Laboratory Course in Tree Cell Biology2

Numbering Code G-AGR02 6BC39 EJ80 Year/Term 2022 ・ Year-round
Number of Credits 5 Course Type Experiment
Target Year Target Student
Language Japanese Day/Period
Instructor name SUGIYAMA JUNJI (Graduate School of Agriculture Professor)
YOSHINAGA ARATA (Graduate School of Agriculture Associate Professor)
AWANO TATSUYA (Graduate School of Agriculture Assistant Professor)
Outline and Purpose of the Course Newly developed fixation methods, such as high pressure freezing, are applied to differentiating xylem of angiosperm and gymnosperum to fix the cells in living state. Immunolabeling of hemicelluloses and lignin are applied to the differentiating xylem to observe their deposition and distribution.
Course Goals ?Develop the skills to conduct research independently by mastering a wide range of experimental methods for investigating the lignified cell wall;
?Develop the skills to conduct research independently by mastering a wide range of experimental methods for investigating the cell wall during its formation.
Schedule and Contents 3-5 weeks of class are planned for each of the topics covered in this course, including the following:
1. Electron microscopy
  Students will enhance their understanding of the basic principles and applications of methods for investigating ultrastructures using transmission and scanning electron microscopes.
2. Image processing
  Students will enhance their understanding of the techniques for conducting two-dimensional and three-dimensional analyses of cell morphology by using a standard computerized image processing method and Fourier Transform.
3. Diffraction methods
  Students will enhance their understanding of the common principles of X-ray diffraction, electron diffraction, and optical diffraction and their individual characteristics and practical applications.
4. Observation methods for cambium activity
  Students will enhance their understanding of the methods of collecting and fabricating samples for tracking the processes of cell division and production and xylem differentiation in trees, primarily by using optical microscopes.
5. Immunohistochemical tracking methods
  Students will enhance their understanding of the basics and applications of immunohistochemical staining, a method used to visualize the locations of cell wall components and enzymes related to their synthesis.
6. Localization of cell wall components
  Students will enhance their understanding of the principles of methods such as microspectrophotometry, fluorescent photometry, and component analysis methods for extremely small samples, and problems associated with the application of such methods.
7. Freezing and observation methods for storing ultrastructures in their living state
  Students will enhance their understanding of the principles and techniques of rapid-freeze methods and freeze-replica methods to store a variety of cell organelles in good condition.
Evaluation Methods and Policy Evaluation will be based on attendance.
Refer to '2017 Guide to Degree Programs' for attainment levels of evaluation.
Course Requirements Background knowledge: optics, X-ray diffraction, biochemistry, botany, and cytology.
Study outside of Class (preparation and review) Conduct your experiments while staying in close communication with the supervisor.
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