White Microplates for Luminescence Enhanced Detection

Luminescence, a fascinating and versatile phenomenon, is widely used in both scientific research and diagnostics. It refers to the emission of light from a substance without the need for external heat or combustion. This unique characteristic makes luminescence an invaluable tool in various scientific fields, ranging from materials science and chemistry to biology and diagnostics.

Researchers often rely on luminescent signals to detect and measure various biological or chemical processes. Luminescent assays have become essential in studying cellular activities, detecting biomarkers, analyzing DNA sequences, and monitoring enzymatic reactions.

In these experiments and assays, the choice of detection platform is crucial for accurate and sensitive measurements. White microplates have emerged as an indispensable tool due to their ability to enhance luminescent signals effectively. The reflective properties of white microplates ensure that the emitted light is maximally captured by detection instruments, resulting in improved sensitivity and precision.

What is luminescence?

Luminescence occurs when a material absorbs energy and causes its atoms to become more energetic and excited. This absorption of energy can come from various sources such as light, electricity, or heat. When the atoms attain higher energy states, they become unstable and eventually need to return to their more stable and less energetic ground state. This transition back to the ground state is accompanied by the release of the excess energy in the form of light or other forms of radiation.

The Figure below illustrates the process of transitioning from one state to another through the absorption of energy, and subsequently returning to the initial state by releasing luminescent energy.

Depending on the type of energy different types of luminescence can occur:

Type	Definition
Chemiluminescence	Light is emitted as a result of chemical reactions
Bioluminescence	Light is emitted as a result of chemical reactions within living organisms (fireflies, glow-worms, and phytoplankton)
Electroluminescence	Light is emitted when an electrical current passes through a substance, such as a gas (LEDs, TV sets, and computer monitors)
Photoluminescence	Light is produced when light is shone on an object with luminous paint (through the absorption of photons, such as in watches that illuminate)
Röntgenoluminescence	Light is produced when an X-ray is shone on an object
Sonoluminescence	Light is emitted when high-energy sound waves pass through a liquid
Thermoluminescence	Light is produced when photons are emitted from a hot object
Crystalloluminescence	Light is produced during the process of crystallization
Triboluminescence	Light is emitted by pulling apart, ripping, scratching, rubbing, or altering crystals

 

Bioluminescence

Bioluminescence is a natural phenomenon that occurs in various organisms, ranging from marine creatures to fireflies. It refers to the ability of certain living organisms to produce and emit light.

One typical example of a bioluminescent reaction can be observed in the dinoflagellate known as Noctiluca scintillans, commonly referred to as “sea sparkle.”

When disturbed or agitated, Noctiluca scintillans exhibits a stunning display of bioluminescence. This single-celled organism contains specialized structures called scintillons, which contain luciferin molecules and an enzyme called luciferase. When these components come into contact with each other, a chemical reaction is triggered.

A typical example of a bioluminescent reaction is the reaction catalyzed by the firefly luciferase enzyme according to the following reaction:

The enzyme catalyzes the . Because ATP is required, firefly luciferases have been extensively used in biotechnology.

Chemiluminescence

A typical example of a chemiluminescent reaction is the reaction involving oxidized luminol in the presence of hydrogen peroxide in a basic environment.

In this reaction, luminol (C8H7N3O2) acts as the fluorophore, which is a molecule capable of emitting light when it undergoes certain chemical reactions. When luminol is oxidized by hydrogen peroxide (H2O2) in a basic environment, it transitions to an excited state.

This excited state is unstable and quickly returns to its ground state, releasing excess energy in the form of light. The emitted light appears as a blue glow or luminescence, giving rise to the term “chemiluminescence.”

This reaction can be represented by the following equation:

where:

• 3-APA (ground state) is the 3-aminophtalate
• 3-APA (excited intermediate state) is the fluorescing vibronic state of the 3-aminophtalate decaying to a lower energy level

A photomultiplier tube (PMTT) can be used to detect the light that is generated. This device converts photons into electrons. The resulting flow of electrons produces a current which directly correlates to the intensity of the light being detected. This current is measured in relative light units (RLU), providing a quantitative measure of the amount of light present.

Luminescence detection offers a simpler optical setup compared to fluorescence detection. Unlike fluorescence, it doesn’t require a dedicated light source or specific optics for excitation. This makes it an easier and more straightforward method for detecting luminescent signals.

Luminescence can exhibit different reactions, either as a “flash” or a “glow,” depending on the kinetic profiles involved. Flash luminescence produces an intense and brilliant signal, but it is relatively short-lived, typically lasting for only a few seconds. Glow luminescence is known for its stability, producing a consistent signal that can endure for extended periods of time, ranging from minutes to hours. On the other hand, flash luminescence tends to be less intense but requires a detection system with injectors. These injectors are designed to deliver a substrate to the reaction just before measurement, ensuring that the signal is not overlooked or missed. (see image below)

When it comes to luminescence assays, choosing the right type of microplate can significantly impact the accuracy and sensitivity of your results. White opaque microplates have emerged as a popular choice in this regard. These specialized plates are designed to reflect light, maximizing the signal and enhancing the overall performance of luminescence assays.

The reflective nature of white opaque microplates is particularly advantageous in situations where low-intensity signals need to be detected. When light is emitted from a sample within these plates, it encounters a reflective surface that redirects the photons back towards the detector. This redirection helps to capture more emitted light, resulting in increased signal strength and improved sensitivity.

