Limitations of accuracy for the bga-pe sensor, Estimation of the bga-pe range in cells/ml – Xylem 6-Series Multiparameter User Manual

Page 280

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Principles of Operation

Section 5

YSI Incorporated

Environmental Monitoring Systems Manual

5-42

mechanical wiper that makes it ideal for unattended applications. The wiper can be activated in real-time

during discrete sampling operations or will function automatically just before each sample is taken during

long term unattended monitoring studies. The number of wiper movements and the frequency of the

cleaning cycle for the unattended mode can be set in the sonde firmware. Generally, one wiper movement

is sufficient for most environmental applications, but in media with particularly heavy fouling, additional

cleaning cycles may be necessary.

LIMITATIONS OF ACCURACY FOR THE BGA-PE SENSOR

As mentioned above, the measurement of PE-containing BGA from in vivo fluorescence measurements will

almost always be less accurate than determinations made using either cell counting or spectrofluorometric

quantitation of molecular PE after its extraction from cells. Some of the reasons for this accuracy

limitation with in vivo BGA-PE measurement include the following:

Interferences from other microbiological species such as chlorophyll-containing phytoplankton

Interference from sample turbidity.

Differences in the general fluorescence intensity of different PE-containing BGA species

Differences in the effect of temperature on the fluorescence intensity of different PE-containing

BGA species

Effect of the variation in ambient light conditions on BGA fluorescence and differences in this

effect between different PE-containing BGA species.

In addition, when present in high concentrations, colonies of BGA can often be seen with the naked eye

and may resemble fine grass cutting or take the form of small irregular clumps or pinhead-sized spheres.

When BGA colonize into these forms, the sensitivity of the YSI sensor in terms of the fluorescence per cell

of BGA is reduced because it has been designed to detect microscopic, free-floating cells and not large,

macroscopic floating particles. Thus, the sensor is likely to underestimate the total amount of BGA

present in the water when clumps are present.

Users should take careful note that these limitations mean that any in vivo sensors such as BGA-PE and

chlorophyll will be significantly less quantitative than any of the other sensors offered for use with YSI

sondes and make it impossible for YSI to provide an actual accuracy specification in cells/mL for the 6132

PE Probe.

ESTIMATION OF THE BGA-PE RANGE IN CELLS/ML

As noted in the above section, the use of in vivo phycoerythrin fluorescence to estimate the cell content of

PE-containing algae has significant limitations. These limitations also make the designation of a range (or

full scale sensor reading) for any BGA-PE sensor less than quantitative. The range estimate for the YSI

6132 sensor is based on the fact that its reading in an empirical sample of PE-containing algae is very

similar to that of the industry standard fluorometer from Turner Designs which is configured for BGA-PE.

In the estimation experiment, the Turner Cyclops was fixed on its middle range and then its voltage reading

in a BGA-PE culture (a Synechococcus sp.) was determined. The YSI 6132 sensor was placed in the same

culture and its sensitivity adjusted to show the same percent of full scale deflection as the Turner sensor on

its middle range. Since Turner Designs has designated the middle range of its sensor as 0-200,000

cells/mL, the YSI sensor is estimated to have the same range. Naturally, this range is only an estimation

for both the YSI and Turner sensors because of the general limitations of in vivo fluorescence

measurements described above.

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