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This invention relates to electro-optical systems and particularly to illumination systems for microscopy using a plurality of selectable sources.
Fluorescence microscopy is a fundamental tool in cellular biology. It is used by researchers to monitor cellular function, to investigate gene expression, and to evaluate new drug candidates among an ever-growing collection of applications. In general, fluorescence microscopy uses a bright light source with a predetermined wavelength to excite a fluorophore (chemical label) which then emits light at a wavelength different from the excitation. Typically, as part of an illumination system, excitation colors are conditioned or otherwise collected, shaped, and modified by optical components to provide efficient and even illumination at the image plane of the microscope (i.e. location of the fluorophore). It is well known that an illumination system that has a large number of bright selectable wavelengths is a valuable tool for cellular research.
The prior art includes a number of systems attempting to address this need using a variety of arrangements and excitation sources. Typical prior art systems include filtering a broad-spectrum light source (e.g. white light) to produce a desired excitation color. These systems are typically characterized by a large size, an undesirable amount of waste heat, and expensive, short-lived light sources. Other prior art systems optically mix specific colored sources using dichroic beamsplitters. These systems have been limited to relatively few sources by the complexity and cost of each additional dichroic element. Still other systems have attempted to circumvent the need for complex optics by mechanically switching between a number of specific colored sources. To date, these systems have been characterized by mechanical enclosures and mechanical mountings that significantly limit the number and variety of sources, as well as the speed of mechanical transitions between sources.
It is the purpose of the present invention to provide a novel system that addresses problems of the prior solutions. In addition, the present invention enables lower cost, higher efficiency microscopy and the potential for more rapid, economical cellular research. Further objects and advantages will become apparent from the detailed descriptions that follow.
The present invention is a novel electro-optical system providing controllable alignment of any of a plurality of electro-optical components mounted to a circuit board with an optical axis.
Its preferred embodiment provides selectable illumination wavelengths and selectable conditioning optics for microscopy. By exploiting the mechanical, electrical, and thermal properties of a circuit board as well as its efficient manufacture, the present invention provides a high density of wavelength options, low heat generation and low cost.
Using a circular arrangement of surface mounted light sources and directly coupling this arrangement to a controlled motor provides a fast switching, multi-source illumination system. The circular arrangement aligns a light source with a primary optical axis and can be arranged to align a second light source with a secondary optical axis. The low inertia of the circuit board assembly allows the motor to switch rapidly between different sources within the circular arrangement. The high density circular arrangement of light sources provides options for multiply redundant wavelengths, close spatial groupings of application specific wavelengths, and multiple source types including both light emitting diodes and laser diodes.
Electronics manufacturing technologies provide useful arrangements of circuit board features, control circuitry, and light sources. Additional electrical, thermal, and mechanical features can be integrated directly into the circuit board including temperature and light sensing components, clusters of through-holes for thermal management and flexible, cantilevered substrate regions providing a unique combination of structures.
The preferred embodiment of the present invention is an illumination system for a microscope. It generally comprises a light source assembly constructed from a circuit board with a geometry and pattern of light emitting components convenient for rotational positioning and alignment with multiple optical axes. An additional embodiment includes a selectable sensor array comprising surface mounted sensors with varying sensitivities to radiation. An arrangement combining the sourcing and the sensing of light provides another embodiment.
In addition, each location is characterized with a pattern of holes 35 drilled through the circuit board. These holes are tinned in a manner familiar in circuit board manufacture. In this case, the holes act as a heat sink for each LED component. Each mounted LED component is in thermal contact with a cluster of holes.
A control circuit, schematically shown as a single square region 36, but typically a collection of components, is mounted directly to the circuit board 30 and interfaces with a control computer (not shown). The interface is established via a cable connected using the connector 32. In operation, timing and command signals are directed via the control circuit out to one or more LED components using circuit traces on the circuit board. A portion of the circuit traces are shown as item 37.
In the embodiment of
In typical usage, some colors are more popular with microscopists. For example, users of fluorescent microscopy often allocate a 405 nm wavelength (violet) to mark the nucleus of cells. In this case, violet sources can be placed in multiple locations around the rotary platform. Thus, a sequence of colors including violet can strategically use the nearest violet source and reduce transition times between violet and non-violet sources. Furthermore, multiple instances of a given color provides redundancy and an improvement in mean-time-to-failure. Similarly, multiple wavelengths can be grouped in adjacent mount positions corresponding to popular multi-color (multi-plexed) assays further minimizing the required motion for a specific measurement.
The circuit board construction provides a number of alternative embodiments mixing a high density arrangement of mixed light sources, circuitry, and sensors.
