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The Impact of the T Factor
The Turner Designs Technical Support Department strives to keep customers as informed as possible by constantly researching, updating, reviewing, and making resources available.  A good understanding of how temperature affects fluorescence is required for fluorometric analyses and we have references in our FAQ’s and application notes stating details and formulas regarding temperature corrections.  However, this is so critical to fluorescence measurements we thought we’d review it here.  In general, as temperature increases fluorescence decreases.

For some of our environmental customers, the temperature variance and subsequent effect on fluorescence is seasonal.   For some of our industrial customers that are using our instruments in boilers and cooling towers, the temperature variance is much greater.  In both cases, the difference between the temperature of the instrument during calibration and the temperature of the sample needs to be well-documented for correcting fluorescence data thereby maximizing accuracy of concentration estimates.  

For example, a two degree change in temperature for Rhodamine WT correlates to a 5.2% change in fluorescence due to temperature (2 x 0.026, expressed as a percentage).

Other examples for some common dyes:

Fluorescein
Temp Change Reading Drop
2 °C 0.72%
20 °C 7.20%
30 °C 10.80%
50 °C 18.00%
75 °C 27.00%
PTSA
Temp Change Reading Drop
2 °C 0.25%
20 °C 2.52%
30 °C 3.78%
50 °C 6.30%
75 °C 9.45%


Here are links to a few articles with more information about temperature’s affect on fluorescence:

A Temperature Compensation Method for CDOM Fluorescence Sensors in Freshwater

FAQ's: Fluorescence is temperature sensitive - coefficients and formula