X cite 120q

ProtComp 9.0 (http://linux1.softberry.com/berry.phtml (accessed on 25 May 2021)) was employed for subcellular location prediction, and TMHMM (http://www.cbs.dtu.dk/services/TMHMM/ (accessed on 25 May 2021)) was utilized for protein transmembrane analysis. For gene subcellular localization, pJIT-166 was used as the target vector, and XbaI and BamHI were employed to generate restriction sites to construct a fusion expression vector. The primers used are shown in Supplementary Table S9. The expression vector was transfected into onion epidermal cells by microprojectile bombardment and incubated for 24 h in the dark [16 ]. The cells were subsequently observed using a fluorescence microscope (X-cite 120Q, Carl Zeiss).

The NtSOS2 open reading frame (ORF) was fused in-frame upstream to GFP in pJIT166 vector using XbaI and BamHI restriction sites. The primers used to construct this vector were listed in Supplementary Table 3. Then the constructive fusion plasmid was transfected into onion epidermal cells by microprojectile bombardment (Nebenführ, 2014 (link)) and incubated for 24 h in the dark. The cells were subsequently observed using Zeiss fluorescence microscope (X-cite 120Q, Carl Zeiss).

An optical contact angle measurement tool (DSA25, Krüss GmbH, Hamburg, Germany) is used to determine the equilibrium contact angles of water on the chip surface, which is obtained by averaging five measurement results at different locations on each chip. The experimental setup is shown in Figure 4. Optical inspections of the fluid flow in the microvalve structures are performed using an Axio Imager M2p (Carl Zeiss Microscopy GmbH, Jena, Germany) upright fluorescent microscope (see Figure 4a) equipped with a 20× objective with numerical aperture of 0.4 (Zeiss Objective LD Plan-Neofluar 20×/0.4 Corr M27, 421350-9971-000). The microscope is configured with a mercury vapor short arc lamp (X-Cite 120Q, Carl Zeiss Microscopy GmbH, Jena, Germany) and appropriate fluorescent filter sets for illumination and signal detection. Image acquisition is performed using a CCD camera (Axiocam506 mono, Carl Zeiss Microscopy GmbH, Jena, Germany). The chips are tested by using deionized water mixed with a fluorescence dye, Rhodamine B (Sigma Aldrich, St. Louis, MO, USA), at a concentration of 100 µg/mL. The mixture has a contact angle similar to pure water. The silicon chip is covered by a thin PDMS film to enclose the fluidic channels. The fluid situation in the microfluidic chip can be clearly observed using the Rhodamine B solution (as shown in Figure 4b).