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Microscopy101


Snowfl akes . Microscopic documentation of snow crystals with their multitude of shapes has developed into a particular eye-catcher ( Figure 5 ), although preparation of such objects involves numerous complications and therefore requires a high degree of experience [ 7 , 8 ]. Snow crystals are distinguished from the other items described here by their platelet-like geometries, where one dimension ( z ) is signifi cantly reduced with respect to the other two ( x , y ). T is circumstance requires the production of raw images according to the stacking method. T ree-dimensional imaging of the crystals measuring several hundred micrometers in size may be used for a more compre- hensive exploration of the internal structure or the comparison of thicknesses developed in variably shaped objects.


Conclusion


Figure 5 : Stereophotograph of snow crystals developing different hexagonal geometries. Because of the fl atness of these objects, production of raw images with the stacking method illustrated in Figures 1 and 2 is highly recommended. Microscope: ZEISS Primotech, 10×, NA ~ 0.25. Image width = 2 mm. Semi-images are copyright of and used with permission of Eberhard Raap, micropolitan.org.


viewable with modest magnifi cations. T e preparation of rocks containing such fossils (radiolarites), however, turns out to be rather time-consuming because the shells have to be corroded out of the stony material with the help of hydrogen peroxide. Neurons .


We describe here two simple methods for the production of stereo images in transmitted light microscopy. Whilst the fi rst technique, conventional stereo pairs, is chiefl y restricted to isometric items of intermediate size (100 µm to 5 mm), the second technique, image stacks, is appropriate for small objects of any shape and size down to a few micrometers in extent. From the 3D micrographs exhibited here, it may be concluded that stereoscopic imaging represents a microscopy method whose scientifi c potential is not exhausted. In some research disciplines, the method is well-established, while in others it has not yet been explored.


Another application for stereo photomi-


crography is in biological tissue studies. Figure 4 shows the 3D view of the brain of a rat, underlining the signifi cance of this technique for microanatomical investigations. Here the production of stacked images with individual focus planes is highly recommended because of the limited thickness (30 µm) of the histological section. For an optimization of the spatial resolution, distances between images in the stack should not exceed 2 µm. Single dendritic cells such as the one in the center of the anaglyphic photograph have a diameter of about 20 µm and therefore have to be documented at higher magnifi cations. With the help of 3D images, courses of single dendrites and axons can be traced more easily, and the 3D arrangement of single nerve cells within the brain can be measured.


References [1] C Wheatstone , Transactions of the Royal Society of London 128 ( 1838 ) 371 – 94 .


[2] D Brewster , T e stereoscope: Its history, theory, and construction . J. Murray , London , 1856 .


[3] R Sturm , Mikrokosmos 97 ( 2 ) (2008 ) 75 – 80 . [4] R Sturm , Deposits Magazine 18 ( 2009 ) 10 – 13 . [5] R Sturm , Mikrokosmos 98 ( 6 ) (2009 ) 331 – 36 . [6] R Sturm , Deposits Magaz ine 26 ( 2011 ) 12 – 15 . [7] R Sturm , Mikroskopie 3 ( 2 ) (2016 ) 86 – 100 . [8] E Raap and H Cypionka , Mikrokosmos 100 ( 3 ) (2011 ) 140 – 44 . [9] R Sturm , Biologie in unserer Zeit 45 ( 1 ) (2015 ) 52 – 55 . [10] R Sturm , Naturwissenschaſt liche Rundschau 70 ( 4 ) (2017 ) 500 – 06 .


[11] JI Goldstein et al ., Scanning Electron Microscopy and X-ray Microanalysis . Springer , New York , 2003 .


2017 July • www.microscopy-today.com


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