15 rhodamine wt, Introduction and principle of operation – Xylem 6-Series Multiparameter User Manual

Page 266

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Principles of Operation

Section 5

YSI Incorporated

Environmental Monitoring Systems Manual

5-28

content of an environmental sample may appear to the eye to be relatively stable, the displayed

chlorophyll reading can vary significantly depending on the nature of the particles in the optical path at

the instant of measurement. In a discrete sample study of environmental water, for example, the

variability of the output can be significant. This apparent jumpiness is not observed in dye standards,

since these are homogeneous solutions containing no suspended matter.

The sonde chlorophyll system allows the user to apply a mathematical filter to the raw data so that the

sensor output may be more representative of the average phytoplankton content of the environmental

sample. From the Advanced|Sensor menu of the sonde software, the user can activate the data filter

and adjust its performance. For typical sampling and monitoring applications, YSI recommends that

the Data Filter settings for chlorophyll be selected as follows: Enabled -- On; Wait for Filter -- Off;

Chlorophyll Time Constant = 12; Chlorophyll Threshold = 1. The advantage of the filter is a more

stable display of chlorophyll readings.

5.15

RHODAMINE WT

INTRODUCTION AND PRINCIPLE OF OPERATION

Rhodamine WT is a red dye that is commonly used in stream flow studies. The amount of the species at

various points (horizontal and vertical) in the water under examination is determined by utilizing the fact

that Rhodamine WT fluoresces when irradiated with the proper wavelength of light and thus the YSI 6130

Rhodamine WT sensor works on the same principles as described above for the 6025 chlorophyll sensor.

As for chlorophyll, Rhodamine WT fluoresces, that is, when irradiated with light of a particular

wavelength, it emits light of a higher wavelength (or lower energy). The ability of Rhodamine WT to

fluoresce is the basis for all commercial fluorometers capable of measuring the analyte in situ.

Fluorometers of this type have been in use for some time. These instruments induce Rhodamine WT to

fluoresce by shining a beam of light of the proper wavelength into the sample, and then measuring the

higher wavelength light which is emitted as a result of the fluorescence process. Most Rhodamine systems

use a light emitting diode (LED) as the source of the irradiating light that has a peak wavelength of

approximately 540 nm. LEDs with this specification produce radiation in the visible region of the spectrum

with the light appearing green to the eye. On irradiation with this green light, Rhodamine WT in the water

emits visible light with a higher wavelength than that of the exciting beam, i.e. the Rhodamine fluoresces.

To quantify this fluorescence, the system detector is usually a photodiode of high sensitivity that is

screened by an optical filter that restricts the detected light. The filter minimizes (a) the exciting light being

detected when it is backscattered off of particles in the water and (b) the interference from other fluorescent

species such the chlorophyll in phytoplankton. Without the filter, turbid (cloudy) water or water with high

levels of phytoplankton would appear to contain Rhodamine WT, even though none were present. The

following diagram can be used to better understand the principles of the YSI Rhodamine WT system.

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