Bactiquant Photomanual Manual filtration 2021

PREPARE CALIBRATION

2021

PHOTO MANUAL FOR ® BACTIQUANT WATER 3. Use a black cuvette and a standard

- Manual Filtration

4. Press ”CAL” 5. Press ”ENT” 6. Place the black cuvette in the fluorometer and press ”ENT”

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PREPARE CALIBRATION Make sure all chemicals are equilibrated to room temperature 1.

Turn on the fluorometer Press ”ON” Display should read UV and 0,0

2. Press

”STD VAL” on the fluorometer Make sure the value is the same as listed on the reverse side of the fluorometer

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PREPARE CALIBRATION 3. Use a black cuvette and a calibration standard

4. Press ”CAL” 5. Press ”ENT” 6. Place the black cuvette in the fluorometer and press ”ENT”

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CALIBRATION 7. The calibration standard liquid is transferred to the cuvette. Avoid touching the cuvette on the lower part 8. Remove air bubbles by gently tapping the cuvette with a finger nail

9. Place the cuvette in the fluorometer. Close the lid and press ”ENT” 10. Press ”ENT” to finalize the calibration

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CALIBRATION 11. Leave the cuvette in the fluorometer and press ”READ” The value should be within 3 % of the standard value This value is termed ”Standard Value Measured”

12. Place the black cuvette in the fluorometer and press ”READ”

The value should be between 0 +/- 1

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CALIBRATION 13. Note the ”Standard Value” on the analysis sheet, using the BQ APP or directly in the spreadsheet

You have now calibrated the fluorometer

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ANALYTICAL BLANK 1. Place a cuvette with a developer tube in the outermost right square of the rack. Transfer the devloper in to the cuvette

2. Attach the syringe to the substrate bottle 3. Push the syringe and the syringe mounting platform all the way down. There is a click

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ANALYTICAL BLANK 4. Turn the substrate bottle up side down. Transfer 0.35 mL substrate to the syringe.

5. Transfer the substrate to the developer in the cuvette

Mix by pumping the liquid up and down three times in the cuvette using the syringe. Remove air bubbles in the cuvette

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ANALYTICAL BLANK 6. Place the cuvette in the fluorometer and close the lid.

7. Press ”READ” This value is termed the ”Blank Value”

8. Note the blank value in the analysis sheet, APP or directly in the spreadsheet

The ”Blank Value” should be less than 200

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SAMPLE FILTRATION Items needed for sample filtration include: • Sterile unused 0.2 µm filters • Sterile unused 60 ml syringes • Water samples • Marker 1.

Record sample date and sample ID on the analysis sheet, APP or directly in the Excel spreadsheet

2. Mark the sample ID on the filter

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The filtration step is performed to concentrate the bacteria in the water sample onto the filter for increased sensitivity.

SAMPLE FILTRATION 3. Transfer water sample to be tested to the new, unused 60ml sampling syringe and record the sample volume on the analysis sheet

4. Attach the sampling syringe with the water sample to the new unused filter disc marked with the corresponding sample ID. Rotate the filter disc until it fits snugly on the syringe.

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SAMPLE FILTRATION 5. Push the water sample through the filter and ensure that excess sample (filtrate) is collected into a proper waste container.

6. Evacuate the dead volume in the filter by detaching the syringe from the filter. Fill it with air, reattach the syringe and use the air to push out the remaining liquid into a proper waste container.

7. Discard syringe.

Note: If sample has a high particulate content, it may not be possible to filter 60mls. Always record actual volume filtered.

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PREPARE THE ANALYSIS Retrieve substrate, filters, blunt needles, cuvettes with developer and syringes, according to the number of samples to be analysed Place the items in the rack as shown on the picture to the right

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PREPARE THE ANALYSIS 1. Transfer 2,5 mL substrate to the syringe

2. Gently attach the syringes on the filters.

Then place the syringes with filters on the rack as shown on the picture 3. Empty the developer into the cuvettes

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START THE ANALYSIS 4. Flush the filter with substrate 5. Surplus liquid is collected in the glass vial for disposal 6. Attach the filter with syringe to the blunt needle

7. Rapidly repeat the routine for each sample 8. Start the stopwatch or timer in the APP

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TERMINATION OF THE ANALYSIS 9. After 15 minuttes the assay is terminated by placing the filter with the syringe and blunt needle in the developer and back flushing two times 10. Rapidly repeat the routine for the remaining samples

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TERMINATION OF THE ANALYSIS 11. Detach the syringe from the filter. Fill it with air reattach the syringe and use the air to push out the remaining liquid in the filter into the cuvette

12. Turn on the fluorometer 13. Gently remove air bubbles

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TERMINATION OF THE ANALYSIS 14. Place the cuvette in the fluorometer. Close the lid 15. Press ”READ”

16. Note the fluorescence reading (analysis value) on the analysis sheet, APP or directly in the spreadsheet 17. Use the APP or spread­ sheet to calculate the BQ value. spreadsheet can also be used as a tool for calculating baseline values

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NOTES

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IF YOU NEED ASSISTANCE OR AN EXTRA HAND, CALL BACTIQUANT TO BE CONNECTED TO ONE OF OUR PRODUCT SPECIALISTS info@bactiquant.com +45 6988 4000

www.bactiquant.com

STORAGE INSTRUCTIONS The bottles of Standard solution, Developer solution and Substrate solution should be stored in a refrigerated location at approximately 4°C until ready for use. All solutions must be at room temperature, prior to analysis.

