Design & Tech
Transforming Luminescence Assays: The Advanced Features of Luminescence 96
Illuminate your research with Luminescence 96, the world’s most compact luminescence plate reader. Designed with precision to enhance usability and flexibility, it is the ideal solution for luminescence-based assays in any laboratory setting. Its advanced features make luminescence research accessible, integrating 96 highly sensitive silicon photomultipliers and innovative technologies to deliver accurate, reproducible results.
Let’s dive deeper into the design features and technological innovations that make Luminescence 96 the ultimate tool for luminescence assays.
Reduced Luminescence Crosstalk for Your Assays
Luminescence assays are crucial to life science research, but crosstalk often compromises their sensitivity. Crosstalk occurs when luminescent signals from one well interfere with those in adjacent wells of a microplate, leading to inaccurate results.
Explore the intricacies of crosstalk and its implications for your luminescence assays.
The Luminescence 96 plate reader addresses this issue with its innovative design, including a compact footprint, a crosstalk reduction array, and an advanced shutter design. These features are meticulously engineered to minimize spatial crosstalk and prevent spectral overlap in luminescence assays, achieving crosstalk levels as low as 0.7 x 10⁻⁶. This ensures more precise and reliable results, making it an essential tool for high-sensitivity luminescence-based assays. Let's explore in detail.
Crosstalk reduction through compact design
Luminescence 96 leverages a compact footprint to enhance crosstalk reduction in luminescence assays. The innovative design allows precise microplate positioning within a narrow margin of just one-tenth of a millimeter from the housing of the detector units. This instrumental configuration is critical in minimizing device crosstalk, reducing the direct transmission of light between adjacent wells and the detectors.
Innovative array minimizes luminescence crosstalk
To further reduce crosstalk caused by the transmission of light between neighboring wells, Luminescence 96 incorporates a cutting-edge crosstalk reduction array (Fig. 1). Positioned on the device's interior optical surface, this array acts as a shield surrounding the microplate wells, effectively preventing light bleed from one well to another. This feature ensures precise detection and enhances the reliability of luminescence assay results.
Fig. 1: Illustration showing a crosstalk reduction array integrated into Luminescence 96 plate reader. This array prevents light from bleeding into neighboring wells, ensuring accurate luminescence readout from individual wells.
Pioneering shutter design and reading mechanism
With its unique shutter design, featuring a checkerboard pattern, Luminescence 96 selectively allows light to pass from specific wells while blocking any light leakage from adjacent wells. After completing a reading in particular wells, the shutter seamlessly transitions to the next position, facilitating the reading of neighboring wells while blocking the previously read ones (Fig. 2). This advanced reading mechanism ensures that luminescence crosstalk is effectively minimized.
Fig. 2: Luminescence 96 employs a unique checkerboard-patterned shutter design to selectively allow light from targeted wells while preventing leakage to adjacent wells. After each reading, the shutter moves to enable the next set of wells (position 2) while blocking the previously read ones (position 1), minimizing luminescence crosstalk.
By leveraging its distinctive features, Luminescence 96 achieves unparalleled low levels of crosstalk, setting a new standard for luminescence assay accuracy. To quantify the effectiveness of crosstalk mitigation, the Phosentix MicroQC™ plates were utilized. The MicroQC™ Plate is designed to evaluate device crosstalk, particularly in the lower right quadrant, where four luminescent wells with comparable intensity surround a central well (Fig. 3 A).
Crosstalk percentage was determined by measuring the signals from blank wells, central wells, and luminescent wells with Luminescence 96 using the formula: Crosstalk(%) = ((Center well – blank)/luminescence well)x100
Fig. 3: Illustration showing Phosentix MicroQCTM plate applied to quantify crosstalk and dynamic range of Luminescence 96 plate reader (A). For crosstalk evaluation, the lower right quadrant was utilized, which includes the designated center well (G11), luminescence wells (F11, G10, G12, H11), and blank wells (F10, F12, H10, H12). The measured luminescence crosstalk% of Luminescence 96 is presented, showing values well below the acceptance threshold criteria (B), highlighting its effectiveness in minimizing luminescence crosstalk.
