PubMed · 5940017
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L T Fitzgerald, F P Bruno, W Mauderli. 1966. An automatic depth-dose acquisition system.. https://doi.org/10.1148/86.6.1107
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Gel analysis system purchasers have a large variety of digital imaging products to choose from today. Understanding the specifications presented by the manufacturers is important to ensure that the system fits the needs ofthe researcher using the system. The background information contained in this paper explains the terminology and many of the specifications along with their significance.
Image capture is the first step of image analysis. There are two major devices for image capture in the field of electrophoresis. One is the charged-couple device (CCD) camera and the other is the scanner. Image capture technologies have shown great progress in recent years especially in the field of fluorescence detection and chemiluminescent detection. The direction of image analysis is high resolution, wide dynamic range and high density precision and this holds true for the CCD camera system. Various components in the CCD camera system suitable for high-sensitive fluorescence detection and chemiluminescent detection are explained. As an example, the LAS-1000plus camera system which has 1364 x 922 pixels and generates 14-bits image is introduced. Powerful cooling enables overnight exposure of chemiluminescence. Introduction of blue light-emitting diode (LED) as excitation light source improved safety to eyes. Two types of scanners for fluorescence detection and the specific characteristics are explained. There are mechanical scanning systems using confocal optics and optical scanning systems using light collecting guide optics. Deep focusing range and equal fluorescence intensity at various depth is a characteristic feature of light collecting guide optics.
In recent years the use of fluorescent dyes in biological applications has dramatically increased. The continual improvement in the capabilities of these fluorescent dyes demands increasingly sensitive detection systems that provide accurate quantitation over a wide linear dynamic range. In the field of proteomics, the detection, quantitation and identification of very low abundance proteins are of extreme importance in understanding cellular processes. Therefore, the instrumentation used to acquire an image of such samples, for spot picking and identification by mass spectrometry, must be sensitive enough to be able, not only, to maximise the sensitivity and dynamic range of the staining dyes but, as importantly, adapt to the ever changing portfolio of fluorescent dyes as they become available. Just as the available fluorescent probes are improving and evolving so are the users application requirements. Therefore, the instrumentation chosen must be flexible to address and adapt to those changing needs. As a result, a highly competitive market for the supply and production of such dyes and the instrumentation for their detection and quantitation have emerged. The instrumentation currently available is based on either laser/photomultiplier tube (PMT) scanning or lamp/charge-coupled device (CCD) based mechanisms. This review briefly discusses the advantages and disadvantages of both System types for fluorescence imaging, gives a technical overview of CCD technology and describes in detail a unique xenon/are lamp CCD based instrument, from PerkinElmer Life Sciences. The Wallac-1442 ARTHUR is unique in its ability to scan both large areas at high resolution and give accurate selectable excitation over the whole of the UV/visible range. It operates by filtering both the excitation and emission wavelengths, providing optimal and accurate measurement and quantitation of virtually any available dye and allows excellent spectral resolution between different fluorophores. This flexibility and excitation accuracy is key to multicolour applications and future adaptation of the instrument to address the application requirements and newly emerging dyes.