Supporting Information. Gold Nanoshell-Functionalized Polymer Nanoswimmer for Photomechanical Poration of Single Cell Membrane
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1 Supporting Information Gold Nanoshell-Functionalized Polymer Nanoswimmer for Photomechanical Poration of Single Cell Membrane Wei Wang, Zhiguang Wu*, Xiankun Lin, Tieyan Si, Qiang He* Key Laboratory of Micro-systems and Micro-structures Manufacturing (Ministry of Education), Harbin Institute of Technology, Yikuangjie 2, Harbin (China) * Video List Video S1: The movement of the AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening in pure water under acoustic field. (Frequency, 410 khz. Signal amplitude, 10 V p-p. Signal-amplifier, 10 db. Scale bar, 10 μm.) Video S2: Controlled forward/backward movement of the AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening under acoustic field. The time of regulating acoustic frequency is not included in the Figure 2C. (Frequency, 512/525 khz. Signal amplitude, 10 V p-p. Signal-amplifier, 10 db. Scale bar, 10 μm) Video S3: AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening rotating on the surface of the cell steadily for 106 s in acoustic field. Scale bar, 10 μm. Video S4: AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening moving towards to HeLa cell in acoustic field. Scale bar, 10 μm. Video S5: NIR irradiation (1.5 mj/μm 2 ) onto the AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening attached into the HeLa cell. Scale bar, 10 μm. Video S6: Dynamic intracellular fluorescence as the function of time after the NIR irradiation of the AuNSfunctionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening. The video is 10x accelerated. Scale bar, 10 μm. Video S7: AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening rotating on the surface of the cell for 2 s. The video is a fragment in Video S3 with 4x deceleration. Scale bar, 10 μm. S1
2 Figure S1. Setup of the optical microscope equipped with a home-made NIR laser. Figure S2. NIR irradiation treatment upon a gold-sputtering layer at a power of 1.5 mj/μm 2. The white ellipsoidal region represents the spot of NIR laser beam. Scale bar, 10 μm. S2
3 Figure S3. The UV-vis-NIR spectrum of the AuNS-functionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening. Figure S4. Statistics of the forward/backward movement of the AuNS-functionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening upon 10 cycles. F f, Forward frequency. F b, backward frequency. 1 cycle means a frequency change of (F f to F b ) and (F b to F f ). (A) Counts response of 10 cycles of frequency change (486 khz and 490 khz). (B) Velocity response of 10 cycles of frequency change (486 khz and 490 khz). S3
4 Figure S5. The forward/backward movement of two types of Au nanorod swimmers with diameters of 200 nm and 800 nm propelled with frequency change of acoustic field (486 khz and 490 khz). Scale bars, 10 μm. Figure S6. Comparative analysis of the effect of the wall thickness and assembling pairs of the AuNSfunctionalized polymer multilayer tubular nanoswimmers on their orientation propelled by an acoustic field. S4
5 Figure S7. Time-lapse images illustrating the end-on attachment of two small opening-orientated AuNSfunctionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening onto HeLa cells. Scale bar, 10 μm. Figure S8. Time-lapse images illustrating the end-on attachment of two small opening-orientated AuNSfunctionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening onto the same HeLa cell. Scale bar, 10 μm. S5
6 Figure S9. Enlarged view of the single AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening drilling on the surface of the cell membrane. Yellow dash line is the extending line of the vane on the big opening and red dash line indicates the rotation plane of the nanoswimmer's big opening. The angle illustrated in each image is obtained by measuring with the yellow dash line at 0 s. Scale bar, 10 μm. Figure S10. Schematic illustration for the critical force of the cell membrane poration. (A) AuNS-functionalized nanoswimmer could not open the cell membrane with ultrasound driving lonely because the driving force of ultrasound is much smaller than the critical force of cell membrane. (B) NIR laser was employed to assist the nanoswimmer in opening the cell membrane. S6
