Patent Application
20080144167
Embodiments of the invention relate generally to optical imaging systems and more specifically to reflective microscopes with aspheric objectives.
While examining biomedical specimens, it is often required to image dye stained objects or samples with various spectral outputs, such as ultraviolet, deep-blue or near-infrared. Dyes are usually used because various cell structures and objects are more easily observed with the eye once pigmented. Further, the dye molecules can be made to attach to specific sites within a specimen.
Typically, microscopes such as refractive microscopes are usually not configured to transmit ultraviolet or far infrared illumination (due to low glass absorption). In contrast, reflective microscopes have been used to capture images of the object in different spectral regions. However, such reflective microscopes typically have limited field of view for a given amount of resolution due to distortion, field curvature and other off axis aberrations. The limited field of view is a disadvantage when trying to image a large area of specimen or tissue. To overcome the disadvantage of a limited field of view, multiple smaller images of the object with higher levels of overlap are typically captured and then processed to form a single image. This process is typically slow and computationally intensive. The processed image, further, may have inaccuracies due to overlapping sections of the images.
In some instances, reflective microscopes have utilized reflective objectives with a transmissive field flattener between the objective and the sample to extend the field of view. However, such designs often have higher levels of chromatic aberration, and difficulty in transmitting the UV excitation illumination. The design of the reflective elements is strongly affected by the presence of the refractive field flattener in front of the reflective elements, so changing the materials and form of the field flattener to allow use over multiple spectral sources is not practical.
In typical microscope designs, both reflective and refractive, the lenses are constructed in such a way as to allow the simultaneous viewing of specimens by eye and by digital sensor. Further the optical path is arranged in such a way to allow the addition of various optical components (e.g. filter, polarizers, beam splitters). The impact of this approach is often a loss of collected light from the specimen. The loss of light, in turn, leads to a reduction in image signal-to-noise or contrast-to-noise that must be compensated with more illumination power and or longer exposure time on the image sensor. This is a disadvantage in high-speed imaging applications.
Typical microscope objectives do not typically allow for low-loss transmission of ultraviolet and visible light. In certain types of epi-fluorescent imaging, the objective lens must simultaneously create a high-quality visible light image and transmit a light source for the excitation of a fluorescent dye or particle. Refractive microscope objectives are not easily suited to this task if the desired excitation is in the UV spectrum, e.g. 355 nm as used to excite quantum dots particles.
Therefore, there is a need for a microscope with a wide field of view and which is adapted for rapidly imaging an object over multiple spectral ranges at a high resolution.
Briefly, in accordance with one embodiment of the invention, an optical imaging apparatus is provided. The optical imaging apparatus includes an aspheric objective configured to receive optical radiation from an object. The aspheric objective comprises a first reflective aspheric mirror. A second reflective aspheric mirror is optically coupled to the first reflective aspheric mirror such that optical radiation received from the object is reflected by the first reflective aspheric mirror to the second reflective aspheric mirror. The first reflective aspheric mirror and the second reflective aspheric mirror are centered on a single, optical axis. The optical imaging apparatus further includes an optical sensing device disposed adjacent to a non-reflective side of the first aspheric mirror and positioned to receive the optical radiation reflected by the second reflective aspheric mirror and configured to render digitized images of reflected optical radiation representative of the object.
In another embodiment, an optical imaging method is provided. The method comprises receiving optical radiation incident upon a first reflective aspheric mirror and reflecting the optical radiation from the first reflective aspheric mirror to a second reflective aspheric mirror. The first reflective aspheric mirror and the second reflective aspheric mirror are centered on a single, optical axis. The method further comprises reflecting the optical radiation from the second reflective aspheric mirror to an imaging device and rendering digitized images of the optical radiation reflected by the second reflective aspheric mirror.
In another embodiment, a reflective microscope for wide field of view imaging is provided. The reflective microscope comprises a holder for placing an object to be imaged and an aspheric objective configured to receive optical radiation from the object. The aspheric objective comprises a first reflective aspheric mirror and a second reflective aspheric mirror optically coupled to the first reflective aspheric mirror such that optical radiation received from the object is reflected by the first reflective aspheric mirror to the second reflective aspheric mirror. The first reflective aspheric mirror and the second reflective aspheric mirror are centered on a single axis. The reflective microscope further comprises an optical sensing device disposed adjacent to a non-reflective side of the first aspheric mirror and positioned to receive the optical radiation reflected by the second reflective aspheric mirror and configured to render digitized images of reflected optical radiation representative of the object. The reflective microscope also includes a field flattener disposed between the second reflective aspheric mirror and the sensing device such that the optical radiation reflected by the second reflective aspheric mirror is received by the optical sensing device after being transmitted through the field flattening lens and an image processor to receive the digitized images and configured to generate an image of the object.
These and other features, aspects, and advantages of the present invention will become better understood when the following detailed description is read with reference to the accompanying drawings in which like characters represent like parts throughout the drawings, wherein:
The aspheric objective 12 comprises a first reflective aspheric mirror 22 and a second reflective aspheric mirror 24. The first reflective aspheric mirror 22 comprises a reflective surface 26 and non-reflective surface 28. Similarly, the second reflective aspheric mirror 24 comprises a reflective surface 30 and a non-reflective surface 32. The reflective surface 26 of first reflective aspheric mirror 22 faces the reflective surface 30 of the second reflective aspheric mirror 24.
