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. 2017 Dec 19;7:17769. doi: 10.1038/s41598-017-18227-8

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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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Effects of laser power and PMT voltages on fluorescence and SSC. MLVeGFP was analysed with systematic adjustments of voltage for detection of green fluorescence and SSC off the 488 nm laser. (A) Increments of 50 V were made on SSC off the 488 nm laser from 225 V to 425 V while maintaining a consistent MFI for the eGFP+ population on the green fluorescence channel (488nm-530/30), while simultaneously increasing laser power from 50 to 300 mW. Threshold was set at SSC fluorescence intensity 200. (B) The same voltage and laser power adjustments were made on the green fluorescence channel while maintaining constant the MFI in SSC of the eGFP+ population. (C) Tabulation of the number of eGFP+ events in gates set in A. (D) Tabulation of the number of electronic aborts generated during the acquisition of each condition described in A. (E) Fluorescence Index calculated for each condition portrayed in B. FL-index = (MFI_eGFP+ − MFI_eGFP−)/SD_eGFP−. Data is representative of three separate experiments.