The way to determine if a picture of a specimen is according to the right parameters and can be considered to be a ‘good’ and ‘useful’ picture is when the following settings have been met: i) the image needs to have an in-focus specimen ii) there shouldn’t be any parts that are over-exposed or under-exposed iii) all parts that are necessary to identify/study a specimen in a specific view have to be visible and distinguishable.
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These systems could be relatively fast, but often, post-processing was time consuming and most importantly, as techniques change rapidly, an update on the hardware of these systems is quite expensive. Although results were better than a single image, the system itself was sometimes difficult to operate without a training period. In the beginning this technique was only available for laboratories or research groups with a large budget to spend on a state of the art stacking column or microscope, with special lenses, lighting, stage control, camera and software. 1984, CombineZP ( Helicon Focus ( Auto-Montage ( Zerene Stacker ( /cms/home)).
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As computers have become widely available, numerous software have been developed by microscope companies and researchers, making it possible to combine pictures with different depths of field to create a focus stack in which the entire object is in focus ( Adelson et al. However this resulted in other aberrations as the optical resolution reduces due to the diffraction effect ( Davies 2010, Gallo et al. The low depth of field made it almost impossible to get the complete object in focus, unless the aperture was stepped down ( Tindall and Kalms 2012). When suitable lenses made it possible to capture small creatures on film it was clear that other problems arose. This has always worked for regularly sized objects however, the micro-world remained unrecorded for a long time. Since the first photographic equipment was developed, people have tried to record natural history specimens with their equipment. Overall, this enables institutions to purchase multiple solutions or to start digitising the type collection on a large scale even with a small budget. The largest benefit is the price of the set-up which is approximately € 3,000, which is 8 and 10 times less than the LeicaZ6APO and LeicaMZ16A set-up respectively.
The flash lighting inside the Ikea closet creates an evenly illuminated picture, without struggling with filters, diffusers, etc. The resolution of the tested extended focus pictures is much higher than those from the Leica systems. The outcome of the test on high-end solutions demonstrates that our approach performs better in several ways. However, Zerene Stacker gives the user more possibilities in terms of control of the software, batch processing and retouching.
Zerene Stacker and Helicon Focus both provided satisfactory results. We tested our final stacked picture with a picture obtained from two high-end focus stacking solutions: a Leica MZ16A with DFC500 and a Leica Z6APO with DFC290. We tested this system and compared the results with several different software packages (CombineZP, Auto-Montage, Helicon Focus and Zerene Stacker). The system is inexpensive compared to high-end commercial focus stacking solutions. In this manuscript we present a focus stacking system, composed of commercial photographic equipment.
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