would be low when using a traditional dichroic plate layout, because of reflective/absorptive losses encountered along the series of dichroic plates in the optical path. The dichroic filter losses at UVB/UVC wavelengths are considerably higher than the losses in the visible wavelength range due to substrate and coating material limitations. Therefore, there is a need to overcome these shortcomings of the prior art to improve the spectral quality of excitation signal while maintaining good optical coupling efficiency of the combined beam.
Embodiments of the present invention provide a multi-wavelength light source for switching a light beam with different wavelengths, wherein the light source covers a range of wavelengths. The light source contains a series of light emitting diodes (LEDs) arranged in a circular or semicircular pattern, where each LED is associated with one channel and has a different wavelength falling within the range of wavelengths. A rotational center mirror is provided and an engine that controls rotation of the rotational center mirror and switching speed of the mirror. Each LED has a different plane of incidence and a same incident ray angle, where a reflected output light beam of the light source is reflected in the same direction by the rotational center mirror, and wherein the rotational center mirror, as controlled by the engine, acts as a switch for light beams with different wavelengths that are received from the different LEDs. The series of LEDs arranged in a circular or semicircular pattern can be arranged on multiple levels where channels in layer one and layer two are offset to allow selection of one channel at a time, corresponding to its unique angular identification.
A dichroic beam combiner located in a beam path of a second channel associated with layer two can be provided serves as a dual function optical element, where the dichroic beam combiner has a cut-on/-off wavelength between the two sets of wavelength bands of layer one and layer two, so that the dichroic beam combiner allows light from the layer one channels to transmit through whereas the light from the layer two channels will be reflected so that both beams are combined in a final output. A single edge or multi-band dichroic beam splitter which specifically reflects wavelengths in the LEDs of the given layer but transmits other wavelengths can replace central rotating mirror to further aid in suppression of stray light in the output signal.
Other systems, methods and features of the present invention will be or become apparent to one having ordinary skill in the art upon examining the following drawings and detailed description. It is intended that all such additional systems, methods, and features be included in this description, be within the scope of the present invention and protected by the accompanying claims.
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification. The drawings illustrate embodiments of the invention and, together with the description, serve to explain the principals of the invention.
When multiple LEDs are used in an attempt to provide illumination that covers a range of spectrum of light, the LEDs traditionally provide light that is far apart in the spectrum, resulting in there being a number of discrete peaks within the spectrum covered by the multiple LEDs. However, in certain applications it is desirable to have a flat spectrum, such as a lamp spectrum, where there are little to no discrete peaks. An example of such an implementation is in the field of medical imaging where more and more markers are being used in the illumination of tissue and cells in general, resulting in the need for the emission of a greater spectrum of light without discrete peaks. Therefore, there is a need to have LEDs that can fill the gaps with other wavelengths so as to have a truer broadband illumination system where there are fewer peaks and valleys. The present system provides a continuum of wavelengths, as opposed to only specific wavelengths. The present system provides for multiple wavelengths of light to be provided and spaced very close together spectrally through the use of multiple LEDs in one or more array. For exemplary purposes, the present schematic description provides an example of using twelve LEDs with associated wavelengths that are spectrally adjacent is provided. It should be noted, however, that the present invention is not limited to twelve LEDs, but may instead be more or fewer LEDs.
Since the present system 2 contains multiple LEDs, where each LED has a specific wavelength and those wavelengths cover a range with all wavelengths within that range covered, the present system 2 provides a truer broadband system. This is even the case when the wavelengths are spaced very close together. This is ideal when there is a need to have a flat spectrum for the system, where multiple wavelengths need to be provided for by the system. An example of such use may be in the medical industry, where multiple fluorescence markers are provided and there is a need for a single system that can use multiple excitation wavelengths for detection.
In accordance with an alternative embodiment of the invention, the LEDs may instead be arranged in a semicircular arc so that there is not a full circular pattern of LEDs. In addition, while the present description provides the example of single LEDs arranged in a circular pattern, one having ordinary skill in the art would appreciate that there may instead be multiple LEDs that are in a single array 10A and each of the arrays of LEDs 10A-10H may be arranged in a circular or semi-circular arc pattern. In such a single array configuration, each of the LEDs within a single array may emit light within the same bandwidth. Alternatively, each of the LEDs within a single array (such as 10A) emits light having a different bandwidth. In such a further alternative embodiment, the multiple LEDs act as a single source with relatively wider bandwidth.
At the center of the circular LED arrangement is a rotational center mirror 50, which is controlled by a motor 60 (
It is preferred that an LED is turned on only when used by the system 2. Specifically, if light having a bandwidth of a first LED 10A is necessary, the system 2 provides power to the first LED 10A and directs the mirror 50 toward the first LED 10A. If light having a bandwidth of a fourth LED 10D is then required, the first LED 10A is turned off and the fourth LED 10D is turned on, with the mirror 50 then being turned to face the fourth LED 10D so that the associated light can be directed from the system 2. It should be noted that the timing of turning on an LED 10 and directing the mirror 50 may be such that the mirror 50 is directed first to face the LED 10 to be used, after which the LED to be used is turned on, or the LED may be turned on and then the mirror 50 turned. Either sequence will work in accordance with the present system 2 and associated method of use.
Referring to both
After collimation, the beam of light is received by the bandpass filter 40. The filter 40 may be a single bandpass filter or multiple bandpass filters stacked to achieve intended levels of rejection of the out-of-spectral-band light from the excitation source. Although each of the LEDs 10A-10H have their unique incident planes, a selected channel has the same incident angle with respect to the rotating mirror 50 when the mirror 50 is aligned to that channel, resulting in the reflected output light beam from the mirror 50 having the same direction and target.
The system 100 of
Channel seven, as illustrated by
A dichroic beam combiner, or filter 70, is located in a beam path of channel 1. The dichroic filter 70 has a cut-on/-off wavelength between the two sets of wavelength bands of top and bottom layers. The dichroic allows light from the top layer channels to transmit through whereas the light from bottom channels will be reflected so that both beams are combined in the final output.
As illustrated by
The plate and sensor assembly is attached to the motor axis and rotates with the dichroic mirrors. The reverse orientation with respect to the dichroic allows it to compensate for the reflected beam walk off from bottom channels.
It should be noted that the rotational center mirror 50 can be used in switching Laser with different wavelengths. In such an embodiment, the LEDs may be Laser LEDs or the LEDs may be replaced by lasers having the characteristics of the LEDs as mentioned herein.
It is noted that the present system and method is not limited to one or two levels of circular arrangement of LEDs.
While this specification contains many specific implementation details, these should not be construed as limitations on the scope of any inventions or of what may be claimed, but rather as descriptions of features specific to particular embodiments of particular inventions. Certain features that are described in this specification in the context of separate embodiments can also be implemented in combination in a single embodiment. Conversely, various features that are described in the context of a single embodiment can also be implemented in multiple embodiments separately or in any suitable sub-combination. Moreover, although features may be described above as acting in certain combinations and even initially claimed as such, one or more features from a claimed combination can in some cases be excised from the combination, and the claimed combination may be directed to a sub-combination or variation of a sub-combination. Other implementations are within the scope of the claims.
Number | Date | Country | |
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Parent | PCT/US2022/075625 | Aug 2022 | WO |
Child | 19062641 | US |