Confocal microscopy is widely used for three-dimensional (3D) sample reconstructions. Arguably, the most significant challenge in such reconstructions is posed by the resolution along the optical axis being significantly lower than in the lateral directions. In addition, the imaging rate is lower along the optical axis in most confocal architectures, prohibiting reliable 3D reconstruction of dynamic samples. Here, we demonstrate a very simple, cheap, and generic method of multiangle microscopy, allowing high-resolution high-rate confocal slice collection to be carried out with capillary-contained colloidal samples in a wide range of slice orientations. This method, realizable with any common confocal architecture and recently implemented with macroscopic specimens enclosed in rotatable cylindrical capillaries, allows 3D reconstructions of colloidal structures to be verified by direct experiments and provides a solid testing ground for complex reconstruction algorithms. In this paper, we focus on the implementation of this method for dense nonrotatable colloidal samples, contained in complex-shaped capillaries. Additionally, we discuss strategies to minimize potential pitfalls of this method, such as the artificial appearance of chain-like particle structures.
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The authors are grateful to Y. Rabin, D. Osmanovic,́ A. Nikolaenkova, M. Hermes, and W. Vlug for fruitful discussions, to M. Izoh, G. Gershinsky, and P. Helfferich for technical assistance, and to the Kahn foundation for the purchase of equipment. Acknowledgment is made by S.R.L., G.I., A.V.B., and E.S. to the donors of the American Chemical Society Petroleum Research Fund for support of this research. P.J.L. and D.A.W. thank NASA (NNX13AQ48G) for the financial support. A.v.B. and E.B.v.d.W. acknowledge the European Research Council (ERC) under the European Union’s Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement 291667.
© 2017 American Chemical Society.