I. Field of the Invention
The embodiments disclosed in this application generally relate to using a wavefront encoding apparatus to increase the depth of field in a short-wavelength microscope.
II. Background of the Invention
Microscopy and tomography of biological and other materials based upon short-wavelength radiation (i.e., soft x-rays, etc.) is a growing area of interest. There are currently, for example, a handful of soft x-ray microscopes available to researchers for biological work. Unfortunately they each are required to be located at a synchrotron facility. Obviously there are several disadvantages to this over a tabletop system. Researchers must schedule work well in advance, which precludes near real time research. Also, traveling to a synchrotron facility requires expense and time in travel and setup that decreases a researcher's productivity. Finally, since the facilities are shared, projects are ranked and reviewed, a process that can stifle some of the most innovative projects. In contrast a user owned x-ray microscope permits rapid turn around, high productivity, and the ability to run numerous experiments, even those that might be considered too high risk for a national facility.
When imaging samples using a short-wavelength microscope, both the lateral resolution and the depth of field of the microscope are important. For example, typical samples for soft x-ray microscopes are on the order of about 8 microns in diameter. Accordingly, to accurately image the sample through its entire depth, a depth of field of about 4 μm is required. The depth of field of a microscope is inversely related to the numerical aperture of the objective lens. Thus, for high power objective lenses, the numerical aperture will be much greater and the depth of field will generally be much smaller, which may lead to an insufficient depth of field. Conversely, if the depth of field is expanded in a short-wavelength microscope, too much of its magnification power or numerical aperture may be sacrificed.
Typically optical designers make extended depth of focus systems by stepping down the aperture until the desired depth of focus is realized. The two primary issues with doing this is that less light is collected by the objective lens (by the square of the diameter) and the resolution of the optical system is decreased due to low pass spatial filtering. Current short-wavelength objective zone plates (i.e., objective lens) are being manufactured with outer zone widths of as small as about 15 nm. As the outer zone width decreases the resolution of the short-wavelength microscope increases (linearly), however the depth of focus of the microscope decreases (by the square) of the outer zone width making high resolution single cell soft x-ray tomography (the real advantage of an short-wavelength microscope) very difficult if not impossible.
Apparatuses, methods, and systems for a short-wavelength microscope that can effectively expand its depth of field without sacrificing too much of its magnification power or numerical aperture are disclosed.
In one aspect, a lens assembly for enhancing the depth of field of a short-wavelength microscopic system is disclosed. The lens assembly includes an objective zone plate lens, an encoding lens, an imaging detector and a decoding component connected to the imaging detector. The objective zone plate lens is oriented to receive short-wavelength radiation that has passed through a sample in a microscopic system. The encoding lens is oriented to receive the short-wavelength radiation that has passed through the objective zone plate lens and encode the radiation to output an encoded short-wavelength radiation. The imaging detector is oriented to receive the encoded short-wavelength radiation and convert it to a digital signal which is subsequently decoded by the decoding component. The decoding applied by the decoding component effectively decodes the encoding applied to the short-wavelength radiation.
In another aspect, a short-wavelength microscopic device includes a laser device, a target, a condenser zone plate, a sample stage, an objective zone plate, an encoding lens, and an imaging detector connected to a decoding component. The laser device is configured to emit laser pulses which are received by the target. The target converts the laser pulses into short-wavelength radiation that is received by the condenser zone plate configured to form a diffraction pattern having a focal spot at where the sample stage is positioned. The sample stage is configured to mount a specimen sample. The objective zone plate is operable to receive the short-wavelength radiation that has passed through the specimen sample. As the encoding lens receives the short-wavelength radiation, it encodes the radiation to output an encoded short-wavelength radiation received by the imaging detector. The imaging detector is configured to convert the encoded short-wavelength radiation to a digital signal which is then decoded by the decoding component.
In still another aspect, a method for increasing the depth field in a short-wavelength microscopic device is disclosed, wherein the device includes: a condenser zone plate that is operable to receive short-wavelength radiation and form a diffraction pattern having a first order focal spot, a sample stage that can mount a specimen sample and can be operable to be positioned at the first order focal spot, and an objective zone plate lens operable to receive short-wavelength radiation that has passed through the specimen sample and focus the radiation onto an encoding element. The encoding element is configured to encode the short-wavelength radiation to output an encoded short-wavelength radiation. An imaging detector is positioned to receive the encoded short-wavelength radiation. The imaging detector transforms the encoded short-wavelength radiation into a digital signal. The digital signal is sent to the a decoding component. The decoding component reverses the encoding applied to the short-wavelength radiation.
These and other features, aspects, and embodiments of the invention are described below in the section entitled “Detailed Description.”
For a more complete understanding of the principles disclosure herein, and the advantages thereof, reference is now made to the following descriptions taken in conjunction with the accompanying drawings, in which:
An exemplary system for enhancing the depth of field in a short-wavelength microscope system 100 is depicted in
A condenser zone plate 115 captures some of the X-rays (or short-wavelength radiation) emitted by the target 110 and focuses those X-rays onto a focal spot, preferably on a sample stage 120. After the X-rays pass through the sample 120, they are captured by an objective lens 125, which preferably comprises another zone plate lens. It will be understood, however, that other types of lenses can be used depending on the requirements of the particular embodiment. After passing through the objective zone plate lens 125, the X-rays are passed to an imaging device 130, such as a CCD array.
The microscope system 100 has a depth of field 140, which is the zone of acceptable resolution of the imaged sample. The sample stage 120 can be moved between the condenser 115 and the objective 125 to align the specimen with the focal spot of the condenser zone plate lens 115. The depth of field 140 is determined by a variety of factors, including the wavelength of the illumination, the index of refraction of the medium surrounding the sample, the numerical aperture of the condenser and objective lenses (115, 125), and the magnification power of the objective lens 225. For many applications, it is desirable for the depth of field to be as large as possible, so that a larger range of features in the sample can be resolved in one image. As mentioned previously, however, the depth of field of high power objective lenses is very limited.
