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Defining the Relationship Between Vacuole Size and pH
Starting a preliminary study in the cells ability to maintain homeostasis under pH stresses
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Background Info - The Lab I work in
❏ Dr. Mark Chan’s Lab, focuses on yeast (Saccharomyces cerevisiae)
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Background Info - The Lab I work in
❏ Larger goal is to find relationships
❏ Interested in size and shape of the vacuole
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Background - My Project and Its Place in Lab
❏ Continuation of former project involving pH
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Background - My Project and Its Place in Lab
❏ Results would give insight on acidity and size
❏ Next step was at the individual level
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Why Does This Matter? ❏ Yeast optimization and diagnosis❏ Results would give insight on acidity and size
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Goal of the Project - Two Objectives
1. Confirm a method to observe both size and acidity2. Utilize the method to gather data
❏ Majority of the summer was spent on goal #1
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So, what did I Use?
❏ A green, pH sensitive dye called Carboxyfluorescein Diacetate
❏ A red dye that binds to the vacuole surface, FM464
Moves into cell, becomes digested Changes fluorescence based on pH
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So, what did I Use?
❏ Media buffers at specific acidities
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Trials With Dye Uptake
Lack of brightness inside the cell
Presence of brightness inside the cell
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And, what did I do?
❏ collection of preliminary data
FM464 dye on the exterior of vacuoles. CFDA fluorescence inside cells.
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Expected Results
Acidic Vacuole Acidic VacuoleBasic VacuoleBasic Vacuole
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Expected Results
No Difference between the vacuoles
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Future of The Project
❏ Collection of more data and confirmation of results
❏ Expansion of the acidic ranges❏ Use of transformed yeast
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Acknowledgements
❏ Mark Chan and everyone from the lab
❏ SFSU Bridges Program
❏ Dr. Fredericks