The maker movement has reached the optics labs, empowering researchers to actively create and modify microscope designs and imaging accessories. 3D printing has especially had a disruptive impact on the field, as it entails an accessible new approach in fabrication technologies, namely additive manufacturing, making prototyping in the lab available at low cost. Examples of this trend are taking advantage of the easy availability of 3D printing technology. For example, inexpensive microscopes for education have been designed, such as the FlyPi. Also, the highly complex robotic microscope OpenFlexure represents a clear desire for the democratisation of this technology. 3D printing facilitates new and powerful approaches to science and promotes collaboration between researchers, as 3D designs are easily shared. This holds the unique possibility to extend the open-access concept from knowledge to technology, allowing researchers from everywhere to use and extend model structures. Here we present a review of additive manufacturing applications in microscopy, guiding the user through this new and exciting technology and providing a starting point to anyone willing to employ this versatile and powerful new tool.
Fluorescence microspectroscopy (FMS) with environment-sensitive probes provides information about local molecular surroundings at microscopic spatial resolution. Until recently, only probes exhibiting large spectral shifts due to local changes have been used. Herein, we show that appropriate measuring procedure and data analysis enable nanometer spectral peak position resolution, even for photosensitive fluorophores [1]. The reach of our approach is demonstrated in several examples. The first application shows how we can distinguish lipid vesicles in different lipid phases with two commonly used polarity-sensitive probes. A synthesized NBD-based fatty acid red-shifted its emission maximum by 1.5-2 nm going from gel to liquiddisordered phase in DPPC. Between these two phases Laurdan exhibits a large 50 nm red-shift. We therefore chose a more challenging combination -gel and liquid ordered phase, realized by DPPC and DPPC/Chol (40 mol%), respectively, where we were able to detect a 3 nm blue-shift with Laurdan [1]. The second example shows application of a synthesized rhodamine-based pH-activatable probe that is sensitive to aggregation. We studied a receptor-mediated internalization in dendritic cells and measured a 3 nm aggregation-induced emission spectral shift due to probe accumulation in endosomes and lysosomes [2,3]. The results show that peak position resolution, characteristic for spectrofluorimetric measurements on bulk samples, could readily be achieved at micrometer spatial scale.[1] I. Urban ci c, Z.
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