Measurement of chlorophyll, Optical fiber, Photodetector – Xylem 6-Series Multiparameter User Manual

Page 260: Light source, Blue led, Optical filter

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Principles of Operation

Section 5

YSI Incorporated

Environmental Monitoring Systems Manual

5-22

chlorophyll system are designed to complement the more accurate (but more difficult to obtain) results

from more traditional methods of chlorophyll determination.

MEASUREMENT OF CHLOROPHYLL

IN VIVO

One key characteristic of chlorophyll is that it fluoresces, that is, when irradiated with light of a particular

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

is the basis for all commercial fluorometers capable of measuring the analyte in vivo. Fluorometers of this

type have been in use for some time. These instruments induce chlorophyll 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 chlorophyll systems use a light emitting diode (LED)

as the source of the irradiating light that has a peak wavelength of approximately 470 nm. LEDs with this

specification produce radiation in the visible region of the spectrum with the light appearing blue to the

eye. On irradiation with this blue light, chlorophyll resident in whole cells emits light in the 650-700 nm

region of the spectrum. To quantify the 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 prevents the 470 nm

exciting light from being detected when it is backscattered off of particles in the water. Without the filter,

turbid (cloudy) water would appear to contain fluorescent phytoplankton, even though none were present.

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

Most commercial fluorometers fit into two categories. In the first category are benchtop instruments that

generally have superior optical flexibility and capability but are relatively expensive and are often difficult

to use in the field. In the second category are sonde-type fluorometers that have a fixed optical

configuration, but are less expensive, can be more easily used in the field, and are usually compatible with

data collection platforms. The use of a pump is recommended for some sonde fluorometers and this can

result in the need for large capacity batteries for field use.

The unique YSI chlorophyll system available as an option for use with YSI sondes consists of a probe

which is similar in concept to the sonde-type fluorometers, but is much smaller, making it compatible with

the probe ports of the YSI 6820V2-1, 6820V2-2, 6920V2-1, 6920V2-2, 600 OMSV2-1, 6600V2-2,

6600EDS V2-2, and 6600V2-4 sondes. The output of the sensor is automatically processed via the sonde

software to provide readings in either relative fluorescence units (Chl RFU) or µg/L of chlorophyll. No

pump is required for the YSI system allowing the sensor to operate off of either the sonde internal batteries

or the batteries in the YSI 650 MDS display/logger. Like the YSI 6026 and 6136 turbidity probes, the YSI

Optical Fiber

Photodetector

Light Source

Blue LED

Optical Filter

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