Special Section on Advanced Laser Technologies for Biophotonics

Axial tomography in live cell laser microscopy

[+] Author Affiliations
Verena Richter, Sarah Bruns, Thomas Bruns, Petra Weber, Michael Wagner, Herbert Schneckenburger

Aalen University, Institute of Applied Research, Beethovenstraße 1, 73430 Aalen, Germany

Christoph Cremer

University of Heidelberg, Institute of Pharmacy and Molecular Biology, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany

Institute of Molecular Biology, Ackermannweg 4, 55128 Mainz, Germany

J. Biomed. Opt. 22(9), 091505 (Jan 25, 2017). doi:10.1117/1.JBO.22.9.091505
History: Received October 11, 2016; Accepted January 3, 2017
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Abstract.  Single cell microscopy in a three-dimensional (3-D) environment is reported. Cells are grown in an agarose culture gel, located within microcapillaries and observed from different sides after adaptation of an innovative device for sample rotation. Thus, z-stacks can be recorded by confocal microscopy in different directions and used for illustration in 3-D. This gives additional information, since cells or organelles that appear superimposed in one direction, may be well resolved in another one. The method is tested and validated with single cells expressing a membrane or a mitochondrially associated green fluorescent protein, or cells accumulating fluorescent quantum dots. In addition, axial tomography supports measurements of cellular uptake and distribution of the anticancer drug doxorubicin in the nucleus (2 to 6 h after incubation) or the cytoplasm (24 h). This paper discusses that upon cell rotation an enhanced optical resolution in lateral direction compared to axial direction can be utilized to obtain an improved effective 3-D resolution, which represents an important step toward super-resolution microscopy of living cells.

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© 2017 Society of Photo-Optical Instrumentation Engineers

Citation

Verena Richter ; Sarah Bruns ; Thomas Bruns ; Petra Weber ; Michael Wagner, et al.
"Axial tomography in live cell laser microscopy", J. Biomed. Opt. 22(9), 091505 (Jan 25, 2017). ; http://dx.doi.org/10.1117/1.JBO.22.9.091505


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