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DDC: 610 × Subject: Fluorescence microscopy ×

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    Improved imaging of magnetically labeled cells using rotational magnetomotive optical coherence tomography
    (Basel : MDPI AG, 2017) Cimalla, P.; Walther, J.; Mueller, C.; Almedawar, S.; Rellinghaus, B.; Wittig, D.; Ader, M.; Karl, M.O.; Funk, R.H.W.; Brand, M.; Koch, E.
    In this paper, we present a reliable and robust method for magnetomotive optical coherence tomography (MM-OCT) imaging of single cells labeled with iron oxide particles. This method employs modulated longitudinal and transverse magnetic fields to evoke alignment and rotation of anisotropic magnetic structures in the sample volume. Experimental evidence suggests that magnetic particles assemble themselves in elongated chains when exposed to a permanent magnetic field. Magnetomotion in the intracellular space was detected and visualized by means of 3D OCT as well as laser speckle reflectometry as a 2D reference imaging method. Our experiments on mesenchymal stem cells embedded in agar scaffolds show that the magnetomotive signal in rotational MM-OCT is significantly increased by a factor of ˜3 compared to previous pulsed MM-OCT, although the solenoid's power consumption was 16 times lower. Finally, we use our novel method to image ARPE-19 cells, a human retinal pigment epithelium cell line. Our results permit magnetomotive imaging with higher sensitivity and the use of low power magnetic fields or larger working distances for future three-dimensional cell tracking in target tissues and organs.
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    Optical Sectioning and High Resolution in Single-Slice Structured Illumination Microscopy by Thick Slice Blind-SIM Reconstruction
    (San Francisco, California, US : PLOS, 2015) Jost, Aurélie; Tolstik, Elen; Feldmann, Polina; Wicker, Kai; Sentenac, Anne; Heintzmann, Rainer; Degtyar, Vadim E.
    The microscope image of a thick fluorescent sample taken at a given focal plane is plagued by out-of-focus fluorescence and diffraction limited resolution. In this work, we show that a single slice of Structured Illumination Microscopy (two or three beam SIM) data can be processed to provide an image exhibiting tight sectioning and high transverse resolution. Our reconstruction algorithm is adapted from the blind-SIM technique which requires very little knowledge of the illumination patterns. It is thus able to deal with illumination distortions induced by the sample or illumination optics. We named this new algorithm thick slice blind-SIM because it models a three-dimensional sample even though only a single two-dimensional plane of focus was measured.