According to the established MicroQC™ Plate crosstalk evaluation criteria, Luminescence 96 demonstrated outstanding crosstalk values below the stringent 0.02% threshold (Fig. 3 B). This exceptional performance underscores the device’s ability to minimize crosstalk, ensuring it can be effectively applied across a wide range of luminescence assays.
Eight Orders of Enhanced Dynamic Range
In luminescence assays, the dynamic range refers to the breadth of detectable light signal intensities. It defines the concentration window within a single experiment where quantification can be achieved.
With a linear dynamic range spanning up to 8 decades, Luminescence 96 emerges as a versatile tool for many luminescence assays. Integrated with highly sensitive silicon photomultipliers, Luminescence 96 offers flexibility through three distinct readout modes: photon counting, integration, and micro-integration.
- Photon counting: This mode focuses on individual photon detection events. Each detected photon generates a discrete electrical pulse, allowing for highly accurate counting of low-light signals.
- Integration: Unlike photon counting, in this mode, the electrical pulses are integrated across a defined time, allowing the detection of overlap pulses and increasing dynamic range. This approach enables Luminescence 96 to effectively measure different light intensities within the samples, ensuring versatility in luminescence analysis.
- Micro integration: Luminescence 96 features an integrated micro-sized pinhole within the shutter design to prevent saturation from higher light intensity signals. This pinhole restricts detection to just 1% of the signal when encountering high-light signals.
In Luminescence 96, all three modes are concurrently utilized to detect a wide range of signal intensities across the 96 wells. Detectors autonomously adjust parameters post-signal analysis, ensuring adaptive response based on the signal level in each well.
This approach facilitates user-friendly operation, eliminating the need for manual intervention in determining gain settings for measuring signals ranging from low photon counts to high-light intensities.
Fig. 5: The linearity of the Luminescence 96 Plate Reader was assessed using the Phosentix MicroQCTM plate. The readout was taken across the wells with varying signal intensities (row D, Fig. 3A), and the recorded luminescence values were compared to the reference values. The data demonstrated a strong correlation between the recorded and reference values, indicating that the Luminescence 96 Plate Reader exhibits excellent linearity and can reliably detect a broad range of luminescence intensities. This suggests that the Luminescence 96 has a high dynamic range, consistent with the performance recommended by the MicroQC plate.
The linearity and dynamic range of Luminescence 96 were rigorously validated using the Phosentix MicroQC™ plate. This specialized plate features various wells equipped with LEDs calibrated to standard reference levels, increasing intensities across the plate (Row D in Fig. 3 A). These LEDs simulate a range of light outputs typical for luminescence assays.
Luminescence signals from the MicroQC™ plate wells were measured using Luminescence 96 and compared against established reference values for quantitative assessment. A linear regression analysis yielded an R² value of 1.00, significantly exceeding the recommended threshold of 0.97, indicating a perfect correlation between the measured and reference values (Fig. 5). This finding demonstrates that Luminescence 96 possesses a high linear dynamic range and is capable of accurately measuring samples with varying levels of luminescent signal intensities.
Conclusion
Luminescence 96 stands at the forefront of luminescence plate reader technology, delivering unmatched crosstalk mitigation and dynamic range advancements. Meticulously designed for precision and efficiency, it addresses the critical challenges faced in luminescence assays, ensuring reliable and accurate data across various applications.
The device's advanced features—including a compact design, blocking array, and innovative shutter mechanisms—effectively reduce crosstalk to levels below 0.02%. Its high dynamic range enables accurate detection of luminescent signals while enhancing sensitivity, allowing for measuring a broad spectrum of light intensities.
In summary, Luminescence 96 represents a significant leap forward in luminescence technology, providing researchers with a robust, reliable, and user-friendly tool for their luminescence assays.
Are you interested in having more insights into Luminescence 96? Watch the unboxing video now!