WO2020086156A1

WO2020086156A1 - Microscope with led illumination assembly

Info

Publication number: WO2020086156A1
Application number: PCT/US2019/049392
Authority: WO
Inventors: Congliang Chen
Original assignee: Congliang Chen
Priority date: 2018-08-31
Filing date: 2019-09-03
Publication date: 2020-04-30
Also published as: WO2020086156A9; CN112955790A

Abstract

A microscope system, the system comprising of a multi-channel LED lighting array comprising one or more optical redirection nodes, the optical redirection nodes comprised of a front face, a rear face, a top face, a bottom face, and two side faces; one or more light emitting diodes, each positioned to face one of the front face, rear face, top face, or bottom face of one of the one or more optical redirection nodes; and one or more collimating lenses, each positioned between one of the one or more light emitting diodes and one of the one or more optical redirection nodes; and a microscope assembly comprising an objective, one or more magnifying lenses, an optical path, and a specimen stage.

Description

MICROSCOPE WITH LED ILLUMINATION ASSEMBLY

TECHNICAL FIELD

[0001 ] This invention relates generally to a microscopy and spectrophotometry system, and more specifically fluorescent imaging systems.

BACKGROUND ART

[0002] Fluorescence microscopy allows the study of material that can be made to fluoresce, either in its natural form (termed primary or auto fluorescence) or when treated with chemicals capable of fluorescing (known as secondary fluorescence). The fluorescence microscope, and fluorescence microscopy is typically considered an important tool in cellular biology.

[0003] The phenomenon of fluorescence occurs where a sample is illuminated with a wavelength of light the fluorescing light produced by the sample typically has longer wavelengths than the excitation light, a phenomenon known as the Stokes shift (or photoluminescence). Shorter wavelength light in the UV part of the spectrum tends to have more photonic energy (when considering the particle nature of light) than longer wavelength infrared light. For example, when a sample is illuminated by ultraviolet radiation (purple) it may be absorbed by an electron in a particular atom, exciting and elevating the electron to a higher energy level. Subsequently, the excited electron relaxes to a lower level and the transition causes the release of energy in the form of a lower-energy (red) light in the visible light region.

[0004] If the emission of light continues for typically up to a few seconds after the exposure to excitation energy (light) is discontinued, the phenomenon is known as

phosphorescence. Fluorescence is the light emission that occurs during the absorption of the excitation light. The time interval between the absorption of excitation light and emission of re radiated light in fluorescence is typically finite but extraordinarily short in duration, usually less than a millionth of a second.

[0005] Fluorescence microscopy accordingly includes illumination equipment allows the selections of excitation wavelengths, and equipment to image and record the emission

wavelengths fluoresced by the material. Accordingly devices that improve and efficiently provide illumination as well as the capture of images in fluorescence microscopy would be useful. DISCLOSURE OF INVENTION

[0006] The following presents a simplified summary of the disclosure in order to provide a basic understanding to the reader. This summary is not an extensive overview of the disclosure and it does not identify key/critical elements of the invention or delineate the scope of the invention. Its sole purpose is to present some concepts disclosed herein in a simplified form as a prelude to the more detailed description that is presented later.

[0007] This invention relates generally to a microscopy and spectrophotometry system, and more specifically to a fluorescent imaging system. Specific details of certain examples of the invention are set forth in the following description and in the figures to provide a thorough understanding of such examples. The present invention may have additional examples, may be practiced without one or more of the details described for any particular described example, or may have any detail described for one particular example practiced with any other detail described for another example.

[0008] A microscope with a LED illumination assembly may be implemented in an exemplary Stereo Fluorescent Microscope with a LED cartridge module light source. The present invention includes a microscope with a LED illumination assembly that significantly reduces light intensity loss, allows for easy changing of LEDs (and accordingly the wavelengths of light applied), and allows for improved uniformity of light intensity.

[0009] In an example of the device, a group of three LED assemblies, each projecting a different wavelength of light (though they may also be the same wavelength if desired) is arranged around a cubic optical prism. The optical prism is designed to bend an approaching beam from the optical prism’s left or right faces ninety (90) degrees to exit out of the front face. A beam incoming from the optical prism rear face exits the front face without deflection. One LED assembly is positioned facing the left face, one the right face, and one the rear face. Each LED Assembly has a collimating lens disposed between its LED and the cubic optical prism, which converts the incoming light beam into a column. The collimated beam from the left face facing LED enters into the cubic optical prism and is diverted to exit out the front face, the collimated beam from the right face facing LED enters into the cubic optical prism and is diverted to exit out the front face, and the collimated beam from the rear face facing LED passes through the cubic optical prism to exit out the front face. Light from all three LEDs thus passes through near identical optical paths to the objective. Accordingly the LEDs may be similarly sized, as they all typically encounter similar system losses. In this example, the cubic optical prism reduces the light passing through it by 50%, but as a result 50% of each LED's light passes through the crystal.

