To observe actin filament organization in guard cells, Arabidopsis guard cells were imaged using a spinning disc confocal microscopy, collecting 0.5 μm interval serial sections, using an UPlanSApo 100x/1.40 objective lens on an Olympus IX70 and a CSU10 scanning head (Yokogawa Electric Corporation, Tokyo, Japan). To capture images of guard cell MFs, cotyledons were floated, with its abaxial surface up, on a glass slide covered with basal buffer [50 mM KCl, 5mM 2-(N-morpholine)-ethanesulphonic acid (MES)-Tris, pH 6.5, and 10 mM CaCl2], and then covered by a coverslip. For sample selection and analysis, a recent study by our group [39 ] demonstrates that both mature (ca. 4-6-week-old) plants and 11-14-day-old cotyledons show similar behavior and biological activity with elicited with biotic stressors (e.g., pathogens and purified pathogen elicitors)
Uplansapo 100x 1.40 objective lens
Manufactured by Olympus
Sourced in Japan
The UPlanSApo 100x/1.40 objective lens is a high-performance microscope objective designed for use in research and scientific applications. It features a magnification of 100x and a numerical aperture of 1.40, which allows for high-resolution imaging and excellent light-gathering capabilities. The lens is optimized for use with Olympus microscopes and is constructed with premium optical elements to provide clear, sharp images.
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2 protocols using uplansapo 100x 1.40 objective lens
1
Visualizing Actin Filaments in Arabidopsis Guard Cells
2
MDCK Cell Immunofluorescence Staining
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MDCK cells were fixed with 4% paraformaldehyde in PBS for 15 min at room temperature, followed by permeabilization with 0.2% Triton X-100 in PBS for 5 min. The samples were then incubated with 1% BSA in PBS for 1 h at room temperature, followed by sequential incubation with primary and secondary antibodies diluted with 1% BSA in PBS overnight at 4 C (primary antibodies) or for 1 h at room temperature (secondary antibodies). Images were collected using a Fluoview FV1000 confocal microscope (Olympus) equipped with a UPlanSApo 60x/1.35 or a UPlanSApo 100x/1.40 objective lens (Olympus).
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