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. 2018 Sep 21;8:14191. doi: 10.1038/s41598-018-32578-w

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© The Author(s) 2018

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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Experimental set up for multiple radiosensitizations in vivo. (A) Device i. In order to reduce toxicity during multiple radiosensitization cycles, CED was carried out under inhalation anesthesia with isoflurane as described under Materials and methods and¹³. Top left and top central insets are detail snapshots showing the location of CED guide screw on the mouse skull with distances from the left ear and eye and the simultaneous inhalation anesthesia of three mice, respectively. (B) Device ii: the RS 2000 Biological Irradiator (Rad Source Technologies, Alpharetta, GA, USA) equipped with the inhalation anesthesia device. (C) Device iii: the irradiation chamber with inhalation anesthesia tubing. (D) Device iv: the X-ray collimator with positioning laser beams. (E) Dose calibration i: the mouse polystyrene support with RadCal X-ray calibrating probe. (F) Dose calibration ii: the RadCal dosimeter display. (G) Dose calibration iii: radiochromic films placed under and over the mouse body and positioning the X-rays beam to the brain by the collimator laser. (H) Dose calibration iv: the upper radiochromic film before irradiation. (I) Dose calibration v- the upper radiochromic film after irradiation. The irradiated region of top head is indicated by the darkened region of the radiochromic film. (J) Dose calibration vi: example of dose distribution quantification after scanning of radiochromic film. (K) Dose calibration vii: the irradiated region under the head as indicated by the darkened region of the bottom radiochromic film. (L) Dose calibration viii: the 70°-rotated animal allowing direct irradiation of the left, tumor-bearing brain hemisphere. (M) Dose calibration ix: as in K but with high-sensitivity radiochromic film allowing visualization of the guide screw position (indicated with arrow). (N) Dose calibration x: magnification of L with site of guide screw indicated with arrow. (O) Dose calibration xi: example of RS 2000 parameters for irradiation with 0.5 Gy to the head and virtually 0.0 Gy to the body.