7 S7
8 Figure S11. Force analysis illustrating the angle change in the poration process. (A) The nanoswimmer deviated away from the perpendicular line of cell membrane surface after attaching the cell membrane. (B) A component (F a ) of photomechanical force (F propulsion ) vertically to the long axis from deviation could lead to an angle change in poration process. S8
9 Figure S12. The effect of different acoustic and NIR conditions on cell poration by the AuNS-functionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening. (A) Illumination area at the small opening of the nanoswimmer at a power of 1.5 mj/μm 2. (B) Illumination area at middle of the nanoswimmer at a power of 1.5 mj/μm 2. (C) Illumination for 25 s (15 mw/μm 2 ) without nanoswimmer. (D)Without acoustic field. laser power, 2 mj/μm 2. (E) Comparative analysis of the effect of each acoustic and NIR conditions. Red ellipsoidal region illustrates the region of NIR laser. Scale bars, 10 μm. S9
10 Figure S13. Shape and size effect on cell poration at a laser power of 1.5 mj/μm 2. (A) Au nanorod with a diameter of 200 nm. (B) Au nanorod with a diameter of 800 nm. (C) AuNS-functionalized (PSS/PAH) 10 swimmer with a diameter of 1 μm. (D) Comparative analysis of the effect of shape and size. Scale bars, 10 μm. S10
11 Figure S14. NIR irradiation treatment upon the big opening of the AuNS-functionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening on the cell membrane with different laser power. (A) 1 mj/μm 2. (B) 1.25 mj/μm 2. (C) 1.5 mj/μm 2. (D) Comparative analysis of the effect of different laser power. Scale bars, 10 μm. S11
12 Figure S15. NIR irradiation (1.5 mj/μm 2 ) treatment upon the big opening of the AuNS-functionalized (PSS/PAH) 6 (A), (PSS/PAH) 10 (B), (PSS/PAH) 14 (C), and (PSS/PAH) 18 (D) nanoswimmers with a diameter of 200 nm at the small opening on the cell membranes. (E) Comparative analysis of the effect of different wall thickness. Scale bars, 10 μm. S12
13 Figure S16. NIR irradiation (1.5 mj/μm 2 ) treatment upon the big opening of the AuNS-functionalized (PAA/PDDA) 10 (A) and (ALG/CHI) 10 (B) nanoswimmers with a diameter of 200 nm at the small opening on the cell membranes. (C) Comparative analysis of the effect of different composition. Scale bars, 10 μm. S13
14 Figure S17. NIR irradiation (1.5 mj/μm 2 ) treatment upon the two AuNS-functionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening on different cells successively. Scale bars, 10 μm. Figure S18. NIR irradiation (1.5 mj/μm 2 ) treatment upon the two AuNS-functionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening on the same cell successively. Scale bars, 10 μm. S14
15 Figure S19. The maximal splashing diameter of the ejecting superheat water by AuNS-functionalized (PSS/PAH) 10 nanoswimmers with a diameter of 200 nm at small opening upon different power of NIR irradiation at water/air interface. Scale bars, 10 μm. Figure S20. Dependence of the maximal splashing diameter on the energy of NIR irradiation. The energy of NIR irradiation is calculated by product of laser power density, irradiated area and irradiation time. S15
16 Figure S21. Computational simulation illustrating the temperature distribution around the AuNS-functionalized (PSS/PAH) 10 nanoswimmer with a diameter of 200 nm at small opening. Figure S22. Schematic illustrating the comparison of the NIR-assisted photothermal force and the critical force of cell poration. The inserted image illustrates the photothermal force distribution around the AuNSfunctionalized nanoswimmer upon the exposure of NIR light. The blue vectors indicate the force generated by temperature gradient. The size of the blue vector is proportional to the magnitude of force. The direction of the blue vector labels the orientation of local force vectors. F p is the resultant force and theoretically sufficient to open the cell membrane. S16
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