Quorum cryo stage and transfer station

Cryo-SEM using a plunge freezing method was used for imaging and the samples were stored in liquid nitrogen [28 (link)]. Cryo-SEM visualization of samples was performed with FEI Versa3D equipped with a Quorum cryo stage and transfer station (FEI Company).

The VRSA cells were incubated with PBDM–5(6)–Carboxyfluorescein conjugates (15 and 10 μg/mL for PBDM1–5(6)–Carboxyfluorescein and PBDM2–5(6)–Carboxyfluorescein, respectively) for 30 min in dark and visualized under an inverted Olympus IX 71S8F-3 fluorescence microscope as discussed in the previous section. The images were processed using Stream Basic 1.7 Software (Milosavljevic et al., 2016 (link)).
Whereas, for Cryo-SEM the VRSA incubated for 4 h with PBDM peptides (15 and 10 μg/mL for PBDM1 and PBDM2, respectively) at 37°C in a shaking incubator and the control was VRSA without any treatment. Then the Cryo-SEM experiment method of plunge freezing was used. For plunging and storing of samples liquid nitrogen was used. Cryo-SEM visualization of samples was performed with FEI Versa3D equipped with a Quorum Cryo stage and transfer station (FEI Company) (Wu et al., 2014 ).