FluoroMax® Plus: 5th Generation

World-renowned FluoroMax Benchtop Spectrofluorometer with 30 years of innovation and improvement.

FluoroMax Plus Series: 30 Years of Innovation and Improvement

The FluoroMax® Plus Series represents the culmination of decades of HORIBA's industry-leading experience in developing high-performance spectrofluorometers in a convenient, affordable, and easy-to-use benchtop model. Featuring exquisite, optically perfect, all-reflective optics combined with photon counting detection and a multitude of accessories, the FluoroMax Plus offers superior sensitivity and versatility for any tabletop spectrofluorometer.

The FluoroMax Plus can be enhanced to suit a broad range of luminescence experiments with a variety of optional accessories to expand capabilities and performance, including:

• An affordable LN-cooled NIR InGaAs detector for spectral detection up to 1,700 nm.
• Enhanced TCSPC fluorescence lifetime detectors for very short (down to 25 ps) lifetime measurements.
• Extended NIR PMTs for spectral and TCSPC detection up to 1,700 nm.
• PLQY integrating spheres.
• Phosphorescence pulsed light source and detection.
• Automated microwell plate reader for high sample throughput.
• Fluorescence microscopy, including PL mapping.

The FluoroMax Plus is ideal for measuring solid and liquid samples, supporting high throughput screening, cryogenic or elevated temperatures, absolute quantum yields, microliter volumes, stopped flow mixing or titration, and micron scale measurements using microscopes. With an extensive list of accessories, the FluoroMax Plus offers unparalleled flexibility to meet diverse experimental needs.

Future Ready: All accessories can be added to your system at any time as your needs change or funds become available.

• Research Level Performance
• Enhanced Modularity
• Perfect for any Application!

With thousands of units operating in universities and research labs globally, the FluoroMax Plus is considered the reference benchtop spectrofluorometer. The FluoroMax Series has proven itself ideal for demanding steady state, time-resolved, TCSPC, PLQY, and NIR luminescence needs.

The FluoroMax Plus is manufactured at HORIBA's state-of-the-art, 132,000 square foot facility in Piscataway, New Jersey, ensuring the highest quality manufacturing processes and efficiencies. Trust the FluoroMax Plus for your most demanding steady state, time-resolved, TCSPC, PLQY, and NIR luminescence requirements.

Made in the USA ??

Performance by Design

Maximizing Signal, Minimizing Noise

Comparing spectrofluorometer sensitivity can be confusing due to multiple specifications and definitions. The FluoroMax Plus offers the highest sensitivity available in a benchtop model. Sensitivity is often reported by minimum measurable concentration or the signal-to-noise ratio (S/N) of the water Raman scattering peak. HORIBA uses the First Standard Deviation (FSD) method for reporting water Raman S/N, which is applicable to photon counting fluorometers.

The FluoroMax Plus is a photon counting spectrofluorometer, offering superior sensitivity compared to analog-based PMT fluorometers. The FSD signal-to-noise ratio formula is provided, based on measuring the peak signal at the water Raman peak (397 nm for 350 nm excitation) and the noise in a region without Raman emission (450 nm). Acquisition parameters for water Raman measurements include:

• Excitation: 350 nm with 5 nm bandpass
• Emission scan: 365–455 nm with 5 nm bandpass
• Emission wavelength step interval: 0.5 nm
• Integration time per step: 1 second
• Single emission scan (no repeats)
• No smoothing of data points
• No optical filters

A description of a Raman band measurement of ultrapure water in a 1 cm quartz cuvette, qualified using the FSD method with a standard Xenon lamp source, is provided. Another description details Coumarin 153 diluted to 5.0 pM in Ethanol, with the spectrum showing subtraction of blank data to remove solvent background and Raman scatter.

HORIBA Quality Optics

The FluoroMax Plus achieves its high sensitivity through an optimized design featuring all-reflective optics, photon counting detection, and reduced stray light via HORIBA Jobin Yvon quality gratings and optical elements.

Measure Smaller Samples, Detect Smaller Changes

Big Discoveries Come from Small Changes

Utilizing the most sensitive fluorometer provides confidence in detecting small changes in data, crucial for driving research and discovery. This sensitivity allows for measurement of more dilute samples or requires less sample volume, saving time and money on sample preparation.

Sensitivity Means Speed

The true limit on data acquisition speed is not the monochromator grating scan speed, but the time needed at each wavelength for an acceptable signal-to-noise ratio. The FluoroMax Plus's higher sensitivity enables faster measurements, especially for weaker samples.

