Fluorescence Applications

Discover the power of Fluorescence. From life sciences to materials, including water and forensics, Fluorescence is found in many fields of applications.

Protein Quantification at Low Concentration using Fluorescence and Absorbance
Fluorescence Emission spectra of transferrin at increasing concentrations
In this application note, we highlight the high sensitivity of the Duetta for measuring transferrin at very low concentrations. The Duetta spectrofluorometer combines both fluorescence and absorbance measurements simultaneously, thus providing key advantages for protein analysis. In parallel, we compare the fluorescence results obtained using the Duetta with absorbance results obtained with a dedicated low-volume, benchtop spectrophotometer.
The Power of Duetta and EzSpec Software for Biological Applications
Protein A280 application
Duetta is an analytical instrument capable of recording both the fluorescence and the absorbance of a sample. Absorbance data provides concentration-dependant information on non-fluorescent compounds. Through its user-friendly software, Duetta allows different measurements related to biomolecules like DNA, RNA and proteins including protein concentration, DNA/RNA purity, finding the concentration of an unknown sample based on a standard-based calibration curve, Pass/Fail testing and all standard fluorescence and absorbance related measurements. All these functionalities are possible through dedicated applications in EzSpec™ software for Duetta.
Harmful Algal Bloom Species
This study describes the application of simultaneous absorbance and fluorescence excitation-emission matrix (EEM) analysis for the purpose of identification and classification of freshwater planktonic algal species.
A-TEEM Fluorescence for Identification and Quantification of Fulvic Acid Adulteration in Commercial Humic Products
This app note demonstrates that the A-TEEM molecular fingerprinting technique can easily distinguish between different components in mixtures that contain FF and other ingredients considered to be adulterants, including lignosulphonates, organic acids, molasses, and provides a quantitative evaluation of mixtures.
Fluorescence Spectroscopy and Water Quality
Water travels through the environment carrying dissolved organic matter (DOM), made up of various chemical compounds, which have entered the water column from many sources.
Sulfur monitoring in Marine Oil with the SLFA in a growing environmental protection context
AN45 - Sulfur monitoring in Marine Oil with the SLFA in a growing environmental protection context Illustration
Monitoring of Sulfur in petroleum products is an important topic in the current environmental protection context. Indeed, during combustion, Sulfur will cause acid rains and particles emissions that induce environmental and health problems. If industries and vehicles emissions have been the main target of Sulfur emission control since few tens of year, the new target is the maritime navigation. In this domain, now, the level of Sulfur is regulated by the MARPOL (International convention) Annex VI. For this specific regulation, the technique and measuring range of our SLFA series is perfectly suitable to allow the fast and accurate determination of Sulfur in Marine oil. Our compact SLFA-60 can be used on board and the SLFA-6800, with its autosampler (eight positions), will enhance the productivity of the control laboratories.
Studying heart disease by simultaneous measurement of changes of Sarcomere Length
Studying heart disease by simultaneous measurement of changes of Sarcomere Length
Studying heart disease by simultaneous measurement of changes of Sarcomere Length and concomitant intracellular ionic calcium concentration in freshly isolated cardiac myocytes.
Time‐resolved Fluorescence for Monitoring Food Composition
The use of time‐resolved fluorescence has expanded as the relative cost of instrumentation has decreased in recent years. One area where this is especially true is in the food industry, where time‐resolved fluorescence has been applied in the characterization of food stuffs as well as aspects related to food safety and degradation.
Liquid in situ Fluorescence Measurements
No need to sample—simply take your readings directly in place in locations such as reaction vessels Collecting samples during a continuous process can be tedious and even dangerous if hazardous materials are involved.
PLQY in NIR with Fluorolog-QM and K-Sphere
The Fluorolog-QM fluorescence spectrometer with an integrating sphere option is an excellent choice for PLQY measurements in NIR. The K-Sphere is very convenient and easy to use, as it couples directly to the sample compartment optics and allows a use of external light sources, such as DPSS lasers, which can be attached to the front of the sample compartment. The sphere has holders for cuvettes, slides and powders that can be easily interchanged. The results demonstrate very good reproducibility and precision for NIR PLQY measurements spanning almost two orders of magnitude. Based on multiple measurements, the demonstrated PLQY standard deviations range from 1.3% for high QY value (77%) to 6.4% for the QY below 1%.
