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75 mm × 25 mm microscopy slides (631-0113, VWR, Germany) are acclimated for the appraisal of the apery adjustment and abate 22 mm × 22 mm microscope coverslips (470055, Brand, Germany) as substrates for the abasement of ultra-thin sections. Afore the experiments, the substrates are blood-soaked in charwoman band-aid (2% Micro-90, Sigma-Aldrich, Germany) for 24 h and rinsed thoroughly with de-ionised (DI) water, followed by 10 min accelerated charwoman in acetone and isopropyl booze respectively. Subsequently, the substrates are rinsed with DI water, oven-dried for 2 h at 120 °C and stored in closed containers. Immediately afore the silanisation processes, the substrates are advised in an oxygen claret (Diener Atto, Germany) for 10 min at 0.5 mbar to access the cardinal of accessible OH-surface groups for siloxane bonding.



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Figure 2 illustrates the brand alertness process. Laser-cut stainless animate breadth metal (size of 22 mm × 22 mm, array of 200 µm, PCB affectation manufacturer, Beta LAYOUT, Germany), with the appearance of the ambition pattern, are acclimated as masters for hot embossing. The breadth metal is placed on polymethylmethacrylat (PMMA) bedding (Plexiglas, Evonik Industries, Germany) with 2 mm array and hot-embossed with an activated force of 2.2 kN in a automatic columnist at 125 °C for 1 h. The PMMA mould is again cooled bottomward to 80 °C aural an hour and annealed at that temperature for addition hour to abate stress. To facilitate afterwards demoulding of the final stamp, the consistent PMMA mould is advised with perfluorodecyltrichlorosilane (FDTS) (97%, Alfa Aesar, Germany). Poly(dimethylsiloxane)-based (PDMS) stamps (Sylgard 184® elastomer, Dow, Germany) are able by bond elastomer and abating abettor in a 10:1 weight arrangement and convalescent at 75 °C for 2 h in the PMMA moulds. Afterwards demoulding the PDMS is rinsed with heptane to abolish balance airy apparatus and broiled in air.

Laser-cut breadth metal acclimated for PMMA imprinting to actualize PMMA moulds to casting PDMS stamps.

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Vapour appearance silanisation: A 5% band-aid of 1,7-dichloro-octamethyltetrasiloxane (Cl[Si(CH3)2O]3Si(CH3)2Cl) in heptane (Sigmacote®, Sigma-Aldrich, Germany) is acclimated for the alertness of berserk coating. Complete silanisation of the substrate apparent is performed by alignment 10 microscope slides angular with 10 mm agreement in a closed alembic with a aggregate of 900 cm3. The alembic is placed in an oven preheated to 55 °C to acquiesce constant calefaction administration and again 150 µl of siloxane (5%) is added to accomplish breath appearance silanisation for 60 min at 55 °C. Subsequently, the substrates are rinsed with DI water.



Oxygen claret patterning: The silanised apparent is again covered with the PDMS stamps and apparent to an 13.56 MHz RF oxygen claret at 0.5 mbar (Diener Atto, Germany) for 15 minutes. The brand protects the siloxane band at the acquaintance surfaces and allows claret assay of the band in the 200 µm aerial cavities as illustrated in Fig. 3.

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Process accomplish of oxygen claret assay in cavities (I) and microcontact press (II) to actualize a blooming siloxane modification as a abasement template.

Microcontact printing: 1,7-dichloro-octamethyltetrasiloxane (95%, Sigma-Aldrich, Germany) is acclimated for microcontact press experiments. The PDMS stamps with an alien ambit of 22 mm × 22 mm are active with a afresh able band-aid of 10% chlorosiloxane in acetone (p. a., Merck, Germany) as illustrated in Fig. 3. The stamps are air-dried for 20 s and again brought into acquaintance with the bottle apparent for 60 s. Subsequently, the substrate is rinsed with DI water.

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The hydrophobicity of siloxane coatings created by vapour appearance silanisation is characterized with a bead appearance analyzer (DSA 100, KRÜSS, Germany) by barometer the beggarly changeless acquaintance bend (CA) amid distilled baptize and the substrate surface. Two abstracts application the sessile bead adjustment are performed on anniversary substrate surface. For the measurement, a atom aggregate of 10 µl is dispensed and the consistent curve fit to a Young-Laplace archetypal for acquaintance angles >10° and to a amphitheater archetypal for abate acquaintance angles. The baseline is set manually and abstracts are done 10 s, 30 s, 1 min and 2 min afterwards bead deposition.

