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This is because at high resolution, those objects cannot hold within the depth of field of a microscope and therefore a single 2D image acquisition often suffers from being only partially in focus. Image stacks are also useful to image flattish or so-called 2.5D objects such as an epithelium, a monolayered cell culture, a membrane within an in vivo tissue sample or a flat biological structure such as cultured neurons. A variety of software programme as Voxx 6, NIH’s Fiji/ImageJ 7 or VTK 8 propose such options with an interactive visualization. ![]() Optionally, a detection step can be introduced before the projection to focus on the reconstruction of layers of interest 5. In this case, a single projection cannot be satisfactory and an interactive rendering tool must be used to obtain successive projections of the data in arbitrary directions using transparency filters. Unlike a single 2D image, which can be rendered and interpreted straightforwardly on a screen, a 3D stack needs to be processed to be displayed.Īn image stack can be employed to image a full 3D object. Furthermore, it is common nowadays to image the same preparation repetitively at various depths to obtain a three-dimensional (3D) image under the form of a stack of two-dimensional (2D) images. The image can then be saved as a file or shared with others after you’ve cropped it in the way you want.Cell biologists routinely use fluorescence microscopy to observe spatial organization of organelles, co-localization of vesicles or to study the impact of gene expression or small compounds on cell morphology 1, 2, 3, 4. In the resize and rotate options, you can change the angle of the crop area as well as size and size of the crop area. If you want to use the desired effect, select the crop area and then select the toolbar options. 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