Tomographic phase microscopy of living three-dimensional cell cultures

• Published in: Journal of biomedical optics Vol. 19; no. 4; p. 046009
• Main Authors: Kuś, Arkadiusz, Dudek, Michał, Kemper, Björn, Kujawińska, Małgorzata, Vollmer, Angelika
• Format: Journal Article
• Language: English
• Published: United States Society of Photo-Optical Instrumentation Engineers 01.04.2014
• Subjects: Biotechnology; Cell Culture Techniques - methods; Cell Line, Tumor; Clusters; Digital; Equipment Design; Holography - methods; Humans; Microscopy; Optical fibers; Organelles; Refractometry; Three dimensional; Tomography; Tomography, Optical - methods; optical diffraction tomography; cancer cells; live cell imaging; digital holographic microscopy; refractive index tomography
• ISSN: 1083-3668; 1560-2281
• DOI: 10.1117/1.JBO.19.4.046009

A successful application of self-interference digital holographic microscopy in combination with a sample-rotation-based tomography module for three-dimensional (3-D) label-free quantitative live cell imaging with subcellular resolution is demonstrated. By means of implementation of a hollow optical fiber as the sample cuvette, the observation of living cells in different 3-D matrices is enabled. The fiber delivers a stable and accurate rotation of a cell or cell cluster, providing quantitative phase data for tomographic reconstruction of the 3-D refractive index distribution with an isotropic spatial resolution. We demonstrate that it is possible to clearly distinguish and quantitatively analyze several cells grouped in a "3-D cluster" as well as subcellular organelles like the nucleoli and local internal refractive index changes.