User:Christian Niederauer/Notebook/Phgradients
Description
This Lab Notebook originally was written in another wiki -- this is the ported version. The project's title is Creating pH Gradients with Proton Uncaging Molecules. It was pursued as the experimental part of my bachelor's thesis.
Link: Lab Notebook
Thesis Abstract
This thesis approaches the photochemical creation and observation of pH gradients. The basics of acid-base chemistry and photochemistry are explained, with further detail dedicated to the photolysis of the used chemical species, 2-nitrobenzaldehyde (NBA). A fluorescence microscopy setup with an implemented ultraviolet laser and a synchronized chopper system is introduced and used for the photolysis of NBA and simultaneous observation of pH with a pH sensitive, ratiometric fluorescent dye (SNARF). The realization of ratiometric fluorescence analysis in the setup and the image processing is discussed. Characteristics, magnitude and shape of the created pH gradients, as well as the motion of charged particles in the gradient, are presented in the experimental section of the thesis.
Notebook
December 2013
18.12.13 SNARF & NBA in H[math]\displaystyle{ _2 }[/math]O without buffer
| x mM NBA | 0 | 1 | 2 | 4 | 8 |
| H[math]\displaystyle{ _2 }[/math]O in µl | 49 | 42.75 | 36.5 | 24 | 0 |
| NBA in µl | 0 | 6.25 | 12.5 | 25 | 49 |
| SNARF in µl | 1 | 1 | 1 | 1 | 1 |
SNARF stock concentration: 1mM
NBA stock concentration: 8 mM
Volume: 50µl
SNARF concentration: 20 µM (V/V_sample = c/c_stock)
