Bugcount® XLPD

Key terms

• ATP: Adenosine triphosphate, energy molecule of living cells.
• cATP (Cellular ATP): ATP inside living cells, representing active biomass.
• RLU: Relative Light Units (measurement from luminometer).
• ATP1 RLU: Calibration signal from UltraCheck 1.
• cATP RLU: Sample result for cellular ATP.

1.1 Log in to Relay™

• Log in (or create your account) at http://relay.luminultra.com.

If you are new to Relay™, follow the guides to first create a site and sample location.

• Connect to your luminometer, then select RUN TESTS to begin a new test.
• As you work through these test kit instructions, you can initiate RLU readings directly within Relay™ by selecting READ on any input. Once all inputs are entered, your results will automatically calculate.

1.2 Rehydrate the Luminase^XL enzyme

• Gently mix the Buffer and Luminase^XL enzyme.
• Wait 5 minutes for solution to dissolve.

After rehydration, keep Luminase refrigerated or frozen; it stays effective for 2–4 weeks at 4°C (39°F) or 3–6 months when frozen between uses. Before testing, let it thaw and reach room temperature (takes about 1 hour). Do not apply direct heat to warm it faster. For more details, visit my.luminultra.com.

1.3 Calibration using UltraCheck 1 (ATP1 RLU)

• Hold the UltraCheck 1 bottle vertical and add 2 drops (100µL) of it to a 12x55mm test tube.
• Pipet 100µL of Luminase^XL into the tube.
• Swirl the tube, place it in the luminometer, and take the ATP1 RLU reading (select READ) within 10 seconds.

If ATP1 RLU ≤ 50,000 rehydrate a new bottle of Luminase^XL.

Once UltraCheck 1 is opened, it must be used within 3 months. After 3 months, discard and use a new bottle.

2. Cellular ATP analysis (cATP RLU)

2.1 Pre-dilution

• Mix sample well.
• Add 1mL of sample to a 9mL UltraLute (dilution) tube.

• Remove the plunger from a new 20mL syringe and attach a filter.
• Pour the contents of the pre-dilution tube into the syringe.

• Slowly push sample through the filter into a waste receptacle.
• Detach the filter and remove the plunger.

2.2 Washing and drying

• Re-attach the filter to the 20mL syringe barrel.
• Using a disposable bulb pipet, add 5mL of LumiClean to the syringe barrel.
• Push the LumiClean slowly through the filter into a waste container.

• Remove the plunger from a 50mL syringe.
• Detach the filter from the 20mL syringe.
• Attach it to the 50mL syringe.
• Hold the 50mL syringe over the waste receptacle and push the plunger through the barrel to dry the filter.

Replace the 60mL syringe after every 20 tests.

2.3 Extraction

• Remove the plunger from the 20mL syringe.
• Remove the filter from the 60mL syringe and re-attach it to the 20mL syringe.
• Add 1 mL of UltraLyse 7 to the barrel using the 1mL pipettor.
• Pass it slowly through the filter and collect in a new 9mL UltraLute (dilution) tube.
• Cap and invert three times to mix.

2.4 Assay

• Add 100µL of the UltraLute (dilution) solution to a 12x55mm test tube.
• Use a new pipet tip to add 100µL of Luminase^XL.
• Swirl the tube, place it in the luminometer, and take the cATP RLU reading (select READ) within 10 seconds.
• In Relay™, select SAVE TEST.

3. Analysis

Tip: Relay™ does the work For you

When all inputs are entered in Relay’s Run tests workflow, the results will calculate automatically. Select Save Test to catalog and share your results with your team.

Calculations

\mathrm{cATP}\;(\mathrm{pg\ ATP}/\mathrm{mL})
=
\frac{\mathrm{cATP}_{\mathrm{RLU}}}{\mathrm{ATP1}_{\mathrm{RLU}}}
\times
\frac{100{,}000\;(\mathrm{pg\ ATP})}{V_{\mathrm{sample}}\;(\mathrm{mL})}

*V_sample = 1mL

Troubleshooting

Additional Resources

myLuminultra:
https://my.luminultra.com

Contact support:
https://my.luminultra.com/hc/en-us/requests/new