Several sensors are included in this embodiment as items 42 and 43. Sensors can be mounted immediately adjacent to source components and control circuitry providing, cost effective, efficient, and reliable construction. Sensors and sources are directly mounted to the mechanical substrate without need for additional mounting means or additional cabling. For example, a photodiode can be used to detect a portion of the emitted light from a source and provide feedback for the control circuit. Light output intensity feedback allows consistent output intensity control and can be used as a detection means to locate failed or degrading light sources. Similarly, a temperature sensor mounted adjacent to one or more sources can monitor thermal conditions and detect unsafe operating conditions.
A specific embodiment and application of both active and passive electro-optical components is shown in
Additional Circuit Board Features
The mechanical, electrical, and thermal role played by the circuit board 30 enables further novel features demonstrated in the embodiment of
The circuit board is often constructed of a multi-layer fiberglass composite or ceramic and is both lightweight and strong; ideal for high speed rotary positioning applications.
The preferred embodiment additionally mounts the light source assembly as shown in
In operation, power and position control signals are directed through controller 82 to the stepper motor 81 and produce controlled rotation of the motor's shaft. Consequently, the backing disk, and the substrate attached to it, rotate an identical angular displacement. The controlled rotation positions a predetermined LED component in line with the primary optical axis 65 or a secondary axis. At substantially the same time, the optical encoder position is measured to confirm the angular position of the circuit board. After positioning, illumination commands (for example, current amplitude and current duration) are sent to aligned LED components.
In an additional embodiment shown in
It is known that laser illumination as a light source in microscopy is generally characterized by speckle in the image caused by structures in the sample that scatter the incident light which then produces a light and dark pattern of interference. This effect introduces spurious contrast and confounds image analysis. A diffusing component can be arranged to interact with a laser source 40 on a single circuit board assembly as shown in
A Light Source Assembly with Multiple Axes and Light Conditioning
In operation, a selected LED component 31b is positioned along the primary optical axis 65 by controlled rotation of mounted circuit board 30. A command signal (not shown) turns on LED 31b. The collection lens 91 (a conditioning optic) efficiently directs emitted light along the primary axis 65. All mounted LEDs can be aligned with at least one secondary axis. The mechanical spacing of mounts 92 and 92b allow alignment of axes simultaneously with several LEDs. For example, mount position 92b aligns with LED 31c and in this case mounted fiber 93. A command signal (not shown) turns on LED 31c independently of other LEDs.
An additional embodiment is shown in
Similarly,
Many other orientations of the rotatably mounted circuit board that still allow alignment of a collection of arranged optical components are within the scope of the present invention.
The Present Invention Can Stack Boards to Increase mounting area, bring additional heat sinking, peltier cooling, or opto-mechanical options into proximity with a primary platform. These stacked boards can operate similar to daughter boards familiar to the art of printed circuit board assemblies. In addition, several independently rotatable circuit board assemblies can be placed adjacent to one another and provide additional capabilities as a combined system.
An additional embodiment of a circuit board assembly is shown in
In operation, the circuit board 30′ is rotated to align a selected optical component with a selected LED source of circuit board 30. Thus, components of 30′ provide selectable conditioning to light sources mounted to 30. For example, an LED component 31e on circuit board 30 is aligned to the primary optical axis 65. In addition, a lens mounted to circuit board 30′ is aligned to the primary optical axis and provides predetermined conditioning to light emitted from 31 e. The conditioned light passes through the aperture 62. In another example, clear aperture 133, shown as a rectangle, is intended to act as an illumination mask. Several methods can be used to construct the rectangular mask including machining directly through the circuit board substrate, laminating a thin foil with a patterned mask (aperture) onto the circuit board, mounting a photolithographically patterned target onto the circuit board (e.g. chrome on glass).
In operation, this mask can be projected onto the focal plane of the microscope. The resulting projected pattern of light is a rectangle constructed to match the light sensitive area of a CCD camera. In this way, light illuminates only intended regions of the target that are viewed by the camera. This is advantageous when using fluorescent targets whose fluorophore can have a reduced useful lifetime when subjected to unintended light. Additional projected patterns for various structured illumination techniques can be added to circuit board 30′ such as a fine array of stripes, a grid of lines, or an array of pin holes.
In another example, a low angle diffusing component mounted on circuit board 30′ is arranged as a conditioning optic for a mounted laser diode 40 on circuit board 30. In operation, the position of the diffuser, perpendicular to the optical axis, is varied by controlled rotations of the circuit board 30′ to impart changes to the laser speckle and improve measurements.
The circuit board 30′ can provide a high density of illumination masks and conditioning components directly adjacent to control electronics and sensors. The sequence of two circuit board assemblies shown in
Additional alternative designs and assemblies are within the scope of this disclosure and although several are described they are not intended to define the scope of the invention or to be otherwise limiting.
This application claims the benefit of provisional patent application Ser. No. 61/797,413, filed Dec. 6, 2012.
Number | Date | Country | |
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61797413 | Dec 2012 | US |