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Application of Environmental Scanning Electron Microscope-Nanomanipulation System on Spheroplast Yeast Cells Surface
Maryam Alsadat Rad1, Mohd Ridzuan Ahmad1, Masahiro Nakajima2
1Department of Control and Mechatronic Engineering, Faculty of Electrical Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia.
Scanning
|November 8, 2017
Summary
Researchers successfully converted W303 yeast cells into spheroplasts, confirming the morphological changes using microscopy and Environmental Scanning Electron Microscopy (ESEM). Optimal conversion was observed in specific buffer solutions within 15 minutes.
Area of Science:
- Cell Biology
- Microscopy Techniques
Background:
- W303 yeast cells possess a rigid cell wall essential for their structure.
- Spheroplasts are cells with their cell walls removed, altering their morphology and properties.
Purpose of the Study:
- To prepare and characterize spheroplasts from W303 yeast cells.
- To confirm the spheroplasting process using various microscopy and quantitative methods.
- To evaluate the effect of different media on spheroplast formation.
Main Methods:
- Spheroplast preparation from W303 yeast cells.
- Optical microscopy for morphological evaluation.
- Spectrophotometry (OD800) for quantitative conversion assessment.
- Environmental Scanning Electron Microscopy (ESEM) for surface characterization.
Main Results:
- Successful qualitative and quantitative confirmation of W303 cell spheroplasting.
- Normal cells are oval; spheroplasts are spherical.
- Sorbitol-EDTA-sodium citrate buffer (SCE) and sorbitol-Tris-Hcl-CaCl2 (CaS) media showed higher spheroplast yields.
- Complete conversion achieved within approximately 15 minutes.
- ESEM revealed smooth surfaces on normal cells and bleb-like surfaces on spheroplasts.
Conclusions:
- The study successfully demonstrates the preparation and characterization of W303 yeast spheroplasts.
- Specific buffer solutions enhance spheroplast formation efficiency.
- ESEM provides detailed insights into the surface changes associated with cell wall removal.