Dynamic Range

The FluoroMax Plus handles samples with varying fluorescence intensities, measuring strong and weak peaks within the same scan. Its intra-spectral range of over 6 orders of magnitude simplifies kinetics studies by eliminating the need to repeat measurements at different integration times.

A spectrum of a mixture of terbium chloride and europium chloride in ultrapure water is described, with an inset showing a zoomed view of the weak europium chloride peak measured alongside the strong terbium chloride peaks. Spectra of pure terbium chloride and europium chloride are also shown for reference.

Enhanced Measurements

Absolute Quantum Yield Measurements

Quantum yield is a key parameter in photoluminescence, critical for applications like new material development and photovoltaics. It is defined as the ratio of emitted photons to absorbed photons. HORIBA is recognized for its superior performance in PLQY measurements.

The QuantaPhi-2 is an accessory for internal photoluminescence quantum yield (PLQY) and CIE measurements. It features an all-reflective sphere for sample placement, enabling direct quantum yield measurement of solids, powders, or solutions by comparing sample and blank measurements. The QuantaPhi-2 includes a 121 mm diameter Spectralon® integrating sphere with excellent reflectivity from 250 to 2500 nm, and accommodates powders, solids, and solutions.

Specialty integrating spheres are also available. High-precision absolute quantum yield characterization of a quantum dot sample is described, enabled by the QuantaPhi-2 integrating sphere. FluoroMax software also automatically generates CIE 1931 and 1976 colorimetric values.

Kinetics

Measuring temporal variations in fluorescence spectra provides insights beyond single time point measurements, such as monitoring protein stability and folding. The FluoroMax Plus Series allows dynamics measurements from milliseconds to hours.

A description of kinetics of lysozyme dissolved in guanidinium hydrochloride, monitored by tryptophan fluorescence over 1 hour, with data corrected for photobleaching, is provided.

Absorbance/Transmittance Accessory

Add absorbance capabilities to fluorescence measurements. Featuring IFE Correction, the FluoroMax Plus can correct Excitation Emission Matrices (EEMs) for the inner filter effect (IFE), providing a true molecular fingerprint. IFE correction is essential for accurate chemometric analysis.

A description of EEMs of gallic acid at different concentrations, with and without absorbance corrections (A-TEEM™), is provided. The corrected spectra show improved linearity at higher concentrations compared to traditional fluorometers.

Polarization

Optional polarizers allow for the study of molecular rotation, providing information on sample aggregation, structural changes, and molecular binding. The FluoroMax Plus uses Glan-Thompson prisms for high efficiency across the UV to NIR range.

A description of fluorescein dissolved in sucrose solutions is provided, showing how increased temperature (and decreased viscosity) yields faster rotation times and lower anisotropies. Anisotropy is useful for understanding macromolecule shape and binding.

Technologies and Applications

The FluoroMax Plus covers the broadest range of luminescence research. A visual representation lists various technologies and applications in a hexagonal format:

• Fast Mixing Reaction
• Polarization and Anisotropy
• Ratiometry
• Quantum Yield
• Plate Reader
• Temperature Ramp
• 3D EEM Matrix
• Phosphorescence
• Absorption and Transmission
• Up Conversion
• Fluorescence
• Titration
• Time-based Kinetics
• Fluorescence and Phosphorescence Lifetimes
• Microscopy
• Cryogenics

Application areas are also depicted in a hexagonal format, including:

• Nanomaterials
• Chemistry
• Environment
• Pharma
• Analytical Sciences
• Photovoltaics
• Petrochemicals
• Material Research
• Medical and Bioscience
• Polymers
• Geology
• Food Science
• Cosmetics
• Forensics
• Rare Earths

TCSPC: Take Your Measurements to the Next Dimension with Fluorescence Lifetimes

Time Correlated Single Photon Counting

Add lifetime measurement capabilities with the DeltaTime™ TCSPC accessory. This accessory works in the time domain, mitigating influences from concentration and photobleaching. With its industry-leading 100 MHz system operation, DeltaTime offers rapid TCSPC acquisition rates, enabling fast decay measurements in milliseconds for TCSPC lifetime kinetics of fast reactions. Its crystal-locked timing circuits ensure accuracy. A catalog of over 70 compact pulsed light sources is available, with powerful analysis software supporting 9 fitting models.

