Tandem Scanning Confocal microscopy (TSCM) is ideal for in vivo corneal imaging, but currently requires time-consuming off-line imageprocessing. To overcome this limitation, we have developed a novel PC-based TSCM sy...
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ISBN:
(纸本)0780356756
Tandem Scanning Confocal microscopy (TSCM) is ideal for in vivo corneal imaging, but currently requires time-consuming off-line imageprocessing. To overcome this limitation, we have developed a novel PC-based TSCM system. The system is the first to provide interactive 3-D on-line imageacquisition and analysis of in vivo confocal images.
Modem research in molecular, cellular, and developmental biology requires the precise measurement of cellular or subcellular activity in two- or three-dimensions. Many available fluorescence microscope techniques are ...
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ISBN:
(纸本)0819430749
Modem research in molecular, cellular, and developmental biology requires the precise measurement of cellular or subcellular activity in two- or three-dimensions. Many available fluorescence microscope techniques are yielding limited detail about the organization and dynamics of complex cellular structures. But the multiphoton (two- or more photon) excitation fluorescence imaging microscopy (MEFIM) system proposed here will provide a unique, state-of-the-art opportunity to integrate advances in optical microscopy, low-light video detection, and two- or three-dimensionalimage analysis in measuring the fluorescent signals from living cells. The MEFIM system uses longer excitation wavelength, which increases the penetration of the excitation of the sample, yet essentially reduces photobleaching and autofluorescence. But, more importantly, the infrared light excitation in the MEFIM techniques allows maintaining cell viability for longer periods of time and thus acquisition of more images. In our current work, we integrated the MEFIM system with one-photon laser scanning confocal microscope coupled to a verdi pumped tunable femtosecond pulsed ti-sapphire laser. Appropriate wavelength was tuned to excite the fluorescently labeled specimens and MEFIM images were acquired from living biological specimens at different optical sections. The importance of the MEFIM imageacquisition and processing for biomedical sciences are discussed.
Tandem Scanning Confocal microscopy (TSCM) is ideal for in vivo corneal imaging, but currently requires time-consuming off-line imageprocessing. To overcome this limitation, we have developed a novel PC-based TSCM sy...
详细信息
Tandem Scanning Confocal microscopy (TSCM) is ideal for in vivo corneal imaging, but currently requires time-consuming off-line imageprocessing. To overcome this limitation, we have developed a novel PC-based TSCM system. The system is the first to provide interactive 3-D online imageacquisition and analysis of in vivo confocal images.
An analysis is presented of the image formation in widefield fluorescence microscopy using standard light. The region of support of the resulting optical transfer function is discussed.
ISBN:
(纸本)0819427004
An analysis is presented of the image formation in widefield fluorescence microscopy using standard light. The region of support of the resulting optical transfer function is discussed.
This paper describes a method for the improvement of biological images acquired using a Transmission Electronic Microscope (TEM). Several techniques are presented that deal with noise reduction, artifact removal and n...
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ISBN:
(纸本)0819427004
This paper describes a method for the improvement of biological images acquired using a Transmission Electronic Microscope (TEM). Several techniques are presented that deal with noise reduction, artifact removal and non-uniform illumination correction. Experimental results are shown.
We describe a method of obtaining optically sectioned fluorescence images in a widefield conventional microscope by interfering two beams on an object so as to illuminate it with a single spatial frequency fringe patt...
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ISBN:
(纸本)0819427004
We describe a method of obtaining optically sectioned fluorescence images in a widefield conventional microscope by interfering two beams on an object so as to illuminate it with a single spatial frequency fringe pattern. images taken at three spatial positions of the fringe pattern are processed in real time to produce optically sectioned images which are substantially similar to those obtained with confocal microscopes.
In this work we discuss the role of polarisation in confocal microscopy. The effect of the optical system on light polarisation is analysed in the absence of a specimen. We also present numerical results on confocal m...
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ISBN:
(纸本)0819427004
In this work we discuss the role of polarisation in confocal microscopy. The effect of the optical system on light polarisation is analysed in the absence of a specimen. We also present numerical results on confocal microscopes imaging small dielectric scatterers and suggest an optimum pinhole size for crossed linear polars. Finally, imaging of large spherical scatterers and contrast optimisation of the optical system is discussed.
We introduce an optical means of reconstructing the topology of rough objects observed under the microscope or at a relatively high magnification. First, 2D images were obtained under the conventional optical microsco...
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ISBN:
(纸本)0819427004
We introduce an optical means of reconstructing the topology of rough objects observed under the microscope or at a relatively high magnification. First, 2D images were obtained under the conventional optical microscope or under the macroscopic imaging system, focused on differing height of the sample. The depth information was extracted by sensing the focus of each pixel of the conventional image slice. Each sample height associated with the best "focus figure" for each pixel was marked and were later utilized to generate the three dimensional coordinate map.
imageacquisition at high magnification is inevitably correlated with a limited view over the entire tissue section. To overcome this limitation we designed software for multiple image-stack acquisition (3D-MISA) in c...
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ISBN:
(纸本)0819427004
imageacquisition at high magnification is inevitably correlated with a limited view over the entire tissue section. To overcome this limitation we designed software for multiple image-stack acquisition (3D-MISA) in confocal laser scanning microscopy (CLSM). The system consists of a 4 channel Leica CLSM equipped with a high resolution z-scanning stage (Leica Lasertechnik) mounted on a xy-motorized stage (Marzhauser). The 3D-MISA software is implemented into the microscope scanning software (Scanware, Leica Lasertechnik) and uses the microscope settings (magnification, zoom) for the movements of the xy-stage. It allows storage and recall of 70 xyz-positions and the automatic 3D-scanning of image arrays between selected xyz-coordinates. The number of images within one array is limited only by the amount of disk space or memory available. Although for most applications the accuracy of the xy-scanning stage is sufficient for a precise alignment of tiled views, the software provides the possibility of an adjustable overlap between two image stacks by shifting the moving steps of the xy-scanning stage. After scanning a tiled image gallery of the extended focus-images of each channel will be displayed on a graphic monitor. In addition, a tiled image gallery of individual focal planes can be created. In summary, the 3D-MISA allows 3D-imageacquisition of coherent regions in combination with high resolution of single images.
Imaging thick specimens in 3D transmission confocal modes presents two key problems. The first problem is variable aberrations introduced by changes in refractive index. The second problem is revealed when visualising...
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ISBN:
(纸本)0819427004
Imaging thick specimens in 3D transmission confocal modes presents two key problems. The first problem is variable aberrations introduced by changes in refractive index. The second problem is revealed when visualising acquired data, where thick 3D datasets are difficult to interpret. In this paper we present our emerging solutions to these problems. Aberrations can be classified as simple tip-tilt deflection of the beam, or more complicated higher order aberrations. We discuss our results which demonstrate successful on-the-fly detection and correction for tip-tilt For detecting higher order aberrations, we have chosen to investigate the wavefront curvature sensing technique. The second problem of rendering thick 3D datasets can be solved by extracting features of interest from the background. Simple intensity threshholding is not sufficient for complex biological specimens, and imageprocessing in only two dimensions neglects any three dimensional structure. Use of Kohonen's self-organising map neural network in 3D results in clear segmentation of features for sample chromosome specimens.
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