Milk compounds characterization by optical spectroscopies and laser diffraction
In the food industry, the compounds characterization is a critical step to ensure the quality of the products or to provide information to customers which can be sensitive to allergies. In this application note, we showed how optical spectroscopies and laser diffraction can help for food compounds characterization, especially on a specific product, i.e. milks.
Photoluminescence Upconversion with the Fluorolog-QM
DPSS laser mounted to the front of the Fluorolog-QM sample compartment
The new Fluorolog-QM spectrofluorometers, due to their modularity and advanced software and a universal interface, are an ideal choice for studying multiple aspects of upconversion. This technical note illustrates the use of Fluorolog-QM-75-21 for spectral and time-resolved characterization of these materials.
Insulin Structure and Stability Assessment
Multivariate analysis of different insulin proteins
Stability and aggregation of insulin are studied using simultaneous fluorescence excitation emission matrices (EEMs) and UV-Vis absorbance spectroscopy [1]. Insulin is a protein-hormone, produced by the pancreas and is necessary for basic metabolic processes. The different types of commercial insulin therapeutics generally fall into two categories: short-acting and long-acting insulin. The difference between some short-acting and long-acting insulin is, in some cases, only one to three residues in the protein sequence.
Classification and Phenolics Analysis of Red Wines with A-TEEM Molecular Fingerprinting
Classification and Phenolics Analysis of Red Wines with A-TEEM Molecular Fingerprinting
Aqualog®, an analytical instrument based on the simultaneous measurement of Absorbance, Transmission, Fluorescence Excitation and Emission Matrix (A-TEEM). A-TEEM fingerprints yield qualitative and quantitative composition of key flavor and color determinants in wine and spirits that are not discernible with simple Absorbance or Transmission data analysis.
Protein A280 for Protein Concentration
The new HORIBA Duetta fluorescence and absorbance spectrometer offers many unique benefits for molecular spectroscopy. It is primarily thought of as a spectrofluorometer that combines absorbance and fluorescence spectroscopy to correct the fluorescence fingerprint for concentration-related effects. Duetta can also be used as a precise absorbance spectrometer. This application note explores the use of Duetta for a common absorbance application, the Protein A280 application.
Rapid Extra Virgin Olive Oil Classification and Blend Quantitation
Rapid Extra Virgin Olive Oil Classification and Blend Quantitation
The resurgence of interest in the Mediterranean Diet and its associated health benefits have directed focus on the role that Extra Virgin Olive Oil plays. The increasing awareness is leading to increasing product demand, but also opportunities to compromise quality. Hence the need for rapid analytical methods to perform Quality Analysis of various product samples.
Fluorolog-QM Enhancement with a Broadly-Tunable OPO Pulsed Laser
The Fluorolog-QM equipped with the optional OPO-based Opolette laser provides the ultimate tool for time-resolved measurements in the microsecond to millisecond time domain for spectral ranges spanning from UV to NIR.
Elucidating Local Viscosity Using Fluorescence Lifetime Measurements
Certain fluorescent molecules, known as molecular rotors, can be employed to estimate the local (nanoscale) viscosity in microheterogeneous systems by measurement of their fluorescence lifetime. This can be advantageous over the usual fluorescence anisotropy method, as the measurement is simpler and faster to perform. This is demonstrated using the HORIBA Scientific TemPro fluorescence lifetime system to monitor the gelation of silica produced using the sol‐gel technique.
How Inner-Filter Effects (IFE) can affect your fluorescence measurements: A case study - Colloidal InP Quantum Dots
During the last two decades, a great deal of attention has been focused on the optoelectronic properties of nanostructured semiconductors or Quantum Dots (QDs).
Detection of Explosives with Fluorescence
International terrorism, security concerns, and the remains of forgotten landmines throughout the world have increased interest in detection of explosive materials.
Studying perovskite solar cells with HORIBA Scientific equipment
With their ~20 % efficiency, hybrid perovskite solar cells are the new promising candidate for next generation photovoltaics. Thanks to the wide HORIBA Scientific portfolio, different techniques can be used to gain in depth knowledge on the optoelectronic properties and mechanisms of this class of materials. In this application note we decided to use spectroscopic ellipsometry, steady-state and time-resolved fluorescence and Glow Discharge Optical Emission Spectroscopy to investigate the properties of CH3NH3PbI3 thin films deposited on a spin-coated PEDOT:PSS. The impact of the exposure to air was addressed.
