Confocal scanning microscopy (CSM) needs a scanning mechanism because only one point information of specimen can
be obtained. Therefore the speed of the confocal scanning microscopy is limited by the speed of the scanning tool. To
overcome this limitation from scanning tool we propose another scanning mechanism. We make three optical probes in
the specimen under confocal condition of each point. Three optical probes are moved by beam scanning mechanism with
shared resonant scanning mirror (RM) and galvanometer driven mirror (GM). As each optical probe scan allocated
region of the specimen, information from three points is obtained simultaneously and image acquisition time is reduced.
Therefore confocal scanning microscopy with multiple optical probes is expected to have three times faster speed of the
image acquisition than conventional one. And as another use, multiple optical probes to which different light wavelength
is applied can scan whole same region respectively. It helps to obtain better contrast image in case of specimens having
different optical characteristics for specific light wavelength. In conclusion confocal scanning microscopy with multiple
optical probes is useful technique for views of image acquisition speed and image quality.
A new technique for improving the axial resolution of confocal microscope is proposed. Based on the interference
between two different frequency beams, which are separated axially, a frequency domain field confined focal spot is
generated. The effective region made by the interference makes the point-spread function (PSF) of confocal microscope
sharper. The three-dimensional imaging equations are derived. The intensity distribution of frequency domain field
confined focal spot is proportional to the absolute value of the product of two fields. Three-dimensional intensity pointspread
function (IPSF) is calculated numerically. The farther two beams are separated axially, the sharper IPSF is
obtained. The numerical results show that the full width half maximum (FWHM) of the IPSF is improved by factor of
1.78 maintaining the strength of side lobe at 0.5 relative to main lobe. Also simulations for two-point resolution show the
same improvement in the axial resolution.
KEYWORDS: Signal to noise ratio, 3D modeling, Confocal microscopy, Microscopy, 3D image reconstruction, 3D image processing, 3D metrology, Image processing, Time metrology, Actuators
Confocal Scanning Microscopy (CSM) is very useful to reconstruct 3D image of Bio-cells and the objects that have specification shape in higher axial and lateral resolution and widely used as measurement instrument. A 3D reconstruction is used to visualize confocal images and consists of following processes. The First process is to get 3D data by collecting a series of images at regular focus intervals (Optical Sectioning). The Second process is to fit a curve to a series of 3D data points each pixel. The Third process is to search height information that has maximum value from curve-fitting. However, because of various systematic errors (NOISE) occurred when collecting the information of images through Optical Sectioning and large peak deviation occurred from curve-fitting error, high quality 3D reconstruction is not expected. Also, it takes much time to 3d Reconstruction by using many 3D data in order to acquire high quality and much cost to improve signal-to-noise (SNR) using a higher power laser. So, we are going to define SNR, peak deviation and the order of curve-fitting as important factors and simulate the relation between the factors in order to find a optimum condition for high quality 3D reconstruction in Confoal Scanning Microscopy. If we use optimum condition obtained by this simulation, using a suitable SNR and the suitable number of data and the suitable n-th order curve-fitting, small peak deviation is expected and then, 3D reconstruction of little better quality is expected. Also, it is expected to save.
The errors can cause the serious loss of the performance of a precision machine system. In this paper, we propose the method of allocating the alignment tolerances of the components and apply this method to Confocal Scanning Microscopy (CSM) to get the optimal tolerances.
CSM uses confocal aperture, which blocks the out-of-focus information. Thus, it provides images with superior resolution and has unique property of optical sectioning. Recently, due to these properties, it has been widely used for measurement in biological field, medical science, material science and semiconductor industry.
In general, tight tolerances are required to maintain the performance of a system, but a high cost of manufacturing and assembling is required to preserve the tight tolerances. The purpose of allocating the optimal tolerances is minimizing the cost while keeping the performance of the system. In the optimal problem, we set the performance requirements as constraints and maximized the tolerances.
The Monte Carlo Method, a statistical simulation method, is used in tolerance analysis. Alignment tolerances of optical components of the confocal scanning microscopy are optimized, to minimize the cost and to maintain the observation performance of the microscopy. We can also apply this method to the other precision machine system.
We describe the design and the implementation of reflection confocal scanning microscopy (CSM) using an acousto-optical deflector (AOD) for the fast horizontal scan and a galvanometer mirror (GM) for the slow vertical scan. In the beam scanning system it is important to maintain the lateral and the axial performance during scanning operation. We propose a simple method to design a scanning system using the finite ray tracing and the diffraction theory. We define a
cost function which contains the effect of aberrations on the performance of microscopy. We construct the designed system and evaluate its performance. The OSLO simulation shows that the performances of CSM are not changed with deflection angle. So we conclude that the beam scanning system is properly designed. In addition, we propose an image formation method and show images obtained with the system.
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