fluorescein
Post on 21-Jul-2015
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Meet (or Re-Meet) Fluorescein
http://en.wikipedia.org/wiki/Fluorescein
http://en.wikipedia.org/wiki/Fluorescein
“The fluorescence of this molecule is very high, and excitation occurs at 494 nm and emission at 521.”
Absorption and Emission of Light
From: Molecular Probes
http://www.invitrogen.com/site/us/en/home/References/Molecular-Probes-The-Handbook/pH-Indicators/Probes-Useful-at-Near-Neutral-pH.html#head1
Obtain fluorescein sample from TA; blank with appropriate buffer & re-blankwhen you switch to a different pH. Determine appropriate dilution to use, and scan from 200 – 600 nm. Move cursors to obtain absorbance values at a particular lambda max.
Analyze your data using
(1) Nonlinear fitting
(2) Linear fitting
(3) 2nd Derivative fitting
To speed things up, we will read at 240 & 490 nmrather than scanning from 200 – 600 nm.
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