In adjustment to appraise the aqueous ascendancy backdrop of the consistent approach templates, the hydrophilic channels are abounding with an accretion bulk of baptize and inspected optically. The aqueous basin is placed abutting to the alpha of the channels and boring (10 µl/min) abounding with DI baptize application a syringe pump (NE-1000, New Era Pump Systems Inc, USA). Images are taken at 0.88 µm resolution with an optical microscope (INM 200, Leica, Germany). Application the refraction at the baptize apparent in top illumination, overextension of the wetting aqueous into the wettable approach breadth is acutely visible, acceptance automatic assay with angel processing accoutrement (canny bend apprehension algorithm, scikit-image library). The curve of the advance aqueous is fit adjoin a beeline nominal appearance to appraisal the breadth and amplitude of the hydrophilic channels. The basis of the boxlike applicable absurdity (standard deviation) is acclimated to appraisal the aberration from the nominal shape. The channels are again abounding with baptize until the applicable absurdity increases steeply, which indicates the best baptize accumulator capacity.

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Time-of-Flight Accessory Ion Accumulation Spectrometry (ToF-SIMS) is performed in a ToF.SIMS5-100 (ION-TOF GmbH, Germany). Ultra-High Vacuum (UHV) abject burden is <5 × 10−9 mbar. For aerial accumulation resolution the Bi antecedent is operated in a bunched approach accouterment abbreviate ({{rm{Bi}}}_{3}^{ }) primary ion pulses at 25 keV activity and a crabbed resolution of approx. 4 µm. The abbreviate beating breadth of 1.5 ns allows aerial accumulation resolution. Charge advantage during spectrometry is all-important due to the awful careful attributes of the bottle substrates. Therefore, an electron flood gun accouterment electrons of 21 eV is applied, and the accessory ion reflectron acquainted accordingly. Images with a acreage of appearance of several aboveboard millimeters are acquired by scanning the sample date and are recorded with a 10 µm pixel size. For aerial crabbed resolution imaging at the approach edge, in adjustment to actuate the apery fidelity, a non-bunched primary ion approach is used. With nominal accumulation resolution the signals of SiO2, SiO3, and SiO3H are recorded with 256 × 256 pixel in a acreage of appearance of 500 µm × 500 µm. The arrangement allegiance at the approach bend is acquired by abacus the bottle and siloxane signals alongside to the edge.

Blocks of epoxide adhesive are able application a two-component epoxide embedding kit (EpoFix kit, Science Services, Germany) with a aggregate arrangement 15: 2 of adhesive to hardener and convalescent for 2 canicule at allowance temperature. The blocks are akin to a archetypal hardly trapezoidal appearance with 0.85 mm amplitude on the abbreviate ancillary and 1.4 mm acme application a 90° design accent knife (Trim 90, Diatome, Switzerland). The block is again cut into 60-nm attenuate sections application an ultramicrotome (Powertome XL, RMC, USA) and a adapted design knife (Ultra Jumbo, Diatome, Switzerland) with an added advanced baiter to acquiesce crabbed movement of the substrate beyond the complete breadth of the design knife. Sectioning is performed with accepted settings as recommended by the knife manufacturer. The substrates are blooming according to the subtractive action by plasma-induced siloxane abasement and abysmal into the DI water-filled backlog of the adapted design knife boat. Application a motorised 3-axis administration system, the substrates are again aerial to acquiesce dewetting in the berserk ascendancy areas and the accumulation of a attenuate baptize blur on the hydrophilic channels. Afterwards the channels of the arrangement are accumbent with the area award and added sectioning is performed to advance the area award central the channel. When a approach is abounding with sections, the area award is afar manually, and the substrate confused alongside to adjust the abutting abandoned approach with the end of the area ribbon. The action of cutting, break and substrate alive is again again to ample the actual channels with sections. Afterwards all channels are filled, the final abasement of the sections on the substrate is able by boring appropriation the substrate out of the baptize backlog and acceptance the actual baptize to air dry.

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