Plants and Photosynthesis. Plant cell Only in Plants Chloroplast Cell Wall Vacuole (?)
Defining the Relationship Between Vacuole Size and pH (1)
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Transcript of Defining the Relationship Between Vacuole Size and pH (1)
Defining the Relationship Between Vacuole Size and pH
Starting a preliminary study in the cells ability to maintain homeostasis under pH stresses
Background Info - The Lab I work in
❏ Dr. Mark Chan’s Lab, focuses on yeast (Saccharomyces cerevisiae)
Background Info - The Lab I work in
❏ Larger goal is to find relationships
❏ Interested in size and shape of the vacuole
Background - My Project and Its Place in Lab
❏ Continuation of former project involving pH
Background - My Project and Its Place in Lab
❏ Results would give insight on acidity and size
❏ Next step was at the individual level
Why Does This Matter? ❏ Yeast optimization and diagnosis❏ Results would give insight on acidity and size
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
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
So, what did I Use?
❏ Media buffers at specific acidities
Trials With Dye Uptake
Lack of brightness inside the cell
Presence of brightness inside the cell
And, what did I do?
❏ collection of preliminary data
FM464 dye on the exterior of vacuoles. CFDA fluorescence inside cells.
Expected Results
Acidic Vacuole Acidic VacuoleBasic VacuoleBasic Vacuole
Expected Results
No Difference between the vacuoles
Future of The Project
❏ Collection of more data and confirmation of results
❏ Expansion of the acidic ranges❏ Use of transformed yeast
Acknowledgements
❏ Mark Chan and everyone from the lab
❏ SFSU Bridges Program
❏ Dr. Fredericks