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. 2017 Jun 23;7:4165. doi: 10.1038/s41598-017-04220-8

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The OCM system and experimental layout. (a) Imaging setup: the original parts of the inverted microscope are marked in blue and custom OCM parts in black. (b) Schematic of image cross-sections obtained with the OCM setup. (c) OCM beam trajectory in the XY plane. Solid line: the beam trajectory during data acquisition. Dotted line: the beam trajectory between Y positions. (d) Schematic of the data volume acquired in a single OCM measurement. The A-scan is a single image line obtained from one OCM data acquisition. The B-scan is a single image generated from the A-scans. The V-scan is generated from the B-scans acquired at different Y positions. (e–g) Stages of the diversified time interval scanning protocol (DTIsp). (e) Several V-scans are acquired within ~1 sec and undergo temporal averaging. (f) The averaged V-scans from (e) were acquired continuously with a time span ranging from ~20–600 sec for periods of tens of hours. (g) Temporal averaging (TA) was performed on the averaged V-scans from (f) to obtain images with reduced speckle noise. (h) Optional spatial averaging (SA) over thick slices of the averaged V-scan. Different types of averaging are possible, including intensity averaging (mean), minimal or maximal intensity projections (MIP). (i) Images from consecutive steps of DTIsp processing. Linear intensity mapping allows for a better visualization of the cytoplasmic features, while logarithmic - of the nuclei. (j) Cell segmentation and rendering.