[001 0] As a result, the amount of light intensity lost from each LED as its light passes through the cubic optical prism is equal and the amount of light that reaches the objective from each LED is therefore roughly equal. The benefit of such an arrangement is that the amount of power needed to utilize each LED can be roughly equal instead of current arrangements where the LEDs and cubic optical prisms are arranged in a chain and thus the LEDs towards the back of the chain must be many times more intense in order to provide the same amount of light to the objective.

[001 1 ] In some examples, the cubic optical prisms are optical redirection nodes that may or may not be substantially flat pieces of optical glass configured to redirect or split beams of light of varying wavelengths, or beam splitters.

[001 2] The present invention also allows for the LEDs to be easily installed, removed, and/or replaced without the need for recalibration or calculation.

[001 3] The LEDs in the present invention may draw power from a variety of sources, including, but not limited to, internal batteries, direct connections, or through a connection to a power-connected baseplate or other securing mechanism.

[0014] Many of the attendant features will be more readily appreciated as the same becomes better understood by reference to the following detailed description considered in connection with the accompanying drawings.

BRIEF DESCRIPTION OF DRAWINGS

[001 5] The present description will be better understood from the following detailed description read in light of the accompanying drawings, wherein :

[001 6] FIG. 1 illustrates fluorescence microscopy in the study of material that can be made to fluoresce.

[001 7] FIG. 2 shows a block diagram of a conventional LED lighting assembly that may be used in conventional fluorescence microscopy; [001 8] FIG. 3 shows a perspective view of a first example of the new multichannel LED lighting system having three light sources;

[001 9] FIG. 4 shows a block diagram of the multichannel LED lighting system having three light sources;

[0020] FIG. 5 shows a perspective view of an example of a multichannel LED lighting system having five light sources;

[0021 ] FIG. 6 shows a perspective view of an example of a multichannel LED lighting system having seven light sources;

[0022] FIG. 7 shows an exploded, perspective view of an example of a microscope with multichannel LED lighting system having seven light sources;

[0023] FIG. 8 cross section D-D of FIG.6

[0024] FIG. 9 is a detail view of Detail E of FIG. 8

[0025] FIG. 1 0 shows a front perspective view of an example of a multichannel LED lighting system LED assembly;

[0026] FIG. 1 1 shows a rear perspective view of an example of a multichannel LED lighting system LED assembly;

[0027] FIG. 1 2 shows an exploded view of an example of a multichannel LED lighting system LED assembly;

[0028] FIG. 1 3 shows a first perspective view of an alternative example of a multichannel

LED lighting system;

[0029] FIG. 1 4 shows a second perspective view of an alternative example of a multichannel LED lighting system;

[0030] FIG. 1 5 shows a third perspective view of an example of a multichannel LED lighting system;

[0031 ] FIG. 1 6 shows a perspective view of an example of a microscope including a multichannel LED lighting system;

[0032] FIG. 1 7 shows a perspective view of a portion of the example shown in FIG. 1 6 showing the microscope stage, optical path, including a multichannel LED lighting system;

[0033] FIG. 1 8 shows a side view of a portion of the example shown in FIG. 1 6 showing the microscope stage, optical path, and multichannel LED lighting system of FIG. 1 1 ; [0034] FIG. 1 9 describes a process for providing multiple fluorescence images of a sample in a single image.

[0035] FIG. 20 illustrates an exemplary computing environment in which the imaging system including LED light source control described in this application, may be implemented.

[0036] Like reference numerals are used to designate like parts in the accompanying drawings.

BEST MODE FOR CARRYING OUT THE INVENTION(S)

[0037] This invention relates generally to a microscopy and spectrophotometry system, and more specifically to a fluorescent imaging system. Specific details of certain examples of the invention are set forth in the following description and in FICs. 1 -20 to provide a thorough understanding of such examples. The present invention may have additional examples, may be practiced without one or more of the details described for any particular described example, or may have any detail described for one particular example practiced with any other detail described for another example.

[0038] As used herein and unless otherwise indicated, the terms“a” and“an” are taken to mean“one”,“at least one” or“one or more”. Unless otherwise required by context, singular terms used herein shall include pluralities and plural terms shall include the singular.

[0039] Unless the context clearly requires otherwise, throughout the description and the claims, the words‘comprise’,‘comprising’, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of“including, but not limited to”. Words using the singular or plural number also include the plural and singular number, respectively. Additionally, the words“herein,”“above,” and“below” and words of similar import, when used in this application, shall refer to this application as a whole and not to any particular portions of the application.

[0040] The detailed description provided below in connection with the appended drawings is intended as a description of the present examples and is not intended to represent the only forms in which the present example may be constructed or utilized. The description sets forth the functions of the example and the sequence of steps for constructing and operating the example. However, the same or equivalent functions and sequences may be accomplished by different examples.

