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The motorized stage was fixed to a three-axis manual rotation stage mounted on a photography tripod.
Afterward, the sample stage was fixed at point C. The force curves measured after fixing the sample stage are shown in Fig. 2b.
Then, the measured force returned to 0 N, the sample stage was fixed, and the changes in detected force with time were recorded.
The stage was fixed on a CLSM confocal microscope (LSM 510, Carl Zeiss MicroImaging GmbH, Germany).
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Isolated oocytes at the GV stage were fixed in PHEM fixative, and indirect immunocytochemistry of the oocytes was performed as previously described (Fernandes et al., 2012).
In addition, the general mean and variance of quality characteristic in the third stage are fixed for each observation.
In order to conduct SEM and EDX analysis of NO/THCPSi NPs treated and untreated with E. coli, colonies at the desired growth stage were fixed by formaldehyde (4 v/v%) for 2 h on round graphite disks.
Anthers at the meiotic stage, the early and late uninucleate microspore stages, the bicellular pollen stage, and the tricellular pollen stage were fixed with a formaldehyde-acetic acid-ethanol solution, and embedded in Technovit 7100 resin (Kluzer, Wehrheim, Germany) as described by Zhou et al. (2011).
Embryos at the 38 somite stage were fixed overnight in 4% PBS (Electron microscopy services) overnight at 4°C.
To label filamentous actin (F-actin), zebrafish embryos at designated stage were fixed in 4% paraformaldehyde in PBS overnight at 4°C, washed in PBS and manually dechorionated.
Embryos collected at the tail-bud stage were fixed overnight in 4% parafolmaldehyde/PBS at 4°C, were hand-dechorionated and dehydrated overnight in methanol at −20°C.
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