It has been discovered that obscuring one or more of the inner rings of the objective zone plate lens 125 will increase the depth of field 140 of microscope system 100. For example, an opaque plate 135 can be placed adjacent to the objective zone plate lens 125 so that one or more of the innermost rings of the objective zone plate array 125 are obscured. Another technique for obscuring the innermost rings in the objective zone plate lens 125 is to form the obscuring layer 135 directly on the objective zone plate 125 itself. This can be done by affixing an opaque plate 135 directly onto a pre-existing zone plate 125, or by fabricating a zone plate with one or more of the innermost rings being opaque rather than transparent. By obscuring the innermost rings of the objective zone plate lens 125, the image formed on the imager 130 will be formed only by the X-rays diffracted by the outer rings of the objective zone plate lens 125.
An alternative view of a microscope system 200 consistent with these principles is depicted in
The effects of the obscuring the innermost rings of an objective zone plate lens in a short-wavelength microscope are depicted in
In another embodiment, the tomographic capability (i.e., depth of field) can be extended and the imaging time for a short-wavelength microscope can be reduced by using wavefront encoding in the short-wavelength (i.e., soft x-ray regime). Wavefront encoding is a term used to describe the insertion of a special phase plate near the objective lens that extends the depth of focus of the optical imaging system by as much as a factor of 10 over a conventional imaging system. This can permit an increase in achievable resolution by a factor of three, yet retain the capability of short-wavelength tomography on, e.g., whole cells. In addition, enhanced depth of focus should relax the requirement that the illumination spectral bandwidth match the number of zones in the objective zone plate since the enhanced depth of focus allows a broadening of the radiation by the same eight to ten factor.
The short-wavelength radiation is directed at a condenser zone plate 115 which is positioned to capture the radiation and focus them onto a focal spot, which in this embodiment is the sample stage 120. After the short-wave radiation passes through the sample 120, it is captured by an objective lens 125. In one embodiment, the objective lens 125 is a zone plate type lens such as a Fresnel zone plate. It should be appreciated that the objective lens 125 can essentially be any type of lens as long as the objective lens 125 can be configured to optimally focus the short-wave radiation towards an intended target.
Continuing with
The basic principle behind wavefront coding imaging is to uniformly encode (i.e., blur) the short-wavelength radiation conveying the sample image from all planes to desensitize the image to depth of field 140 distortions (i.e., focus), capturing the image onto a digital imager designed for capturing the encoded radiation and converting the image to a digital signal, and decoding the digital signal to remove the effects (i.e., blurring) of the original encoding.
As depicted herein
In one embodiment, the encoding element 502 is a cubic phase plate lens that (i.e., encoding element 502) effectuates a phase shift encoding technique. The cubic phase plate lens should be manufactured on a thin x-ray transmissive membrane such as silicon nitride. The, e.g., nickel coating should be very thin, on the order of about 100 nanometers (nm) and should be processed to have a cubic functional form. The processing can be performed using standard lithographic techniques, possibly using an ion beam milling machine to impose the cubic functional form on the surface of the nickel. A surface profiling instrument, such as an Atomic Force Microscopy (AFM) instrument, can be used to make sure the nickel has the required shape. Examples of other suitable materials that the cubic phase plate lens can be fabricated from includes polymer-based substrates and zinc sulfide. However, it should be appreciated that the cubic phase plate lens can essentially be made out of any material as long as the lens can effectively encode short-wavelength radiation in accordance with the particular encoding technique being used.
The cubic phase plate (i.e., encoding element 502) phase shifts the short-wavelength radiation in accordance to an algorithmic step function. It should be understood that essentially any type of encoding element 504 and corresponding encoding technique can be employed to alter the short-wavelength radiation as long as the image conveyed by the short-wavelength radiation is “blurred” to produce an encoded image that is nearly independent of focus and the encoding technique can be later reversed by a decoding component 504.
Still with
After the encoded radiation is converted into a digital signal by the imaging detector 130, the signal is communicated to a decoding component 504 that is configured to reverse the encoding applied to the original non-encoded short-wavelength radiation. In one embodiment, decoding component 504 is a microprocessor or equivalent device that is configured to apply a logical algorithm (mathematical or otherwise) to the digital signal to reverse the encoding (i.e., decode the digital signal). In another embodiment, the decoding component 504 is a circuit that is configured to apply a logical algorithm to the digital signal to reverse the encoding. Following the decoding of the digital signal by the decoding component 504, the digital signal can be displayed on a high resolution display 506 such as a liquid crystal display (LCD) or equivalent device.
Although a few embodiments of the present invention have been described in detail herein, it should be understood, by those of ordinary skill, that the present invention may be embodied in many other specific forms without departing from the spirit or scope of the invention. Therefore, the present examples and embodiments are to be considered as illustrative and not restrictive, and the invention is not to be limited to the details provided therein, but may be modified and practiced within the scope of the appended claims.
This application claims the benefit of U.S. Provisional Application No. 60/701,842 filed Jul. 22, 2005. Additionally, this application is a continuation-in-part of prior application Ser. No. 11/161,880 filed Aug. 19, 2005 entitled “Method and Apparatus for Enhanced Depth of Field in X-Ray Microscopy Using Objective Zone Plate Obscuration.” The disclosure of each of the above-identified applications is incorporated herein by reference.
Number | Date | Country | |
---|---|---|---|
60701842 | Jul 2005 | US |
Number | Date | Country | |
---|---|---|---|
Parent | 11161880 | Aug 2005 | US |
Child | 11459605 | US |