[0041 ] FIG. 1 illustrates fluorescence microscopy in the study of material that can be made to fluoresce. A sample 6 is illuminated with excitation light 1 1 , causing such a receptive sample to fluoresce 1 2 at a differing wave length which may be observed through a microscope.

[0042] Typically a light source 1 emits monochromatic light 2 that is filtered with an excitation filter 3 to pass light of a given wavelength 1 4 desirable to test the sample 6 placed on a microscope stage 7. The selected wavelength light 1 4 is directed to a dichroic mirror 4 where it passes through an objective 5 to illuminate a sample 6. If the sample has the proper

characteristics it may fluoresce at a different wavelength 1 2, which is directed through the objective 5, passes through a dichroic mirror 4, then an emissions filter 8, an ocular 9, and then a finally to a detector 1 0.

[0043] The present examples described below provide an improved way of providing light to illuminate a sample with selective wavelength light, as past set ups have been prone to loss of light in the signal path and high power consumption.

[0044] FIG. 2 shows a block diagram of a conventional LED lighting assembly that may be used in conventional fluorescence microscopy. Current systems 200 such as the four light source system shown typically operate in cascade. Such systems may include one or more cascaded cubic optical prism 206, 21 2 , 21 8 each with a front face 234, left face 236, right face 238, and rear face 249, and a plurality of light sources 226, 228, 230, 232 , here shown as LEDs. Each light source 226, 228, 230, 232 has a respective collimating lens 204, 208, 21 4, 220 between the LEDs 202, 208, 21 4, 220 and the cubic optical prism, which converts the incoming light beam from the LED into a collimated output input to the optical prism it is coupled to.

[0045] Initially, in providing a first and second wavelength, two light sources 226, 228 of the same or different wavelengths may be arranged with one 226 pointing at the rear face, and one 228 at the right face.

[0046] In providing for the first color (wavelength) the collimated beam from the rear face facing light source 226 passes through the cubic optical prism 206 to exit out the front face

[0047] In providing the second wavelength. The right facing light source 228 enters into the cubic optical prism 206 and is diverted by action of the prism to exit out the front face. [0048] In order to add a third color of light a second cubic optical prism 21 2 is added to the system wherein the light from the first cubic optical prism 204 leaves its front face and enters the rear face of the second cubic optical prism 21 2 and then leaves through the front face of the second cubic optical prism 21 2. A third light source 230 is positioned such that it faces the right face of the second cubic optical prism 21 2 , and light from that light source 1 02 enters the second cubic optical prism 21 2 and is diverted to exit out the front face.

[0049] In order to provide a fourth wavelength a third prism 21 8 is added in cascade. The light leaving the second cubic optical prism 21 2 then enters the rear face of the third cubic optical prism 21 8 and passes out through its front face. A fourth light source 232 is positioned such that it faces the right face of the third cubic optical prism 21 8, and light from that light source 232 enters the third cubic optical prism 21 8 and is diverted to exit out the front face.

[0050] The combined beams of light pass through the optical path formed by the prisms

206, 21 2 , 21 8 to reach an objective 224. At this the objective, the light from the fourth light source 232 is at 50% intensity, the light from the third light source 230 is at 25% intensity, and the light from the first 226 and second 228 light sources are each at 1 2.5% intensity. As such top provide uniform illumination, the first two light sources 226, 228 would typically be at least four (4) times as intense as the final light source 232 in order to provide the same amount of intensity on the objective as the fourth light source 1 02. Any additional cubic optical prism and LED pairs would thus require proportional increases in intensity. It would be desirable to have an optical configuration to utilize light sources of similar output and power consumption to aid in providing for flexibility in setting up desired light sources in any given test set up. The present invention thus improves on the state of the art by ensuring that any intensity loss is either uniform or significantly easier to compensate for.

[0051 ] In some alternative versions of the current art, several light sources are

simultaneously illuminated with a mechanism provided (not shown) to only pass one source at a time. In such a configuration the mechanism rapidly changes which light source is being transmitted to give the illusion of several light sources having the same intensity. However, this mechanical setup comes with the drawback of not combining well with the refresh rates of digital displays, which can result in poor viewing. The present invention solves this problem by eliminating the rapidly changing blocking mechanism in favor of having the light wavelengths actually broadcast at the target at the same time.

[0052] The microscope with an LED illumination assembly described below is typically implemented as Automated Fluorescence Microscope that includes the LED illumination assembly. The automated microscope described below is unique in that it is an integrated and modular assembly that may be easily moved and set up again with a minimum of adjustment. Current automated systems tend to be assembled from various components that are not easily moved once set up and adjusted in a given location. Also when initially installed a lengthy calibration and adjustment procedure typically must be performed before the imaging set up may be used. The microscope with LED illumination assembly tends to provide a truly integrated microscopy solution that is portable and easy to use.