Dry goods (including syringes, blunt needles and filters) should be stored at room temperature in a clean, dry location.

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TROUBLESHOOTING BACTIQUANT WATER (INCLUDING DISPENSER ASSAYS)

ANALYTICAL BLANK values >200 - values greater than 200 indicate the substrate has deteriorated and is no longer valid for analysis. This can occur if it is not properly stored in the refrigerator, if it has been exposed to UV light for prolonged periods of time or if it has exceeded its shelf life. The substrate solution must be discarded if the analytical blank value is above 200.

How often is a new analytical blank needed? A new analytical blank must be made for each bottle of substrate used during the analysis of samples. If several sets of samples are analyzed sequentially, but within one hour of time, The same analytical blank value can be used, provided the same substrate bottle is used for all samples analyzed.

Sample analysis value reads “over” – if the fluorometer reads “over” it is indicating the fluorescent value for that sample is in excess of the instrument range.

In order to bring the sample back into range, a simple dilution can be made by extracting 100 µl of the sample substrate/developer solution in the cuvette and adding it to a clean, new cuvette. Add 2ml of developer to the 100 µl and stir well. Reread the new sample solution. Record the new value in the spreadsheet and change the multiply factor column to 21. Use the same, temperature, reaction time, sample volume and blank value. The spreadsheet will recalculate the Bactiquant value for you.

Air in dispenser ( how long can it sit before you have to reprime it) Prime the dispenser before each set of analysis by discarding the first pump. Refer to the instructions in this manual.

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Negative values can occur when the result of the Bactiquant analysis is close to zero. The variation on zero is approximately +/- 10 FLU. Therefore negative values of up to -10 can occur. Negative values in excess of 10 FLU may be associated with cuvette errors such as bubbles, fingerprints, or scratches on the cuvette, particulate floating in the cuvette solution, a highly colored final solution in the cuvette or if the analytical blank value was not prepared correctly.

If a negative value in excess of 10 FLU occurs, and there are floating particles or a dark color due to micro particles in the cuvette solution, filter the cuvette solution and rerun the sample Two errors can occur when preparing the analytical blank. The first is in measuring the 0.35ml of substrate in the syringe. Care must be taken to precisely measure 0.35mls in the syringe. An inaccurate volume in the syringe can affect the analytical blank value and thereby influence the final result. The other error is due to insufficient mixing.

Cuvette errors – prior to using or reading solutions in a cuvette, they should be checked for bubbles, fingerprints, moisture or scratches. Remove bubbles; clean the outer surfaces of the cuvette with a lint free soft cloth prior to analysis. Scratched cuvettes should be discarded and not used for analysis. Only use cuvettes provided by Mycometer for analysis.

Spills in fluorometer chamber will not generally damage the chamber if they are soaked up immediately as it is watertight. Use a soft dry lint free cloth to soak up the liquid and thoroughly dry the interior of the chamber.

If the spill has not been soaked up immediately, use clean 23

TROUBLESHOOTING BACTIQUANT WATER (INCLUDING DISPENSER ASSAYS)

water to gently rinse the cuvette holder after the spill has been removed and thoroughly dry the chamber with a soft lint free cloth.

Battery changes the fluorometer uses 4 AAA batteries for operation. The battery chamber is located on the back of the instrument and requires a screwdriver to remove the o-ring sealed tight fitting cover. In general, batteries should be changed once a year.

Sample storage samples are best analyzed on the day they are collected. However, they can be held under refrigeration ( 4C) up to 24 hours.

Temperature range for the fluorometer and the analytical chemistry during analysis should be between 18-35C. The temperature of the samples collected is not important since the samples are filtered.

REACTION TIME Minimum reaction time is 10 minutes.

Run over if the time for analysis exceeds the time originally set, simply enter the actual time into the spreadsheet CHEMICAL SAFETY None of the chemicals in the BactiQuant kit are categorized as hazardous. For details on handling of the chemicals and first aid measures, refer to the Material Safety Data Sheets that are packed with each set of chemistry. The developer solution is alkaline. In case of eye contact rinse immediately with water for at least five minutes.

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TESTING FOR BACKGROUND FLUORESCENCE If there is a question as to whether the results of analysis are influenced by a background level of fluorescence not associated with microbial contamination, prepare a solution containing the chemical in the concentration in which it occurs in the product or liquid solution. Pour developer vial into a cuvette or if using a dispenser, dispense 2ml developer to a clean new cuvette. Measure the fluorescence of the developer in the fluorometer and record this value. Add 100 µl of the solution containing the chemical to the developer in the cuvette, mix well and measure again in the fluorometer. If the measured value of the chemical/developer is close to the developer only, the chemical does not fluoresce. Be aware that there are no particles in the sample. Particles in the solution will give turbidity and lead to a signal on the fluorometer. If there are particles the solution should be filtered before reading the fluorescence.

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PHOTO GLOSSARY

Cuvette

Assays components in alufoil bags, Blunt needle, Syringe (1.0 mL), Syringe (2.5 mL), Filter (0.2 µm),

Black Calibration cuvette

Developer Solution bottle

Expiration date on bag

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Dispenser

Substrate solution bottle

Standard solution

Fluorometer

Analysis Rack

Analysis sheet

Thermometer

Excel spreadsheet

Large 60 ml syringe

Timer 27

info@bactiquant.com · www.bactiquant.com