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