[0053] In addition the LED illumination assembly is especially useful because it allows for a variety of wavelengths of light to be projected on an objective at equal (or nearly equal) intensities, thus allowing users of fluorescing dyes to mark numerous targets at the same time using different wavelength absorbing dyes without becoming unable to see certain dyes due to lost LED intensity.

[0054] FIG. 3 shows a perspective view of a first example of the new multichannel LED lighting system having three light sources 300. The LED illumination assembly 300 includes a base plate 304 and one or more LED light assemblies 306, 31 0, 31 2. Each of which include a light source 302 (that are similarly constructed, but may be of differing wavelengths) coupled to a collimating lenses 307. The LED light assemblies 306, 31 0, 31 2 are disposed around a cubic optical prism 308. As well as providing mechanical support for the LED/lens pieces 302 , 307 the base plate 304 may also provide electrical connections for powering and controlling the illumination produced by the assembly.

[0055] Light is produced by each of the LED light assemblies 306, 31 0, 31 2 typically with each module producing a different wavelength. The varied light sources may then be combined using the cubic optical prism 308 before exiting the device to illuminate a sample. The combined light beam is typically put to use in the field of fluorescence microscopy or any other suitable applicable endeavor. The system 300 is designed so that one or more light sources 302 can be readily installed and /or replaced by the end user without additional calibration of the system 300. In the example shown the combination of three exemplary light sources is shown. However, those skilled in the art will realize that the principles described herein may be used to combine any number of light sources, of any variety of wavelengths.

[0056] The assembly shown includes three LED light assemblies 306, 31 0, 31 2 which are positioned to point towards different faces of the cubic optical prism 308. The light leaves through the face of the cubic optical prism 308 that does not have an LED light assembly 306,

31 0, 31 2 pointed towards it. In some alternative examples, additional LED light assemblies (not shown) may be positioned so that they also face the top and/or bottom faces of the cubic optical prism 308 which may be configured to redirect their light out the same side as lateral facing sources 302.

[0057] The one or more light sources 302 may be LEDs or equivalently other forms of light source including, but not limited to, incandescent lights, mercury lamps, lasers or the like. The collimating lenses 307 are conventional in function. The cubic optical prism 308 also functions conventionally as is known to those skilled in the art, and may include a variety of different components configured to allow light to pass through and /or be deflected. The cubic optical prism 308 may be comprised of multiple components or smaller prisms to achieve the same result.

[0058] FIG. 4 shows a block diagram of the multichannel LED lighting system having three light sources 300. This diagram shows component assembly and optical path of a microscope with multichannel LED lighting system. In an example of the device, a group of three light sources 306, 31 0, 31 2, each projecting a different wavelength of light, though they may also be the same wavelength if desired, are arranged around a cubic optical prism 308 designed to bend an approaching beam from its left 436 or right 438 faces ninety (90) degrees to exit out of the front face 434, while allowing any beam incoming from its rear face 449 to exit the front face 434 without deflection.

[0059] Light source 306 is positioned facing the left face, Light Source 31 0 faces the right face, and light source 31 2 faces the rear face. Each light has a collimating lens 307 between it and the cubic optical prism 308, which converts the incoming light beam into a column. The collimated beam from the left face facing light source 306, 31 0 enters into the cubic optical prism 308 and is diverted to exit out the front face 434, the collimated beam from the right face facing light source 31 0 enters into the cubic optical prism 308 and is diverted to exit out the front face 434, and the collimated beam from the rear face facing light source 31 2 passes through the cubic optical prism 308 to exit out the front face. Light from all three light sources 306, 31 0, 31 2 passes through equal loss optical paths 21 0 to reach the objective 41 2 with each source typically having equal light intensity generated at their respective wavelengths. Additional light sources may be added by cascading additional optical prisms. Cascading may be achieved by attaching one or more additional prisms together with conventional optically transparent adhesive to form a prism with additional light inputs. Then additional prisms may be disposed on each side (left and right faces) of the extended prism.

[0060] FIG. 5 shows a perspective view of an example of a multichannel LED lighting system having five light sources 500. The system 500 includes one or more light sources 302 coupled to a base plate 504 and contained in one or more LED light assemblies 506, 508, 51 0,

51 2 , 51 4, which are comprised of the light source 1 02 and one or more collimating lenses 307; with each of the LED light assemblies arranged around a cubic optical prism 520. In some examples, electrical power and communication may be provided through the base plate 504, from a system controller, simultaneously providing power to each of three LED light assemblies 506, 508, 51 0, 51 2, 51 4. Light is produced by each of the LED light assemblies 506, 508, 51 0, 51 2 ,

51 4, each of a different wavelength.