The first reflective aspheric mirror 22 and the second reflective aspheric mirror 24 are centered on a single optical axis. In one embodiment, the mirrors are diamond turned to provide a surface finish to a precision of about 2-3 nm. In one embodiment, the outside diameters of the two aspheric mirrors are cut on the same machining fixture as the optical surfaces so as to facilitate centering of their respective optical axes. In a specific embodiment, a radius of the first reflective aspheric mirror 22 is greater than the radius of the second reflective aspheric mirror 24. In a specific embodiment, the radius of the first reflective aspheric mirror ranges between 1.9 to 2.2 times the radius of the second reflective aspheric mirror. The use of reflective aspheric mirrors provides a higher level of aberration correction over a wider field of view. In one embodiment, field of view is wider by 40-80% compared to existing refractive microscope objectives.
The surfaces of the first and second reflective aspheric mirrors are characterized by curvatures.
In one embodiment, the curvature of an aspherical surface 36 may be defined by an nth order polynomial. In one embodiment, the curvature of the first and second reflective aspheric surfaces is characterized by a fourth or higher order polynomial. In one embodiment, the curvature of the first and second reflective aspheric surfaces is characterized by a tenth order polynomial with a conic constant. A tenth order polynomial representing the curvature of the first and second reflective aspheric surfaces may be represented by the following equation:
Continuing with
Field flattener 14 is disposed between the non-reflective side 28 of the first reflective aspheric mirror 22 and the optical sensing device 16. In the illustrated embodiment, the field flattener 14 is a refractive field flattener and is disposed such that the optical radiation reflected by the second reflective aspheric mirror 24 is received by the optical sensing device 16 through the field flattener. The field flattener is designed in accordance with the optical sensing device being used. As the field flattener is an achromatic doublet, which substantially reduces variation in focal length with wavelength. The aberrations in the visible range are reduced by bringing the red light and blue light focus to the same plane, whereas normally the best focus for the red, green and blue are linearly displaced along the optical axis. In one embodiment, the distortion is about 0.5%.
In one embodiment, the field flattener 14 may comprise a concave lens and a convex lens coupled together. In one embodiment, the concave lens has a high Abbe number and low dispersion. The convex lens, on the other hand has a low Abbe number and high dispersion. As is well known, the Abbe number of a transparent material is a measure of the material's dispersion or a measure of variation of refractive index with wavelength. For one embodiment in the visible wavelength range, the lenses are formed with Schott BK7 (refractive index 1.517, Abbe number 64.2 (low dispersion negative element)), Schott F2 (refractive index 1.620, Abbe number 36.4 (high dispersion positive element)) optical glass. For the infrared wavelength range the relevant materials include Silica and Germanium. For the ultraviolet wavelength range the relevant materials include fused silica and calcium fluoride. In other embodiments, a reflective field flattener may be used in place of a refractive field flattener. The reflective field flattener can operate over the ultraviolet wavelength, visible wavelength and the infrared wavelength.
In the illustrated method, optical radiation from an object is received by a first reflective aspheric mirror such as reflective aspheric mirror 26 at block 42. The optical radiation may be received by either transmissively illuminating the object or by reflectively illuminating the object. An imaging apparatus using a bottom-illuminated object is defined as a tranmissive illumination system. An imaging apparatus using a top illuminated object is defined as a reflected illumination. At block 44, the optical radiation from the first reflective aspheric mirror is reflected to a second reflective aspheric mirror such as reflective aspheric mirror 24.
At block 46, the optical radiation is reflected from the second reflective aspheric surface and is transmitted to an imaging device such as optical sensing device 16. In a further embodiment, the optical radiation reflected by the second reflective aspheric mirror is transformed into a flatter image with the field flattener lens before being transmitted to the image sensor. In one embodiment, the optical radiation is flattened using an achromatic field flattener such as field flattener 14.
Finally, in block 48, digitized images of the object are rendered using the optical radiation reflected by the second reflective aspheric mirror and transmitted to the image sensor. In one embodiment, the rendering is performed using an optical sensing device.
In one embodiment, the optical radiation is received by transmissively illuminating the object as shown in
In another embodiment, the optical radiation is received by a dark field reflected illumination (or epi-illumination) as shown in
The reflective microscope 62 comprises aspheric objective 64 and image side tube 66. Each component is described in further detail below. A holder (not shown) is provided for retaining an object for microscopic imaging. The holder is typically disposed adjacent to objective 64. The distance between the holder and the objective is generally referred to as working distance. In one embodiment, the working distance of the reflective microscope is greater than 5 millimeters. In a more specific embodiment, is the working distance is about 55 millimeters.
The aspheric objective 64 is configured to receive optical radiation from the object to be imaged. The aspheric objective 64 comprises a first reflective aspheric mirror 68 and a second reflective aspheric mirror 70. The first and second reflective aspheric mirrors are implemented similar to the techniques described in
Image side tube 66 houses the field flattener 72 and the optical sensing device 74. The field flattener comprises a reflective field flattener or a refractive field flattener. The reflective microscope is designed such that a user may place the desired type of field flattener within the image side tube based on the object being examined.
In one embodiment, the length of the image side tube is about 319 mm. In another embodiment, the distance between the field flattener and the objective is about 231 mm and the distance between the field flattener and the optical sensing device is about 83 mm. The image side tube may further include an image processor (not shown) to receive the digitized images and configured to generate an image of the object, which can be viewed on a display device (not shown).
The embodiments described above have many advantages including the ability to operate in multiple spectral ranges by using the aspheric reflective mirrors. By using the aspheric reflective mirrors, it is possible to eliminate the need for a large number of refractive lenses in a microscope. Thus, by changing the illumination configuration, the microscope may be used for bright field imaging, dark field imaging and fluorescence imaging. By using a field flattener in conjunction with the aspheric reflective surfaces, high resolution images with a wider field of view can be obtained.
While only certain features of the invention have been illustrated and described herein, many modifications and changes will occur to those skilled in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.