In addition they minimize background noise by reducing unwanted stray light or interference from external sources. This ensures that the measured luminescent signal is more accurate and reliable.

Furthermore, white opaque microplates provide better heat resistance compared to transparent or clear plates. This quality is essential for maintaining temperature stability during prolonged assay incubations or when working with temperature-sensitive samples.

Biomat white microplates for biochemical immune luminescence assays

Biomat white microplates are a valuable tool in biochemical immune luminescence assays. These specialized microplates are designed to enhance the detection of luminescent signals, making them ideal for sensitive and accurate measurements in immunological research.

The unique white surface of Biomat microplates provides optimal reflectance and signal-to-noise ratio, allowing for the detection of even low levels of luminescence. This is particularly important in immune luminescence assays, where precise quantification of immune responses is crucial.

In addition to their exceptional performance, Biomat white microplates offer excellent compatibility with a wide range of reagents and detection systems commonly used in biochemical immune luminescence assays. This versatility allows researchers to choose the best combination of reagents and instruments for their specific experimental needs.

Furthermore, these microplates are manufactured using high-quality materials that ensure consistency and reliability across multiple experiments. They are designed to minimize well-to-well crosstalk and provide uniform distribution of samples throughout the plate, reducing experimental variability.

Biomat white microplates have become an indispensable tool for researchers in the field of immunology who require accurate and reproducible results. Their superior performance, compatibility, and reliability make them an excellent choice for biochemical immune luminescence assays.

Microplate Surface	Description
Medium-Binding	– These plates have a hydrophobic surface suitable for passive adsorption of proteins with different grades of hydrophobicity – Available in the 96-well breakable, strip and solid format
High-Binding	– These plates have a hydrophilic surface suitable for passive adsorption of proteins with different grades of hydrophilicity – Available in the 96-well breakable, strip and solid format
Carboxylated	– These plates are activated with carboxylic groups which can promote the covalent immobilization of biomolecules containing reactive free amino groups using EDC mediated amination – Available in the 96-well breakable, strip and solid format
Aminated	– These plates are activated with primary amino groups which can promote the covalent immobilization of biomolecules containing reactive groups such as carboxyl, thiol, or amino via well-known homo/hetero-bifunctional linkers, e.g. N-Hydroxysuccinimide (NHS) or Succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) – Available in the 96-well breakable, strip and solid format
Streptavidin	– These plates are activated with Streptavidin, a powerful and universal instrument for binding any biotinylated molecules (antibodies; antigens; proteins; peptides; polysaccharides; oligonucleotides; DNA fragments; etc.). They are used especially for molecules which do not offer reliable bonding by passive adsorption, or that adsorb in an unfavorable orientation. – Available in the 96-well breakable, strip and solid format
Streptavidin High-Binding	– These plates are activated with a dedicated form of Streptavidin that, besides having the same properties of basic Streptavidin, is particularly useful to set up competitive tests to measure biotinylated low molecular weight molecules – Available in the 96-well breakable, strip and solid format
A, G, and A/G proteins	– These plates are activated with Protein A, Protein G, or mixed A/G that provide alternatives to direct, passive adsorption methods for immobilizing antibodies for immunofluorescence plate-based assay techniques. They specifically bind to the Fc region of immunoglobulins of many mammalian species with an orientation that allows the F(ab)2 binding sites to be freely available for efficient binding to epitopes. – Available in the 96-well breakable, strip and solid format

Biomat white microplates for cell-based luminescence assays

Biomat white microplates have emerged as a valuable tool in the field of cell-based luminescence assays. These specialized microplates are designed to enhance the detection of luminescent signals emitted by cells, resulting in improved assay sensitivity and accuracy.

One of the key advantages of using Biomat white microplates is their ability to provide a high signal-to-noise ratio. This means that the luminescent signals generated by cells can be easily distinguished from background noise, allowing for more precise and reliable measurements.

The unique properties of Biomat white microplates also contribute to their effectiveness in cell-based luminescence assays. The white color of these plates helps to reflect light evenly across the entire well surface, ensuring uniform illumination and minimizing variations in signal intensity between wells.

Furthermore, Biomat white microplates are compatible with a wide range of luminometers and plate readers, making them versatile tools for researchers working with different detection systems.

In summary, Biomat white microplates offer significant advantages for cell-based luminescence assays. Their ability to enhance signal detection and provide a high signal-to-noise ratio makes them an ideal choice for researchers seeking improved assay sensitivity and reliable results.

Microplate Surface	Description
Tissue Culture Treated (polystyrene)	– TC-treated plates guarantee that the surface chemistry offers a uniform surface with both hydrophilic and negative-charge properties; this treatment will lead to increased cell attachment – The bottom can be either opaque or transparent – Sterilized with lid – Available in 96- and 384-well formats
Poly-D-lysine and Poly-L-lysine	– With some cell types, to enhance cell attachment, growth, and differentiation, the plastic support must have a thin layer of polymer polycationic that helps the cells, or tissues, to adhere to the surface. This enhanced surface is obtained by coating it with Poly-lysine, a synthetic, positively charged polymer. – The bottom can be either opaque or transparent – Sterilized with lid – Available in 96- and 384-well formats