[0061 ] The varied light sources 506, 508, 51 0, 51 2 , 51 4 are combined using a cubic optical prism 520 before exiting the device through an exit port. In this example the cubic optical prism is comprised of two of the previously described cubic optical prisms (308 of FIG. 3) attached by an optically transparent adhesive.

[0062] The combined light beam may be put to useful purpose in the field of fluorescence microscopy or any other suitable applicable endeavor. The system 500 is designed so that one or more light sources 506, 508, 51 0, 51 2 , 51 4 can be readily installed and /or replaced by the end user without additional calibration of the system 500.

[0063] FIG. 6 shows a perspective view of an example of a multichannel LED lighting system having seven light sources 600. The system 600 includes one or more light sources 620, 622 , 624, 626, 628, 630, 632 coupled to a base plate 604 each having a LED light source 302 and a collimating lenses 307. Light typically produced by each of the LED light assemblies may be of a different wavelength. Each of the LED light assemblies may be arranged around a cubic optical prism 602.

[0064] The varied light sources are combined using a cubic optical prism 602 before exiting the device through an exit port. In this example the cubic optical prism 602 is comprised of three cubic optical prisms constructed as previously described (308 of FIG. 3) and attached to each other by an optically transparent adhesive.

[0065] In some examples, electrical power and communication may be provided through the base plate 604, from a system controller (not shown), simultaneously providing power and control of each of seven LED light assemblies 620, 622 , 624, 626, 628, 630, 632.

[0066] The combined light beam is then put to useful purpose in the field of fluorescence microscopy or any other suitable applicable endeavor. The system 600 is designed so that one or more light sources 620, 622 , 624, 626, 628, 630, 632 can be readily installed and /or replaced by the end user without additional calibration of the system 600.

[0067] FIG. 7 shows an exploded, perspective view of an example the multichannel LED lighting system having seven light sources (600 of FIG. 60). There are seven light sources (302 of FIG. 3) incorporated into the seven LED light assemblies 620, 622 , 624, 626, 628, 630, 632. The LED light assemblies may be optically coupled to three cubic optical prisms 308 to form a unit 602.

[0068] A housing 704 provides mounting and alignment of the modules 620, 622, 624,

626, 628, 630, 632 , and the prisms 602. The housing 704 may be mechanically coupled to the base plate 604 with screws or equivalent. The base plate 604 provides electrical power to the plurality of LED light assemblies 620, 622, 624, 626, 628, 630, 632 , which are mechanically, and removably coupled to the housing 704.

[0069] The triple cubic optical prism 602 is composed of three identical cubic optical prisms 308 which are bonded together with optically clear adhesive. A single cubic optical prism 1 08 is constructed such that three light beams, each of different wavelengths, can enter the cubic optical prism 1 08 from each of three sides, combine, and then exit the fourth side. The combined light beam then exits the device through exit port 702.

[0070] The LED light assemblies 620, 622 , 624, 626, 628, 630, 632 , provide the source of light of a particular wavelength, then focuses and collimates the light before the light enters one side of the triple cubic optical prism disposed laterally to a given LED light assembly. In some examples, the light intensity is typically only lost when the light passes out of a cubic optical prism 308 and into air. Accordingly, the only light intensity loss occurs when light enters the triple cubic optical prism 602 , meaning the light from any given light source is advantageously only reduced in intensity once.

[0071 ] FIG. 8 shows the cross section D-D of FIG.6. The housing 704 provides support for the LED light assemblies 620, 622, 624, 626, 628, 630, 632 that are inserted into it. The housing provides a reliable optical coupling of light to the prisms (not shown) disposed within the housing. In addition alignment in the housing is aided by a chamfer 802 disposed on the edge of each LED light assembly.

[0072] FIG. 9 is a detail view of Detail E of FIG. 8. The triple cubic optical prism (602 of

FIG. 6) is secured in the housing 704 by location features 62 which contact the prism at a plurality of locations typically around the base of the prism. A plurality of centering features 61 disposed on the housing, and which are conically-shaped, are utilized to accurately center and aim the plurality of LED light assemblies (helping to ensure perpendicularity of the respective light beams with the prism faces). The centering features 61 contact adjacent features on the LED light assemblies once the light assemblies are fully inserted into the housing 704.

[0073] FIG. 1 0 shows a front perspective view of an example of a multichannel LED lighting system LED assembly. FIG. 1 1 shows a rear perspective view of an example of a multichannel LED lighting system LED assembly. The following description relates to these two figures.

[0074] In some examples, the system may be made up of one or more LED light assemblies

306, 31 0, 31 2, 406, 508, 51 0, 51 2, 51 4, 620, 622, 624, 626, 628, 630, 632 , each configured to or capable of being configured to project a different wavelength of light, typically by providing a LED light source 302 of a desired wavelength. The LED light assemblies may be made up of the combination of an LED light source 302 (or similar light source) and a lens set 307. In some examples they may be further comprised of an integrated collimating lens to collimate the light as it leaves the source.

