Patent Application
20090161210
This application claims priority of No. 096149047 filed in Taiwan R.O.C. on Dec. 20, 2007 under 35 USC 119, the entire content of which is hereby incorporated by reference.
1. Field of Invention
The invention relates in general to a microscopy system, and more particularly to a microscopy system having a revolvable stage.
2. Related Art
Confocal laser scanning microscopy (CLSM or LSCM) is a valuable tool for obtaining high resolution images and 3-D reconstructions by using a spatial pinhole to eliminate out-of-focus light or flare. This technology permits one to obtain images of various Z-axis planes (Z-stacks) of the sample. The detected light originating from an illuminated volume element within the specimen represents one pixel in the resulting image. As the laser scans over the plane of interest, a whole image is obtained pixel by pixel and line by line. The beam is scanned across the sample in the horizontal plane using one or more (servo-controlled) oscillating mirrors. Information can be collected from different focal planes by raising or lowering the microscope stage. The computer can calculate and then generate a three-dimensional picture of the specimen by assembling a stack of these two-dimensional images from successive focal planes.
However, the Z-axis direction in the stacked 3D image has a much poor resolution (e.g., about 1.2 μm/slice) than in the X-axis and Y-axis directions (about 0.15 μm/pixel) under the limitation of the dimension of the pinhole and other mechanical or physical properties. A poor resolved Z-axis direction hampers the spatial reliability of the high resolution neural network images reconstructed, especially when comparison of two different samples is necessary. One of the inventors, Ann-Shyn Chiang, has disclosed an aqueous tissue clearing solution in U.S. Pat. No. 6,472,216 B1. In the '216 patent, the depth of observation may reach the level of micrometers. In the current developing method, fluorescent molecules are attached to or combined with the biological tissue. Thus, making the tissue become transparent is a key point for the break-through of the depth of observation, and the way of solving the bottleneck of the Z-axis resolution is greatly needed.
It is therefore an object of the invention to provide a microscopy system with a revolvable stage for revolving a sample and holding the sample in a saturated condition so that the resolution of 3D image sensing can be increased.
The invention achieves the above-identified object by providing a microscopy system including an objective lens and a stage for holding a sample. The objective lens focuses incident light coming from one side of the objective lens to the sample disposed on the other side of the objective lens, and focuses an optical signal emitted from the sample to a photosensor disposed on the one side of the objective lens. The stage supports the sample and is configured to be revolvable about an axis, which is substantially perpendicular to an extending direction from the sample to the objective lens.
Further scope of the applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.
The present invention will become more fully understood from the detailed description given hereinbelow and the accompanying drawings which are given by way of illustration only, and thus are not limitative of the present invention.
The present invention will be apparent from the following detailed description, which proceeds with reference to the accompanying drawings, wherein the same references relate to the same elements.
The present inventors have found that the sample may be revolved by a specific angle about an X-axis or a Y-axis. Then, the image synthesis may be performed by way of image processing in order to solve the problem of the too-low resolution in the Z-axis direction. In order to achieve this effect, a stage for supporting and holding the sample has to be configured to be revolvable. It is to be noted that the term “revolvable” covers the rotatable ranges from 0 to 360 degrees, and this rotation may be out of the plane of the microscope platen. That is, the axis of rotation is not perpendicular to the plane of the microscope platen. The detailed structure of the microscopy system of the invention will be described in the following.
The objective lens 10 focuses incident light L1, coming from one side of the objective lens 10, to the sample 12 disposed on the other side of the objective lens 10, and focuses an optical signal, emitted from the sample 12, to a photosensor 5 disposed on the one side of the objective lens 10. The stage 14 supports the sample 12 and is configured to be revolvable about an axis 18, which is substantially perpendicular to an extending direction 16 from the sample 12 to the objective lens 10, as shown in
When being applied to the CLSM, the microscopy system may further include a light source 1, a light input aperture 2, a beam splitter 3, a light output aperture 4 and the photosensor 5. For example, the light source 1, such as a laser light source, outputs the incident light L1 to the sample 12 sequentially through the light input aperture 2, the beam splitter 3 and the objective lens 10 so that reflected light L2 is generated. The reflected light L2 passes through the objective lens 10 and is reflected, by the beam splitter 3, to the photosensor 5 through the light output aperture 4.
In one example, the stage 14 may also be configured to be movable along the extending direction 16. Therefore, the photosensor 5 may sense the sample 12 disposed on a focus plane FP so that the stage 14 can be moved along the extending direction 16, the sample 12 can be moved along the extending direction 16, and various images at various depths of the sample 12 may be located on the focus plane FP.
The microscopy system with the revolvable stage according to the invention makes the sample be revolvable. In addition, many layers, which are stacked in the Z-axis direction and are gathered at different revolving angles, such as 0 and 90 degrees, can be integrated. So, it is possible to reconstruct a three-dimensional image having the high resolution at three primary axes, and thus to implement other diversified image sensing functions.
While the invention has been described by way of examples and in terms of preferred embodiments, it is to be understood that the invention is not limited thereto. To the contrary, it is intended to cover various modifications. Therefore, the scope of the appended claims should be accorded the broadest interpretation so as to encompass all such modifications.
| Number | Date | Country | Kind |
|---|---|---|---|
| 096149047 | Dec 2007 | TW | national |
