Image Acquisitionand Proces

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Image Processing 99 As examples of our developments, we introduced program components for acquisition (FastRecord module) and analysis (FastAnalysis module) of image data obtained in the course of our brain research using laser microscopy. The entire software suite was developed using LabVIEW 6i with IMAQ-Vision 6.0.5 and the hardware employed included image capture cards (NI PCI-1408 and NI PCI-1409) and a DAQ card (NI PCI-6111E). The image acquisition module allows lossless recording of video data to a hard disk in real time, with analysis functions being available simultaneously. A subsequent part of the program allows more detailed analysis of the experimental data and prepares them for Þnal analysis in appropriate programs (MS Excel ® , Wavemetric Igor). Further modules of our software allow networking with other data sources (e.g., electrophysiological parameters, temperature, etc.) and the integration of control logic (e.g., controlling microscope motors). Other modules include procedures for image analysis in static ßuorescence microscopy with an auto-ROI function and an alignment procedure based on structure recognition for antidrift correction in prolonged sequences of images. 5.2.1 PROGRAM DESCRIPTION The modules described were integrated in the program on the basis of a menu-driven queued state machine and thus could be executed directly via the GUI, or as individual menu items, whereby a uniform variable space deÞnition ensures data exchange between the individual program components. 5.2.2 RECORDING VIDEO SEQUENCES For high spatial and temporal resolution recording of the dynamic changes in the ßuorescence characteristics of indicator dyes, a recording module (FastRecord) was developed to achieve the following: • Lossless, real-time display of the biological object under investigation on the computer monitor. In this case the (analogue) image delivered by the microscope is processed directly by the computer program. • Placement of ROIs over interesting regions of the image, interleaved with lossless, real-time display of brightness-versus-time courses. • Direct and lossless storage of image data on the hard disk. • Microscope control via the serial interface (e.g., laser shutter control, control of the z-drive). • Communication with the electrophysiological module, for instance via trigger connections (circuits) and networking of and with measurement data from other sources. • Simplicity and standardization of program operation, particularly with respect to appropriateness for the task in hand, capacity for self-description and conformity with expectation (in accordance with EN ISO 9241). • Modular software concept with the option of expansion of the program’s functionality at any time.
100 Image Acquisition Processing with LabVIEW The procedure in the course of an experiment begins with the biological object of interest (a single cell in a vital brain section) is displayed as a video image on the PC monitor. ROIs of any shape can be located at speciÞc positions in the image as desired. The mean brightness value of these ROIs can be displayed simultaneously beside the live image as brightness-versus-time courses on the monitor. If required, pressing a key stores image sequences, theoretically of any desired length, without loss and in real time. The only limit to the recording duration is the storage capacity of the hard disk. The possibility for direct-to-disk storage is a major advance, since this circumvents the limitations inherent in the transient and nonpermanent storage in the expensive RAM and thus allows direct and permanent storage of digital Þlms using the economic and theoretically unlimited capacity of the hard disk. In addition, it is possible to communicate with the various peripheral experimental instruments (laser shutter, microscope z-drive, etc.) and trigger connections allow data exchange with a so-called “patch-clamp” ampliÞer, so that the start of an electrophysiological experiment can initiate the recording of an image sequence and vice versa. 5.2.2.1 Analysis of Video Sequences The primary aim of the FastRecord module is the comfortable and comprehensive analysis of image sequences of various formats (AVI, Bio-Rad-PIC, BIN) obtained using FastRecord or other recording systems. The main focus thereby is the FIGURE 5.21 Calculate brightness vs. time. capacity to produce (after deÞnition of the appropriate ROIs) brightness-versus-time courses and to store the data in a format that can be read by conventional spreadsheet programs (Figure 5.21). The procedure for the calculation of the brightness-versus-time courses in calcium-imaging experiments is as follows: the mean brightness value (F) of a ROI is
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IMAGE PROCESSING SERIES Series Edit
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Foreword The introduction of LabVIE
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FIGURE 1 Vision 6.1 FIGURE 2 Compan
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Acknowledgments My gratitude goes o
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Dedication To Don “old fella” P
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Contents Chapter 1 Image Types and
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Chapter 4 Displaying Images........
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Chapter 7 Image Analysis...........
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1 Image Types and File Management F
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Image Types and File Management 3 F
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Image Types and File Management 5 A
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Image Types and File Management 7 C
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Image Types and File Management 9 p
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Image Types and File Management 11
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Image Types and File Management 13
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2 Setting Up You should never under
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Setting Up 17 FIGURE 2.3 (a) Interl
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Setting Up 19 Linear Aray Light Sou
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Setting Up 21 FIGURE 2.7 National I
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Setting Up 23 2.1.3.2 Control and I
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Setting Up 25 FIGURE 2.11 X-Ray ima
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Setting Up 27 Apple Computer techno
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Setting Up 29 FIGURE 2.17 An inexpe
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Setting Up 31 Although this would b
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Setting Up 33 134 -115 \ Contrast =
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Setting Up 35 resolution printer wi
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Setting Up 37 C A B C Traditional T
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Setting Up 39 Camera Light Source O
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Setting Up 41 Camera Light Source (
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Setting Up 43 FIGURE 2.37 (a) An ob
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Setting Up 45 FIGURE 2.41 (a) PCB p
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Morphology 149 FIGURE 6.8 Particle
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Morphology 151 FIGURE 6.12 Fill hol
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Morphology 153 FIGURE 6.17 Danielss
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Morphology 155 Detected circles wit
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Morphology 157 6.5 CASE STUDY: FIND
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Morphology 159 6.6 USER SOLUTION: D
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Morphology 161 ULTRASOUND MACHINE (
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Morphology 163 FIGURE 6.31 The Edit
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166 Image Acquisition Processing wi
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8 Machine Vision A generally accept
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Machine Vision 219 TABLE 8.1 “IMA
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Machine Vision 221 TABLE 8.3 Input
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Machine Vision 223 packages, includ
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Machine Vision 225 good sense to ad
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Machine Vision 227 TABLE 8.4 “IMA
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Glossary 1-Bit Image An image compr
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Glossary 231 binary display device
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Glossary 233 spatial Þlters, each
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Glossary 235 Resolution There are t
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Image Acquisitionand Proces

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