1. Functional Analysis of Ionic Flux in Isolated Endosomes
Michael Carrithers
Department of Neurology

2. Can voltage-gated ionic flux enhance acidification of macrophage endosomes during infection?

3. Goal: Use flow cytometry on isolated endosomes to complement microscopy and electrophysiology.

4. Experimental Approach
Differentiate and prime a monocytic cell line (THP-1).
Isolate endosomes on a sucrose gradient.
Dialyze sample.
Load with functional fluorescent dyes.
Activate channel.
Analyze on MoFlo.
Collect enriched endosomal fraction.

5. Fluorescent Dyes Lysosensor Green (pH) Sodium Green CoroNa Red
Disbac2(3) (slow voltage-sensitive dye)

6. Technical Challenges Particle size (200-800 nm) Pressure Sheath buffer
Large amount of starting material