Characterizing Tooth Decay with Fluorescence
Quantum Dot Absorbance, Photoluminescence Spectra and Lifetimes
A-TEEM™ for Qtracker® 655 quantum dots
Quantum dots (QDs) are semiconducting spheres in the size typically in the range of 1 to 10nm. The size of these small spheres give quantum dots the semiconducting properties and resulting photoluminescence that would not necessarily occur for the same material on larger scales.
The Measurement of Singlet Oxygen Lifetime Sensitized using Rose Bengal
The study of singlet oxygen (1O2) is of interest, principally, as it is a highly reactive species. It can be produced by photosensitisation, usually of a molecule such as a dye or porphyrin. Thus, by the appropriate selection of sensitiser, the presence of oxygen and light, 1O2 can be selectively generated. From a biological aspect it has the ability to damage and destroy cells, which has lead to interest in its use as an anticancer agent in photodynamic therapy (PDT).
Spectroscopic Analysis of Red Wines with Aqualog
Spectroscopic Analysis of Red Wines with Aqualog
HORIBA’s Aqualog is uniquely equipped with patented simultaneous Absorbance spectral and fluorescence excitation-emission matrix (EEM) technology which provides rapid access to a wide range of parameters important to commercial wine processing and quality characterization.
The NanoLog Series: A New Generation of Performance
The NanoLog has a reputation as the premier instrument for the exploration of single-walled carbon nanotubes (SWCNTs).
Recording Fluorescence Quantum Yields
When a fluorophore absorbs a photon of light, an energetically excited state is formed. The fate of this species is varied, depending upon the exact nature of the fluorophore and its surroundings, but the end result is deactivation (loss of energy) and return to the ground state.
Measuring Silica Nanoparticles via Fluorescence Anisotropy
Silica is currently one of the most important industrial materials, whose nanoparticles are formed via a sol-gel process.
Measuring PL Upconversion Spectra and Lifetimes of Lanthanide-Doped Nanoparticles
Upconverting lanthanide-based nanomaterials exhibit a unique fluorescence anti-Stokes shift, which enables them to convert NIR wavelength excitation into visible shorter wavelength emissions (NIR to UV-Vis).
Plasmon enhancement of protein fluorescence by silver nanostructures
The use of metal surfaces in conjunction with fluorescence molecules employing a plasmon effect, sometimes referred to as metal enhanced fluorescence, can be advantageous because of the possible enhancement of photophysical properties.
Nanophotonics with Fluorescence Instruments
HORIBA Jobin Yvon’s spectrofluorometers have many applications in nanophotonics research: single-walled carbon nanotubes (SWNTs), quantum dots (QDs), and organic light-emitting diodes (OLEDs). Quantum confinement affects nanomaterials’ photoluminescence: when the semiconducting nanoparticle is smaller than the bulk material’s Bohrexciton radius, the bandgap energy is inversely proportional to the nanoparticle size.
Time‐resolved luminescence of security inks from the UV to NIR
The use of security features, such as luminescent inks, has increased significantly in an attempt to prevent fraud and counterfeiting of materials and goods.
Fluorescent Pigments in Living Coral
The brightly-colored coral reefs that make scuba-diving and snorkeling so enjoyable are essential to the survival of much underwater life. Not only do reefs offer a haven for smaller fish to hide from larger predators, but also some fish actually survive by eating the reefs themselves. Reefs offer protection to plants and animals from the ravages of waves and ocean currents. Thus, when the reefs die, so do many other living creatures.
Assessing UV Damage of Hair with Fluorescence
Fluorescence from hair
The results of UV exposure of human hair can be quantified with fluorescence spectroscopy. This is of significant interest to cosmetics and pharmaceutical companies that manufacture hair creams, coloring, shampoos, and conditioners.
Better Data on Carbon Nanotubes with the NanoLog
Improvements to the HORIBA Scientific NanoLog®, already the best spectrofluorometer for exploration of single-walled carbon nanotubes (SWCNTs), render it even more suitable for this application.
Monitoring Whole Leaf Fluorescence Using Time‐resolved Techniques
Light incident on a leaf can be absorbed by chlorophyll to commence the photosynthetic cycle. Excess energy can be liberated as heat or by emission of fluorescence and this can be used to assess the efficiency of the photosynthetic process.