[0075] The LED light assemblies are further comprised of the light sources 302 and the collimating lens 307 assemblies that are combined into the single LED light assembly for ease of replacement and to ensure that the LED is properly aligned. An integral connector 1 1 02 provides power, LED control, and module identification connections.

[0076] FIG. 1 2 shows an exploded view of an example of a multichannel LED lighting system LED assembly. In some examples, the LED light assembly 306, 31 0, 31 2, 406, 508, 51 0, 51 2 , 51 4, 620, 622, 624, 626, 628, 630, 632 includes of a housing 1 202 which contains and/or secures all of the various components. The LED focus adjustment assembly 1 204 contains the LED light source. The LED focus adjustment assembly 1 204 is inserted into the barrel of the LED assembly housing 1 202. The LED focus and anti-rotation set screw 1 206 is inserted through the side of the housing 1 202 and secured into the side of the LED focus adjustment assembly 1 204. This set screw serves to first secure the focal length adjustment of the LED and second to keep the LED focus adjustment assembly 1 204 from rotating in the barrel of the LED assembly housing 1 202. The concentricity of the LED with the lens set assembly 1 208 is adjusted by moving, perpendicular to the centerline axis, the LED focus adjustment assembly within the barrel of the LED assembly housing 1 202. Once the desired alignment is achieved, the three LED centering set screws 1 21 0 are secured against the angled conical face of the LED focus adjustment assembly 902. The lens set assembly 1 208 is installed in the LED assembly housing 1 202 from the opposite end of part. The lens set assembly may be secured with a retaining ring 1 21 2. A light filter 1 21 4 is also installed and secured with the retaining ring. The electrical connection 1 21 6 to the LED light assembly 1 202 is secured with screws, and is wired to the LED mount.

[0077] FIG.S 1 3-1 5 show various perspective views of an alternative example of a multichannel LED lighting system 1 300. In this example the base has been modified to couple to a microscope assembly. The system 1 300 is comprised of one or more light sources 1 02 attached to a base plate 1 04 and contained in one or more LED light assemblies 1 306, arranged around a cubic optical prism 1 308. In some examples, electrical power and communication is provided through the base plate 1 31 0, from a system controller (not shown), simultaneously providing power to each of three LED light assemblies 1 306. Light is produced by each of the LED light assemblies 1 306, each typically of a different wavelength. The varied light sources are then combined using a cubic optical prism 1 308 before exiting the device through an exit port 1 31 2. The combined light beam is then put to useful purpose in the field of fluorescence microscopy or any other suitable applicable endeavor. The system 1 300 is designed so that one or more light sources 1 306 can be readily installed and/or replaced by the end user without additional calibration of the system.

[0078] As previously described the LED light assemblies are configured to point towards different faces of the cubic optical prism 1 308. The light leaves through the face of the cubic optical prism 1 31 2 that does not have an LED light assembly 1 306 pointed towards it. In some examples, LED light assemblies 1 306 may be positioned so that they also face the top and /or bottom faces of the cubic optical prism 1 308 which may be configured to redirect their light out the same side as lateral facing prism faces.

[0079] FICs. 1 6-1 8 show an example of a microscope system utilizing the LED light assemblies of FIGs 1 3-1 5. The following description is in relation to FIGs. 1 6-1 8. The LED lighting system 1 300 may be integrated into a microscope 1 600. The microscope imaging portions may be of a variety of types including, but not limited to, stereoscopic or inverted.

[0080] This innovation exploits a merged technique through which multiple and varied wavelengths of light sources, produced by a plurality of varied wavelength LED sources, are collimated and then combined to produce a single collimated light output which can be used for productive endeavors. This system 1 600 is designed so that the individual light sources can be readily installed and /or replaced by the end user without additional calibration of the system 1 600. When employed in a microscope system the LED light assemblies provide minimal proportion intensity loss between light sources, ensuring that no single wavelength outshines the others to a degree that a user might find it difficult to spot certain types of fluorescence.

[0081 ] The microscope imaging system utilized 1 600 may be of a variety of types, including, but not limited to, a compound light microscope, stereo microscope, and /or digital microscope, but is not limited solely to such examples and the invention may be employed in a light-based imaging system.

[0082] In the present example, the system includes an exemplary five lite LED light assembly 1 300, or equivalent including any number of light sources, coupled to a 90 degree reflector assembly 1 700 which is in turn coupled to a focus adjustment tube 1 71 2 , going through a mounting plate 1 702. Also coupled to the mount plate 1 702 is a light tube 1 704, with a ninety degree mirror and filter assembly 1 706 that directs light to a stage 1 71 5. The multi-channeled light beam then travels upward through the objective lens to a specimen in a specimen tray resting on the stage 1 71 5. The light image then travels downward, through the fluorescence interference filter block 1 706, to the eye piece/detector (not shown). In some alternative examples, the eye piece/detector is a traditional glass lens eyepiece 1 606, while in other examples the eyepiece may be a charge-coupled device or other digital display.

