Three-Dimensional Printing
of extruder head temperature. Ultimately we were unable to fi nd a temperature that sustained printing over the time required without clogging or stopping altogether. T is led us to the dissolvable fi lament, which was much more tractable. Interestingly, we also had varying degrees of success with diff erent colors of ABS fi lament, certain colors being much more prone to clog than others. Finally, the conversion of multiple fi le types proved troublesome. While none of the currently available image processing and acquisition soſt ware packages off er an .stl fi le format export option, we are optimistic that with the rise in popularity of 3D printing, this is not far off .
In accordance with the open spirit of many of the maker communities, there are a number of openly accessible repositories for 3D print fi les. T ese include the MakerBot T ingiverse, which features more than 400,000 designs ( thingi-
verse.com ). With specifi c scientifi c focus, the NIH 3D print exchange (
http://3dprint.nih.gov ) was launched in June 2014 in order to coincide with the fi rst White House Maker Faire. T e site off ers a repository for models and laboratory equipment, as well as instructional videos and a discussion forum. In order to share our design with others, the .stl fi les comprising our large actin model were deposited on the NIH 3D print exchange website and are available at
http://3dprint.nih.gov/ discover/3dpx-000748 .
Figure 4 : Completed F-actin models: the larger 20 cm model was printed in quarters (top), and the 10 cm models were printed in various colors (bottom). Scale bar (top)=5 cm on model; scale bar=5 μ m on original STED image.
base required a three-hour session in a 2L D-limonene bath, followed by cleaning with soap and water to reduce the fairly oily (yet pleasantly scented) residue leſt by limonene. T e fi nal result was a sturdy yet detailed model of the NK cell F-actin cytoskeleton ( Figure 4 ).
Discussion T e greatest troubleshooting aspect of this procedure was the generation of fi les that were not too complex, particularly given the nature of the cytoskeleton our model was based on. Despite this, we estimate that our printed model gives a resolution of 75 nm at its most detailed points, particularly the actin fi laments extending from the cell edge. Rendering at both the Imaris and Blender stages was prone to freezing and crashing our relatively high-powered PC computers if too much detail was retained in the image. In addition, although we had few problems with ABS fi lament material, switching to polylactic acid (PLA) polymer in an attempt to use opaque- colored fi lament as a base required signifi cant optimization
2015 July •
www.microscopy-today.com
Conclusion T e current boom in 3D printing technology is giving rise to a number of creative and innovative applications. T e ability to feel the structures that we see in a micrograph is more than a cute exercise. It provides us the ability to add a tactile component to our thinking about the cell structures and biological processes in which we are engaged. Further, it provides a touchstone through which we can share our research and engage the public. As researchers working in a clinical setting, this provides an important way to bring the seemingly intangible ideas of immune function to those who are not immunologists or cell biologists. T is could include a powerful experience for the blind or the visually impaired. T rough our combination of non-diff raction limited microscopy with 3D printing, we have made small cell structures not just visible, but also touchable.
Acknowledgments T e authors wish to thank Dr. T. Feinstein for the initial dialogue that led to this article. T is work is supported by the 3D Print Program of the Center for Human Immunobiology at Texas Children’s Hospital.
References [1] D Toomre and J Bewersdorf , Annu Rev Cell Dev Biol 26 ( 2010 ) 285 – 314 .
[2] EM Mace and JS Orange , Front Immunol 3 ( 2012 ) 421 . [3] K Lagrue et al ., Immunol Rev 256 ( 1 ) ( 2013 ) 203 – 21 . [4] JS Orange , J Allergy Clin Immunol 132 ( 3 ) ( 2013 ) 515 – 25 , quiz 526.
[5] GD Rak et al ., PLoS Biol 9 ( 9 ) ( 2011 ) e1001151 . [6] AC Brown et al ., PLoS Biol 9 ( 9 ) ( 2011 ) e1001152 .
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