Detection and Image Analysis
The signal from a blot can be captured using autoradiography films, storage phosphor screens, or image acquisition systems, depending upon the labeling method and the required levels of sensitivity and resolution.
Autoradiography Films are photographic films that can capture the images created by electromagnetic radiation, or particles emitted by radioisotopes. They can be used to detect radiation from radioactively-labeled probes and also the light generated by, for example, chemiluminescence methods. Results obtained with this method can be scanned and analyzed using image analysis software. Autoradiography films are inexpensive, reliable, and provide a good level of sensitivity and resolution, but need development chemicals and access to a darkroom.
Image Acquisition Systems such as CCD cameras and laser-based scanners are usually used with chemiluminescence, chemifluorescence, and fluorescence labeling methods. Some scanners are also able to acquire radioactive signals using a reusable storage phosphor screen, which captures latent images produced by ionizing radiation, such as emissions from radioisotopes. When stimulated with a laser, light is emitted from the storage phosphor screen in proportion to the amount of radioactivity in the sample and can be detected. Storage phosphor screens avoid the need for autoradiography film, development chemicals, and a darkroom. They are very sensitive and may require only one tenth of the exposure time required by autoradiography film.
Image Quant TL Analysis Software is a flexible, easy-to-use software package for the quantitative analysis of images from a wide range of biological samples. ImageQuant TL is designed for a broad range of applications including 1-D electrophoretic separations, dot blot, slot blot, microplate, TLC, macroarray, and plaque assays. Its high level of automation and throughout allows for automatic analysis of 1-D gels and blots including lane creation, background subtraction, band detection, molecular weight calibration, quantity calibration, and normalization. Analysis reports are dynamically linked to Microsoft Excel™.