Spectroscopic Methods for Sunscreens Characterization
This Application Note outlines three different kinds of spectroscopic tools being used for the characterization of sunscreens, and discusses the obtained results. These include Fluorescence spectroscopy for photoactivity, Particle Size analysis for composition and Raman microscopy for formulation investigation.
Monitoring Cell Culture Media Variability using a Simple Optical Technique (A-TEEM Molecular Fingerprinting)
Monitoring Cell Culture Media Variability using A-TEEM Spectroscopy
With the rise of protein production using mammalian cell culture, it has become increasingly important to control the quality of the cell culture media for use in production processes. Cell culture media are usually prepared as aqueous solutions and should provide everything a cell line needs for optimal cell growth as well as product yield and quality.
Effect of temperature on HSA structure inferred using timeresolved room-temperature phosphorescence
Selection of SpectraLED pulsed excitation sources
To access intrinsic amino acids, such as tryptophan, as probes, the UV excitation wavelengths for pulsed phosphorescence measurements have long been the preserve of low-repetition-rate gas-filled lamps or larger laser systems. Recent developments have enabled the use of interchangeable semiconductor diodes...
Measurement of carrier lifetime in perovskite for solar cell applications
Hybrid perovskite photovoltaics (PV) show promise because of their good efficiencies, which can be around 20%. Along with their PV characteristics, perovskite materials exhibit a high degree of radiative recombination.
Photoluminescence Lifetimes in NIR
Applications that involve photoluminescence (PL) measurements in NIR have been rapidly growing in recent years. The demand comes mainly from several areas in materials science, such as fiber optics telecommunication, solar energy conversion, lasing media, LED and OLED technologies, and development of upconversion nanoparticles for biomedical analyses and bioimaging.
Photoluminescence Spectroscopy of Quantum Dots
Photoluminescence Spectroscopy of Quantum Dots
Quantum dots (QDs) have potential applications in optoelectronics, biosensing, biolabeling, memory devices, and sources of laser light.
Flat Panel Displays and Fluorescence
One of the fastest-growing segments of the semiconductor industry is concerned with a new generation of graphic displays for communications and high-definition television sets. For phosphors that might be used as the active medium in such displays, the critical characteristics are the lifetimes and wavelengths of their emissions.
Near-IR Photoluminescence of Quantum Dots
HORIBA Jobin Yvon’s NanoLog® spectrofluorometer, specially optimized for recording near-IR fluorescence from nanoparticles, includes a double-grating excitation monochromator, imaging emission spectrograph with a selectable-grating turret, and a variety of detectors.
MCS and Protein Phosphorescence
Multichannel scaling (MCS) single-photon-counting spectroscopy performed using HORIBA Jobin Yvon’s FluoroCube fluorescence lifetime system.
Tryptophan phosphorescence within protein molecules is gaining attention as a probe of protein dynamics and structure. The tryptophan phosphorescence lifetime, τ, varies with the protein molecule’s local environment and conformation.
Upconversion of Lanthanide-containing glasses using DD‐980L excitation
The phenomenon of upconversion is an optical process that takes in lower energy (longer wavelength) photons and emits higher energy (shorter wavelength) photons.
Better Signal-to-Noise Ratios for Carbon Nanotube Spectra
Better Signal-to-Noise Ratios for Carbon Nanotube Spectra
Corrected emission spectra1 of carbon nanoparticles can provide excitation–emission matrices (EEMs) for a range of excitation wavelengths.
EPA Stage 2 Disinfection Compliance with Aqualog
This application note describes the use of the Aqualog for monitoring regulated Dissolved Organic Matter (DOM) and disinfection by-product issues for drinking water treatment.
Characterizing Lanthanides in Glasses for Optical Applications
Glasses are essential materials with a multitude of uses and many forms. In the area of optoelectronics there is an interest to modify the glass composition to favor the incorporation of lanthanide elements.
Fluorescence Spectra from Carbon Nanotubes with the NanoLog
Fluorescence Spectra from Carbon Nanotubes
Single-wall carbon nanotubes (SWNTs), consisting of rolled-up single sheets of carbon atoms, have received much attention recently.
Characterizing Galvanizing Bath with Fluorescence
Automobile-parts production can be paralyzed by a defective electrongalvanizing (EV) bath line.
