3D membrane segmentation and quantification of intact thick cells using cryo soft X-ray transmission microscopy: A pilot study

• DOI: 10.1371/journal.pone.0174324
• Publication Date: 2017-04-04T21:06:30Z
• Authors: Rubén Cárdenes; Chong Zhang; Oxana Klementieva; Stephan Werner; Peter Guttmann; Christoph Pratsch; Josep Cladera; Bart H. Bijnens
• Keywords: Cryopreservation; 0301 basic medicine; Microscopy; Phantoms, Imaging; Science; X-Rays; Q; Cell Membrane; R; Citologia; Large scale facilities for research with photons neutrons and ions; Pilot Projects; Cell Line; Cell membranes; Radiography; 03 medical and health sciences; Imaging, Three-Dimensional; Radiografia; Medicine; Humans; Membranes cel·lulars; Algorithms; Software; Research Article

Structural analysis of biological membranes is important for understanding cell and sub-cellular organelle function as well their interaction with the surrounding environment. Imaging whole cells in three dimension at high spatial resolution remains a significant challenge, particularly thick cells. Cryo-transmission soft X-ray microscopy (cryo-TXM) has recently gained popularity to image, 3D, intact (∼10μm) details architecture organization near-native state. This paper reports new tool segment quantify structural changes 3D from cryo-TXM images by tracking an initial 2D contour along third axis microscope, through multi-scale ridge detection followed active contours-based model, subsequent refinement other two axes. A quantitative metric that assesses grayscale profiles perpendicular membrane surfaces introduced shown be linearly related thickness. Our methodology been validated on synthetic phantoms using realistic microscope properties structure dimensions, real data. Results demonstrate validity our algorithms data analysis.