Preparing samples with optimal cutting temperature compound and observing at −30☌ helped to increase the stability of Coolstage samples.
Transmission light microscope images series#
The use of methanol and a graded series of steps improved the preservation of samples. The results indicate that methanol fixative obviously reduced shrinkage of SEM samples compared with glutaraldehyde and formaldehyde alcohol acetic acid (FAA) fixatives. The dimension and morphology of tissues were compared using the six different methods. Thus, we compared four different t-butanol freeze-drying methods and two Coolstage methods. In addition, fresh samples were not stable on the Coolstage. Moreover, we usually used glutaraldehyde as an SEM fixative when using t-butanol for freeze-drying, though methanol is considered a better fixative in recent studies. However, stomatal apparatuses of Arabidopsis, cucumber, radish, and maize deformed and showed poor appearance when using the Coolstage. The resolution of images of conventional SEM leaf samples was generally higher than the Coolstage images at the same magnification, whereas local collapse and shrinkage were observed in leaves with high water content when using the conventional method. Investigation of samples cleared by chloral hydrate is convenient and autofluorescence of cell walls can be detected in rice. Different parts of epidermis peels were well focused under the LM. SEM images were obtained using a standard stage for conventional dehydrated samples or a Coolstage for fresh tissue. Optical microscope specimens were obtained using either the direct isolation method or the chloral hydrate-based clearing method. Here, four specimen preparation methods were used to investigate the leaf epidermis morphology of Arabidopsis, radish, cucumber, wheat, rice, and maize, under an inverted basic light microscope (LM), a laser scanning confocal microscope (LSCM), or a scanning electron microscope (SEM). There have been few studies comparing different preparation and imaging methods for visualizing the leaf epidermis. The micro-morphology of leaf epidermises is valuable for the study of leaf development and function, as well as the classification of plant species. Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs, College of Life Sciences, Henan Normal University, Xinxiang, China.Jinhong Yuan Xiaoduan Wang Huihui Zhou Yulin Li Jing Zhang Shuxin Yu Mengni Wang Menghan Hao Qian Zhao Le Liu Mingjun Li * Junhua Li *
0 kommentar(er)
