pDR195+ApDMT+EGFP, pDR195+ApDMTRP+EGFP, and empty vector pDR195 were introduced into competent yeast cells DY1457 using a frozen-EZ Yeast transformation II kit (Zymo Research Co., CA, USA). Transformants were selected on plates containing the synthetic dropout medium with appropriate amino acids. These yeasts were incubated in 1 mL of synthetic defined (SD)-Ura medium buffered at pH 6.0 with 50 mM 2-morpholinoethanesulfonic acid at 30 °C for 24 h and collected by centrifugation for 5 min at 6000 × g. Afterward, cells (OD600 = 0.1) were washed with sterile water and cultured in 1 mL SD-Ura medium (pH 6.0) at 30 °C overnight. A fluorescent microscope (Olympus BX-53 equipped with a high-pressure mercury lamp, U-HGLGPS, and a fluorescent filter cube, U-FGFP; Olympus, Tokyo, Japan) was used to observe expressed fusion proteins, and images were captured using a digital camera DS-Ri1 (Nikon, Tokyo, Japan).
U fgfp
Manufactured by Olympus
Sourced in Japan
The U-FGFP is a microscope objective lens designed for fluorescence imaging. It is optimized for use with green fluorescent protein (GFP) and other related fluorescent proteins. The U-FGFP provides high numerical aperture and transmission efficiency to enable efficient excitation and collection of fluorescence signals.
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Lab products found in correlation
2 protocols using u fgfp
1
Yeast Transformation and Fluorescent Protein Expression
2
Quantitative Autophagosome Morphometry
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Cells were cultured in SDCA medium to midlog phase, and autophagy was induced when necessary. Cells were harvested and subjected to fluorescence microscopy on an IX83 inverted system microscope (Olympus) equipped with a UPlanSApo100×/1.40 Oil (Olympus) and a CoolSNAP HQ CCD camera (Nippon Roper). A U-FGFP and U-FRFP filter sets (Olympus) were used for visualization of GFP/mNeonGreen and mRFP/FM 4-64, respectively. Images were acquired using the MetaVue imaging software (Molecular Devices). For determination of AM length, the fluorescence images of mNeonGreen-Atg8 were analyzed using Qautas (Kawaoka et al., 2017 (link)). FM 4-64 staining was performed as previously described (Suzuki et al., 2002 (link)). Data analysis was performed using R (https://www.r-project.org ). Boxplots were drawn using the default settings of R.
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