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How to Do Everything: Digital Camera (5th ed.). ^ "Malin Space Science Systems - Mars Science Laboratory (MSL) Mars Hand Lens Imager (MAHLI) Instrument Description".
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Unix, Linux, Windows, Mac OS 9 and Mac OS X ImageJ with Extended Depth of Field PluginĪlex Prudencio, Jesse Berent, Daniel Sage Shutter Stream Product Photography Software Software / Application Focus stacking softwareĪmira / Avizo 'Image Stack Projection' Įnfuse (combined with align_image_stack or similar) By taking a through-focal series, the depth of focus can be reconstructed to create a single image entirely in focus. When observing a sample directly the limitations of the shallow depth of field are easy to circumvent by focusing up and down through the sample to effectively present microscopy data of a complex 3D structure in 2D, focus stacking is a very useful technique.Ītomic resolution scanning transmission electron microscopy encounters similar difficulties, where specimen features are much larger than the depth of field.
Higher magnification objective lenses generally have shallower depth of field a 100× objective lens with a numerical aperture of around 1.4 has a depth of field of approximately 1 μm.
A high numerical aperture (equivalent to a low f number) gives a very shallow depth of field. In microscopy high numerical apertures are desirable to capture as much light as possible from a small sample. The Mars Science Laboratory mission has a device called Mars Hand Lens Imager (MAHLI), which can take photos that can later be focus stacked. The hole is 1.6 centimetres (0.63 in) wide and 6.7 centimetres (2.6 in) deep. Stacked image of the Curiosity Rovers first sampling hole in Mount Sharp. Alternative techniques for generating images with increased or flexible depth of field include wavefront coding and light-field cameras.
Focus stacking also allows generation of images physically impossible with normal imaging equipment images with nonplanar focus regions can be generated. Focus stacking can also be useful in landscape photography.įocus stacking offers flexibility: since it is a computational technique, images with several different depths of field can be generated in post-processing and compared for best artistic merit or scientific clarity. Focus stacking can be used in any situation where individual images have a very shallow depth of field macro photography and optical microscopy are two typical examples. Bottom is the final "focus stacked" image.įocus stacking (also known as focal plane merging and z-stacking or focus blending) is a digital image processing technique which combines multiple images taken at different focus distances to give a resulting image with a greater depth of field (DOF) than any of the individual source images.
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Top right are the three masks used to obtain the contributions of their respective images to the final "focus stacked" image (black is no contribution, white is full contribution). Top left are the three source images captured at three different focus distances. This example is of a diatom microfossil in diatomaceous earth. Focus stacking (for extended depth of field) in bright field light microscopy.
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