Unique TCSPC Benefits

• 40 years of experience in TCSPC innovation.
• Industry-leading true 100 MHz system operation.
• TCSPC decay acquired rapidly.
• TCSPC lifetimes kinetics acquisition up to 1,000 decays per second.
• Lifetimes from <10 ps to seconds.
• True plug-and-play sources requiring no recalibration.
• Unique SpectraLED LED series for high-efficiency phosphorescence measurements.
• Supports TCSPC lifetimes, anisotropy, TRES, and kinetics.
• Provides robust data, independent of concentration and photobleaching.

Descriptions of sample data include lifetimes of rhodamine 6G in methanol showing self-quenching and homo-FRET, and non-extensive distribution fits for 1,8-ANS in bovine serum albumin. A note on sample photobleaching corrupting kinetic data and the FluoroMax's capability for rapid kinetic measurements (as little as every 10 ms) is also included.

Phosphorescence

Measure phosphorescence spectra or lifetimes from microseconds to seconds. The FluoroMax Plus "P" versions feature an optional pulsed xenon flash lamp for lifetime measurements down to 10 µs, suitable for lanthanide-tagged samples or rare earth phosphors.

A description of complex solutions, such as BSA and terbium chloride mixtures, highlights how the pulsed light source can isolate Tb3+ phosphorescence from BSA fluorescence. An inset shows the single wavelength phosphorescence decay of Tb3+.

Enhanced Phosphorescence with SpectraLED LED Sources

HORIBA SpectraLED sources can enhance phosphorescence lifetime measurements. These fixed wavelength sources connect to the sample compartment for direct sample illumination. They offer higher repetition rates for faster acquisitions and variable pulse durations, allowing for measurement of shorter or longer phosphorescence lifetimes.

Extend the Spectral or Lifetime Range of Your FluoroMax Plus

In addition to standard capabilities, the FluoroMax Plus allows adding a second detector on the emission monochromator and a dual grating turret for extended NIR wavelength detection. The second detector is automated with computer control for slit, mirror, and grating selection.

The standard PMT detector covers 185 to 900 nm. A NIR-extended PMT covering 185 to 980 nm is available without needing a second detector port.

Available FluoroMax Plus Second Detectors

• Extended NIR: LN-Cooled InGaAs Detectors (800-2,100 nm), Cooled-NIR PMT (200-1,010 nm), Deep-cooled NIR PMT (950-1,400 nm or 950-1,700 nm).
• ENHANCED TCSPC for Ultra Short Lifetimes: PPD and HPPD Detectors for lifetimes as short as 10 picoseconds.

The unique all-reflective optics design of the FluoroMax Plus focuses all wavelengths to a precise point, avoiding chromatic aberrations common in lens-based systems, which is crucial for extended NIR detection.

A description of a phosphorescence spectrum of singlet oxygen generated by rose bengal in methanol, demonstrating sensitivity with an LN-cooled IGA detector, is provided.

Additional Gratings for Dual-Grating Turret

• 1200 grooves/mm blazed at 500 nm.
• 600 grooves/mm blazed at 1 µm, gold coated for optimized NIR response.
• Other gratings available on request.

Accessories

Microvolumes

Measure ultra-low sample volumes with easy sample recovery. HORIBA's Microsense accessory allows for cuvette-quality steady state or lifetime data from just 1-5 µL of sample. It minimizes sample waste and preserves measurement sensitivity, compatible with UV to NIR measurements.

A description of using the Microsense with 5 µL of AlexaFluor 488 labeled IgG for steady state or TCSPC data is provided.

Microplate High Throughput Accessory

Maximize throughput by automating measurements for a large number of samples. The MicroMax 384 accessory automates measurements on 6 to 384 samples, saving time and money. FluoroMax Plus software includes calibration curve routines for quantification.

Full Spectral Microscopy

FluoroMax Plus systems can be fiber-coupled to fluorescence microscopes for full spectral analysis of spatially varying or small volume samples. This enables measuring spectra from samples as small as 1 µm or a few molecules, and performing localized FRET measurements.

Adding an automated stage and camera allows for creating spatial/spectral maps and performing repetitive QC characterization.

A description of data from real and forged US five dollar bills is given, illustrating how fluorescence imaging can distinguish them spectrally.

Peltier-based Heated/Cooled Cuvette Holders

Provide rapid temperature control for single or multiple cuvette models with magnetic stirring. Precise temperature control is crucial for studying protein folding, solubility, conformation, and phase transitions. Temperature can be varied from -25 to 105° C (optional -40 to 150° C), with software to automate temperature dependence measurements.

