- #IMAGEJ WESTERN BLOT QUANTIFICATION MANUAL#
- #IMAGEJ WESTERN BLOT QUANTIFICATION SOFTWARE#
- #IMAGEJ WESTERN BLOT QUANTIFICATION FREE#
The data gained from image analysis are exported by ImageJ as numerical values that can be further processed in software packages for statistical analysis. One function of ImageJ is the recording of macros: 1, 3 different procedures that can be applied to images, are recorded in a Java-like programming language, can be customized and later applied to a random number of other images. The software may not that easy to be used straightaway, but after a short time of training, it will become a versatile and adaptable tool that does not rank behind (still very expensive) commercial packages.
#IMAGEJ WESTERN BLOT QUANTIFICATION FREE#
For such (as far as possible) automatized large-scale image analysis, the free software “ImageJ” 2 has become a valuable and highly flexible tool for researchers.
#IMAGEJ WESTERN BLOT QUANTIFICATION MANUAL#
Thus, an automatized solution is necessary, that requires only minimal manual input, customization, and time for analysis of large image stacks. Manual evaluation of hundreds of images, containing thousands of cells per image, as found in many examples (here: tissue slices from mouse liver), is not practicable and may also result in subjective evaluation (human bias) by the researcher. To detect small shifts of cellular proteins, large numbers of cells from different samples have to be analyzed and compared in order to gain statistic validity. Acquisition of large amounts of highly resolving, multi-channel fluorescence- or confocal microscopic images is meanwhile also a quite automatized process, quickly producing huge stacks of images that may contain several thousands of individual cells, each. Abstractįluorescence/confocal microscopy is one of the main methods investigating intracellular distributions of immunostained proteins. “Cyt/Nuc” is easy to use and highly customizable, matches the precision of careful manual evaluation and bears the potential for quick detection of any shift in intracellular protein distribution. A significant shift in proteasomal distribution between cytosol and nucleus in response to metabolic stress was revealed using “Cyt/Nuc” via automatized quantification of thousands of nuclei within minutes. As practical example, the redistribution of the 20S proteasome, the main intracellular protease in mammalian cells, is investigated in NZO-mouse liver after feeding the animals different diets. In this work, we present “Cyt/Nuc,” an ImageJ macro, able to recognize and to compare the nuclear and cytosolic areas of tissue samples, in order to investigate distributions of immunostained proteins between both compartments, while it documents in detail the whole process of evaluation and pattern recognition. Since ImageJ offers recording of “macros,” even a complex multi-step process can be easily applied fully automated to large numbers of images, saving both time and reducing human subjective evaluation. The freeware “ImageJ” has become one of the standard tools for scientific image analysis. Large amounts of data from multi-channel, high resolution, fluorescence microscopic images require tools that provide easy, customizable, and reproducible high-throughput analysis.
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