Visualizing local viscosity using fluorescence lifetime microscopy
DynaMyc with FLIM (fluorescence lifetime imaging)
The use of fluorescent molecules, known as molecular rotors, is advantageous in estimating the local (nanoscale) viscosity in microheterogeneous systems, since it just requires the measurement of their fluorescence lifetime.
Fast & Non-invasive Determination of Skin State
Fast and non-invasive determination of skin
Biological tissues contain chromophores that absorb light, as well as fluorophores that absorb and reemit light (fluorescence effect). Light absorption depends on the chromophores’ content and their distribution within the organic matter.
Visualizing dental caries using fluorescence lifetime microscopy
Teeth are naturally fluorescent and changes in their composition, caused by decay for example, affect their fluorescence behavior.
Endogenous Skin Fluorescence In Vivo on Human Skin
The fluorescence spectra of intrinsic protein fluorophores have been studied extensively and used in investigating biological events.
Fluorescence on Small or Solid Samples
Samples can be valuable. Either you don’t want to waste them, or you may not have very much starting material. Biological proteins and enzymes, for example, are often obtained in small volumes and may be expensive.
Detecting Conformational Rotamers via TCSPC
Detecting Conformational Rotamers via TCSPC
Among the possible fluorescence biosensors for medical and biochemical monitoring and imaging are the flavonoids, compounds that occur in many plants and their products, such as tea, chocolate, and red wine.
Kinetic Fluorescence Determination of Vitamin B1
This technical note describes kinetic fluorescence as an analytical technique to quantify non-fluorescent species. The technique is applied to the determination of thiamine (vitamin B1) in solution.
FRET with a HORIBA Phosphorimeter
Luminescence spectra of peptide + terbium
This Technical Note describes an example of Förster resonance energy transfer (FRET) from a peptide-terbiumcomplex donor to a fluorescein acceptor, using the HORIBA Jobin Yvon phosphorimeter.
Investigating photocleavage using time‐resolved emission spectra
The choice of protecting group is of crucial importance in the success of many steps in organic synthesis and the manipulation of polyfunctional molecules, since they can prevent the formation of undesired side products and reactions.
Noninvasive In-Vivo Determination of Sunscreen-UVA Protection Factors
Noninvasive In-Vivo Determination of Sunscreen-UVA Protection Factors
The development and evaluation of UVA (320–400 nm) sunscreens is important because UVA sunlight can penetrate deep into human skin and cause severe internal damage, as well as erythema and photoaging.
Holistic Analysis of Mammalian Cell Proliferation using Fluorescence Spectroscopy
Holistic Analysis of Mammalian Cell Proliferation using Fluorescence Spectroscopy
Stopped flow time‐resolved fluorescence study of serum albumin – curcuminoid binding
Rapid mixing accessories to perform stopped flow measurements have found application in characterizing interactions and reactions occurring in solution. Reactants are expelled from syringes, mixed and injected into a flowcell.
Plasmon enhancement of protein fluorescence by silver nanostructures
The use of metal surfaces in conjunction with fluorescence molecules employing a plasmon effect, sometimes referred to as metal enhanced fluorescence, can be advantageous because of the possible enhancement of photophysical properties. For example, the emission intensity of the fluorophore, can be improved.
Dye‐protein binding monitored in a microliter volume using timeresolved fluorescence
The potential health benefits stemming from the antioxidant activity of curcumin, commonly found in turmeric (Curcuma longa L), has attracted the interest of several research groups.
Selective excitation of tryptophan fluorescence decay in proteins using a subnanosecond 295 nm light-emitting diode and time-correlated single-photon counting
We demonstrate an AlGaN light-emitting diode (LED) giving pulses of ~600 ps full width half maximum, 0.35 µW average power, 0.6 mW peak power, and ~12 nm bandwidth at 295 nm. This source is ideal for protein intrinsic tryptophan fluorescence decay research without the unwanted excitation of tyrosine and paves the way to lab-on-a-chip protein assays using fluorescence decay times. Fluorescence decay and anisotropy decay measurements of human serum albumin are reported and the usefulness of the 295 nm LED demonstrated in comparisons with a nanosecond flashlamp and LEDs with nominal wavelength emission of 280 nm.
Fluorescence Anisotropy Studies
Polarized light striking a fluorescent molecule results in polarized fluorescence. This polarized emission gradually returns to unpolarized fluorescence depending on rotational diffusion and other factors. Anisotropy is directly related to the polarization, and is the ratio of the polarized light component to the total light intensity.


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