[0083] FIG. 1 9 describes a process for providing multiple fluorescence images of a sample in a single image. First the number of illumination sources to be used is determined 1 902. At 1 904 a first illumination source of a plurality of illumination sources is provided for a period of time to create a first image 1 906, and then switched off 1 908. At 1 908 the process is repeated going back to block 1 904 for the next illumination source of the plurality of illumination sources until all sources selected have been activated in sequence 1 91 0.

[0084] The images produced may be displayed in real time having the effect of producing a composite fluorescent image of the multiple fluorescence’s created by each light source illuminating the sample. 1 91 2. In alternative examples the illumination parameters any number of illumination sources outputting a desired wavelength, or range of wavelengths of light, illumination time of individual sources, and the like may be sequentially applied to the rotating or repeating images 1 91 4. Accordingly a composite image created by this method presents the fluorescence created by each illumination source as a single image as perceived by the viewer.

[0085] FIG. 20 illustrates an exemplary computing environment 2000 in which the imaging system including LED light source control described in this application, may be implemented. Exemplary computing environment 2000 is only one example of a computing system and is not intended to limit the examples described in this application to this particular computing environment.

[0086] For example the computing environment 2000 can be implemented with numerous other general purpose or special purpose computing system configurations. Examples of well- known computing systems, may include, but are not limited to, personal computers, hand-held or laptop devices, microprocessor-based systems, multiprocessor systems, set top boxes, gaming consoles, consumer electronics, cellular telephones, PDAs, and the like.

[0087] The computer 2000 includes a general-purpose computing system in the form of a computing device 2001 . The components of computing device 2001 can include one or more processors (including CPUs, GPUs, microprocessors and the like) 2007, a system memory 2009, and a system bus 2008 that couples the various system components. Processor 2007 processes various computer executable instructions, including those to control the operation of the microscope including the illumination provided by the LED light sources, and to display the images, as well as to control the operation of computing device 2001 and to communicate with other electronic and computing devices (not shown). The system bus 2008 represents any number of several types of bus structures, including a memory bus or memory controller, a peripheral bus, an accelerated graphics port, and a processor or local bus using any of a variety of bus architectures.

[0088] The system memory 2009 includes computer-readable media in the form of volatile memory, such as random access memory (RAM), and /or non-volatile memory, such as read only memory (ROM). A basic input/output system (BIOS) is stored in ROM. RAM typically contains data and/or program modules that are immediately accessible to and /or presently operated on by one or more of the processors 2007.

[0089] Mass storage devices 2004 may be coupled to the computing device 2001 or incorporated into the computing device by coupling to the buss. Such mass storage devices 2004 may include a magnetic disk drive which reads from and writes to a removable, nonvolatile magnetic disk (e.g., a“floppy disk”) 2005, or an optical disk drive that reads from and/or writes to a removable, non-volatile optical disk such as a CD ROM or the like 2006. Computer readable media 2005 , 2006 typically embody computer readable instructions, data structures, program modules and the like supplied on floppy disks, CDs, portable memory sticks and the like.

[0090] Any number of program modules can be stored on the hard disk 201 0, Mass storage device 2004, ROM and /or RAM 200 9, including by way of example, an operating system, one or more application programs, other program modules, and program data. Each of such operating system, application programs, other program modules and program data (or some combination thereof) may include an example of the systems and methods described herein.

[0091 ] A display device 2002 can be connected to the system bus 2008 via an interface, such as a video adapter 201 1 . A user can interface with computing device 702 via any number of different input devices 2003 such as a keyboard, pointing device, joystick, game pad, serial port, and/or the like. These and other input devices are connected to the processors 2007 via input/output interfaces 201 2 that are coupled to the system bus 2008, but may be connected by other interface and bus structures, such as a parallel port, game port, and /or a universal serial bus (USB).

[0092] Computing device 2000 can operate in a networked environment using connections to one or more remote computers through one or more local area networks (LANs), wide area networks (WANs) and the like. The computing device 2001 is connected to a network 201 4 via a network adapter 201 3 or alternatively by a modem, DSL, ISDN interface or the like.

[0093] One skilled in the art will recognize that the herein described components (e.g., operations), devices, objects, and the discussion accompanying them are used as examples for the sake of conceptual clarity and that various configuration modifications are contemplated.

Consequently, as used herein, the specific exemplars set forth and the accompanying discussion are intended to be representative of their more general classes. In general, use of any specific exemplar is intended to be representative of its class, and the non-inclusion of specific components (e.g. , operations), devices, and objects should not be taken limiting.

