3D Cell-ATP Viability Detection Kit 

MCE 3D Cell-ATP Viability Detection kit is based on high-sensitivity bioluminescence detection technology (luciferase system) that quantitatively measures ATP to detect the number and viability of 3D cells in culture. By directly adding the reagent to the cell culture, the cell aggregates are lysed, releasing ATP and generating a stable “glow-type” luminescent signal. The intensity of the luminescent signal is proportional to the ATP content within a certain range, indirectly reflecting the number of viable cells in the sample, and can be used for the quantitative detection of viable cell counts.

Features of MCE 3D Cell-ATP Viability Detection kit

Ready-to-Use: No need to digest or lyse cells and organoids. The detection reagent has been optimized for enhanced lysis capability, effectively penetrating the interior of the spherical structures formed in 3D cultures.

User-friendly: No washing, replacing, or removing of culture medium is required before detection. Simply add the detection reagent and incubate at room temperature for 30 min before proceeding with the assay.

Stability: The “glow-type” luminescent signal is more stable, with the luminescent signal lasting up to 3 h.

Flexible detection scheme: Suitable for various types of plate assays.

Wide applicability: Suitable for detecting small sample volumes and high-throughput screening of large sample sets.

-20°C, 1 year

Keep away from light and avoid repetitive freeze-thaw cycles.

1. This product contains luciferase, which is sensitive to temperature. Repeated freeze-thaw cycles may lead to gradual loss of activity. It is recommended to aliquot and store the product after the first thaw to avoid repeated freeze-thaw cycles. Ensure that the aliquot containers are free from ATP contamination.

2. After thawing, the product can be stored at 4°C, protected from light, and remains stable for 2 days.

3. Luciferase is sensitive to temperature. Before the assay, both cells and detection reagents should be equilibrated to room temperature.

4. High solvent content in the test drug may interfere with the luciferase reaction, affecting the chemiluminescence signal. To eliminate solvent interference, it is recommended to set up control wells containing the solvent in the cell culture medium.

5. The best detection results can only be obtained when the cell aggregates are fully mixed with the viability detection reagent and completely lysed. If the cell aggregates are not fully lysed, resulting in uneven luminescence values between wells, optimize the procedure by increasing the shaking amplitude or extending the incubation time.

6. For detection, use a white or black 96-well or 384-well plate. Using regular transparent 96-well or 384-well plate may cause interference between adjacent wells. Additionally, ultra-low binding culture dishes or multi-well plates specifically designed for 3D cultures can be used to culture the 3D cells and then transferred to regular white or black multi-well plates for detection.

7. This product is for R&D use only, not for drug, household, or other uses.

8. For your safety and health, please wear a lab coat and disposable gloves to operate.

For a 96-well plate (with 100 μL of cell culture medium per well), add 100 μL of the cell viability detection reagent, maintaining a ratio of V_{culture medium} : V_reagent = 1:1. After thawing the product, take an appropriate of the reagent and equilibrate it to room temperature before use.

Note: It is recommended to aliquot the product after the first thaw to avoid repeated freeze-thaw cycles.

Treat the cells with the drug according to the experimental conditions, and culture the 3D cells under the appropriate conditions.

1. Take the cell culture plate to be tested and equilibrate it at room temperature for 30 min.

2. Add 100 μL of the viability detection reagent to each well, gently shake at room temperature for 5 min to promote complete cell lysis.

3. Incubate the cell culture plate at room temperature for 25 min to allow the luminescent signal to stabilize.

4. Use a multifunctional microplate reader with chemiluminescence detection capability to measure the chemiluminescent signal. Record the luminescence value, with a typical detection time for each well is 0.25 - 1 s.

Note: a. Specific parameters can be adjusted based on the type of instrument and the detection sensitivity.

b. The penetration of the viability reagent into 3D cells and the time required for the luminescent signal to stabilize are related to the cell type, culture time, and the size of the 3D cell aggregates. The shaking step can be replaced with repeated pipetting.

c. Detection can begin directly after adding the viability reagent → shaking for 5 min → incubation for 5 min. Measure and record the luminescence value every 5 min until the luminescent signal stabilizes, and use the luminescence value at the stable point as experimental data.

1. This product contains luciferase, which is sensitive to temperature. Repeated freeze-thaw cycles may lead to gradual loss of activity. It is recommended to aliquot and store the product after the first thaw to avoid repeated freeze-thaw cycles. Ensure that the aliquot containers are free from ATP contamination.

2. After thawing, the product can be stored at 4°C, protected from light, and remains stable for 2 days.

3. Luciferase is sensitive to temperature. Before the assay, both cells and detection reagents should be equilibrated to room temperature.

4. High solvent content in the test drug may interfere with the luciferase reaction, affecting the chemiluminescence signal. To eliminate solvent interference, it is recommended to set up control wells containing the solvent in the cell culture medium.

5. The best detection results can only be obtained when the cell aggregates are fully mixed with the viability detection reagent and completely lysed. If the cell aggregates are not fully lysed, resulting in uneven luminescence values between wells, optimize the procedure by increasing the shaking amplitude or extending the incubation time.

6. For detection, use a white or black 96-well or 384-well plate. Using regular transparent 96-well or 384-well plate may cause interference between adjacent wells. Additionally, ultra-low binding culture dishes or multi-well plates specifically designed for 3D cultures can be used to culture the 3D cells and then transferred to regular white or black multi-well plates for detection.

7. This product is for R&D use only, not for drug, household, or other uses.

8. For your safety and health, please wear a lab coat and disposable gloves to operate.