A description of thermal unfolding of bovine serum albumin is provided, monitored by tryptophan fluorescence, 1,8-ANS quenching, and scattered light.

Temperature Bath

An alternative for fixing temperature without variable control, offering better precision and long-term stability, though with slower temperature ramping.

Optional LN₂ and He Cryostats

Support various cryostats for greater temperature range and sample type flexibility, offering temperature control down to 4 K.

Economic LN Temperature Dewar Accessory

Cryogenic temperatures enable measurements of fine structure, enhanced phosphorescence, and rare conformations/states. This low-cost accessory allows measurements of liquid or solid samples at 77° K.

A description of spectra of naphthalene dissolved in methanol at room temperature and 77° K is provided, showing vibrational structure and longer wavelength phosphorescence at low temperatures.

Other Accessories

The FluoroMax Plus Series offers a comprehensive line of accessories to extend instrument utility. A partial list includes:

• Solid sample holder for front-face fluorescence of films, powders, pellets, paper, fibers, or slides.
• Thermostated sample holders (2 and 4 position) with magnetic stirring bars.
• Auto-titrator (injector) with dual syringe and valve.
• Hi-tech SFA-20 stopped flow, rapid kinetics accessory.
• Fiber-optic probe for samples not fitting standard chambers.
• Sealed water standard for water Raman S/N verification.
• Emission and Excitation correction factor kits.
• Purge port with quartz windows for nitrogen purging.
• Up conversion laser accessories.
• Reduced volume cells (250 µl) and HPLC flowcells (20 µl).

FluorEssence™ Software

FluorEssence™ software is designed to be simple for occasional users yet powerful for elaborate experiments. It allows for efficient method development, streamlined data collection, analysis, and report generation, full software control of accessories, automation of repetitive experiments via batch mode, and automatic application of instrument calibration parameters.

All experimental parameters are saved with data files for comparison. The software supports conversion of data to answers using tools like OriginPro for processing and data management, with immediate export capabilities.

A diagram illustrates the software's modular structure, including Data Safety, User Interface, Acquisition Control, Timing Electronic Control, Data Display, Power Supply, Data Treatments, Data Acquisition, Communication Protocols, and Instrument Full Control.

Experiment Capabilities & Software Features

The FluoroMax Plus supports a wide range of experiments:

• Spectra: Excitation, emission, and synchronous spectra.
• Kinetics: Time-based intensity measurements.
• 3D Excitation Emission Matrix (EEM) scans.
• Anisotropy measurements (vs. emission, excitation wavelength, time, temperature, single point).
• Single point measurements.
• Phosphorescence measurements (vs. emission/excitation, decay acquisition).
• Lifetime options: Lifetime decay, anisotropy decay (up to 5 exponential), TRES, MCS lifetime (phosphorescence), and Global analysis.

A library of video tutorials is available. Software features include data views in workbook formats, zooming and scaling, contour maps and profiles from 3D plots, peak finding, data fitting (Gaussian, Lorentzian, custom curves), standard arithmetic, 3D perspective, smoothing, deconvolution, excitation/emission correction, interpolation/extrapolation, blank subtraction, normalization, and a PLQY calculator wizard.

FluoroMax Plus Specifications

HORIBA has a policy of continuous product development and reserves the right to amend specifications without prior notice.

Family of Fluorescence Systems

HORIBA offers a range of fluorescence systems:

• Duetta: Affordable Fluorescence and Absorbance Spectrometers.
• Aqualog: Industrial QC/QA.
• FluoroMax Plus: High End Benchtop Fluorometers.
• DeltaFlex: Dedicated TCSPC Lifetime Systems.
• Nanolog: Multichannel Research Fluorometers.
• Fluorolog-QM: Modular Research Fluorometers.

HORIBA Fluorescence: Our Roots Grow Deep

A visual timeline highlights HORIBA's history and acquisitions in fluorescence technology, starting from 1848 (Jobin Yvon) through acquisitions of SPEX, IBH, SLM, PTI, and Photon Technology International, showcasing its long-standing expertise.

Key technologies and applications in HORIBA fluorescence include FRET, EEMs, PLQY, FURA-2, PV, QDot, LRET, GFP, FLIM, and TCSPC.

Contact information and website details are provided for various HORIBA global offices.