[0094] With respect to the use of substantially any plural and /or singular terms herein, those having skill in the art can translate from the plural to the singular and /or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations are not expressly set forth herein for sake of clarity.

[0095] In some instances, one or more components may be referred to herein as

“configured to,”“configured by,”“configurable to,”“operable/operative to,”“adapted/adaptable,” “able to,”“conformable/conformed to,” etc. Those skilled in the art will recognize that such terms (e.g.“configured to”) generally encompass active-state components and /or inactive-state components and /or standby-state components, unless context requires otherwise.

[0096] While particular aspects of the present subject matter described herein have been shown and described, it will be apparent to those skilled in the art that, based upon the teachings herein, changes and modifications may be made without departing from the subject matter described herein and its broader aspects. It will be understood by those within the art that, in general, terms used herein, are generally intended as“open” terms (e.g. , the term“including” should be interpreted as“including but not limited to,” the term“having” should be interpreted as “having at least,” the term“includes” should be interpreted as“includes but is not limited to,” etc.). [0097] Accordingly, the scope of the invention is not limited by the disclosure of these preferred and alternate examples. Instead, the invention should be determined by reference to the claims that follow.

Claims

1 . A multi-channel LED lighting system, the system comprising:

one or more optical redirection nodes, the optical redirection nodes comprised of a front face, a rear face, a top face, a bottom face, and two side faces;

one or more light sources, each positioned to face one of the front face, rear face, top face, or bottom face of one of the one or more optical redirection nodes; and

one or more collimating lenses, each positioned between one of the one or more light sources and one of the one or more optical redirection nodes.

2. The system of claim 1 , wherein the one or more light sources are each positioned substantially perpendicular to one of the front face, rear face, top face, bottom face, or side face of the one or more optical redirection nodes.

3. The system of claim 1 , wherein there are at least two optical redirection nodes arranged such that the rear face of one is parallel to the front face of the other.

4. The system of claim 1 , wherein the one or more light sources each are configured to emit a different wavelength of light.

5. The system of claim 4, wherein the wavelengths of light emitted by the one or more light sources are within a spectrum visible to a human eye.

6. The system of claim 1 , wherein there are at least three light sources and one is positioned to perpendicularly face the rear face of the optical redirection nodes, one is positioned to

perpendicularly face one of the side faces of the optical redirection nodes, and one is positioned to perpendicularly face the other side face of the optical redirection nodes.

7. The system of claim 3, wherein the at least two optical redirection nodes are coupled together using an optically clear adhesive.

8. The system of claim 1 , wherein there are at least three optical redirection nodes arranged in a row and coupled together using an optically clear adhesive.

9. The system of claim 1 , wherein one of the one or more light sources is coupled to one of the one or more collimating lenses and they are encased in a housing.

1 0. The system of claim 1 , wherein the one or more light sources are light emitting diodes.

1 1 . The system of claim 1 , wherein the one or more optical redirection nodes are cubic optical prisms.

1 2. The system of claim 1 , wherein the one or more optical redirection nodes are beam splitters.

1 3. The system of claim 9, wherein the system is further comprised of a base plate configured to receive one or more of the housings of the one of the one or more light sources coupled to the one of the one or more collimating lenses.

1 4. The system of claim 1 3, wherein the one or more optical redirection nodes are coupled to the base plate.

1 5. The system of claim 1 , wherein the one or more light sources are lasers.

1 6. The system of claim 1 3, wherein the housing is configured to be removably coupled to the base plate.

1 7. The system of claim 1 , wherein there are at least five light sources and one is positioned to perpendicularly face the rear face of the optical redirection nodes, one is positioned to perpendicularly face one of the side faces of the optical redirection nodes, one is positioned to perpendicularly face the other side face of the optical redirection nodes, one is positioned to perpendicularly face the top face of the optical redirection nodes, and one is positioned to perpendicularly face the bottom face of the optical redirection nodes.

1 8. A microscope system, the system comprising :

a multi-channel LED lighting array comprising:

one or more collimating lenses, each positioned between one of the one or more light sources and one of the one or more optical redirection nodes;

a microscope assembly comprising an objective, one or more magnifying lenses, an optical path, and a specimen stage.

1 9. The system of claim 1 8, wherein the objective is a digital display.

20. A microscope system, the system comprising :

a multi-channel LED lighting array comprising :

one or more cubic optical prisms, the cubic optical prisms comprised of a front face, a rear face, a top face, a bottom face, and two side faces;

three or more light emitting diodes, each positioned to face one of the front face, rear face, top face, or bottom face of one of the one or more cubic optical prisms; and

three or more collimating lenses, each positioned between one of the three or more light emitting diodes and one of the three or more cubic optical prisms;