The FluoroMax® Series Spectrofluorometers

The FluoroMax® series constitutes HORIBA’s best-in-class fluorometer performance in a user-friendly, easy and cost-effective benchtop model.

The FluoroMax® range, with its special photon counting and all reflective optics, was the first to introduce the sensitivity of a modular fluorometer to a tabletop fluorescence instrument.

The FluoroMax® is a small spectrofluorometer and yet provides the maximum sensitivity in fluorescence analyses. It also includes features that are not found in a majority of tabletop fluorescence detection systems.

The FluoroMax® series includes the FluoroMax Plus, the next-generation high performance tabletop fluorometer, which delivers more performance with detection of emission spectra out to 1700 nm and time-correlated single-photon counting (TCSPC) lifetime measurements as low as 25 ps.

The FluoroMax® series spectrofluorometers are perfect for quantifying liquid and solid samples.

Video Credit: HORIBA Scientific

Features

  • Photon counting for excellent sensitivity
  • Water Raman signal-to-noise ratio is 10,000:1 (FSD technique)
  • Robust FluorEssence™ software for Windows®
  • Fast scanning capability of up to 80 nm/second
  • Time-correlated single-photon counting (TCSPC) accessory for lifetime determinations
  • Method Files to recall entire experiment parameters
  • Phosphorimeter accessory is available for long-lived luminescence determinations

Hardware

The Source

The xenon source supplying prime UV performance is directed toward the entrance slit of the excitation monochromator with the help of an elliptical mirror. In addition to guaranteeing efficient collection, the reflective surface ensures that all wavelengths are focused on the slit. As a choice of extra, software-selectable, xenon flash lamp can be fixed to allow phosphorescence analyses.

The Slits

The slits themselves can be continuously and bilaterally adjusted from the computer in units of millimeters or bandpass. This sustains instant reproducibility and the highest resolution.

The Excitation Monochromator

The excitation monochromator is an aspheric design that makes sure that the image of the light diffracted by the grating fits via the slit. These are plane and blazed gratings that prevent the two significant drawbacks of the more standard concave holographic gratings—that is, poor polarization performance and insufficient imaging during scans that cast light.

The special wavelength drive scans the grating at rates as high as 80 nm/second. The grating grooves are blazed to offer the highest light in the visible and UV region.

The Reference Detector

Before the excitation light approaches the sample, a photodiode reference detector tracks the intensity as a function of wavelength and time. The photodiode detector has a broader wavelength response when compared to the older, conventional rhodamine-B quantum counter and does not need maintenance.

The Sample Chamber

A spacious sample chamber is available to enable the use of a lengthy list of accessories for unique samples and supports users to use a range of sample schemes.

The Emission Monochromator

All the excellent features of the excitation monochromator have been integrated into the emission monochromator. Gratings are blazed to offer the highest throughput in the visible region.

The Emission Detector

Photon-counting is used by emission detector electronics for maximum low-light-level detection. Photon-counting focuses on signals that emerge from fluorescence emission, neglecting smaller signals that originate in the detector tube (PMT). The more standard technique of analog detection (utilized by lower performance fluorometers) just adds noise and signal collectively, hiding weak emissions.

Besides this, the emission detector housing includes an integral high voltage supply, which is factory set to offer the ideal signal-to-noise ratio.

Computer Control

The complete control of the FluoroMax® originates in users’ PC with HORIBA’s FluorEssence™ software and is sent via a USB link. When the system is turned on, it automatically calibrates and presents itself for novel experiments, or stores routines that are instantly recalled from memory.

Accessories

An extensive range of accessories are applications-oriented, and a majority of them can be interchanged with the FluoroMax® system.

FluorEssence Steady-State and Frequency Domain Software

The FluorEssence™ for Windows® sets a new benchmark for fluorescence software. It combines spectrofluorometer data acquisition instantly to the popular processing power of Origin® software. Simple clicks enable users to choose an accessory, scan type, or restore an entire experimental set-up.

Revolutionize the Way Users Operate their Spectrofluorometer:

  • Thanks to streamlined windows, data acquisition is made intuitive even to casual users
  • Through detector algebra, input signals from various detectors can be assembled into special equations (such as transcendental or arithmetic functions) that generate data in a format customized to users’ applications
  • FluorEssence™ can be learned easily through free video tutorials
  • Units can be changed for wavelength, slit-width and temperature by just clicking the mouse
  • The special Real-Time Control (RTC) window can be selected to improve the run parameters before scanning
  • Blanks and lamp output can be automatically corrected
  • Users can run a range of accessories, such as microwell-plate readers, polarizers, phosphorimeters, autotitrators, temperature baths, etc.

The FluorEssence™ also includes Origin®’s robust processing and data-management tools to ease laboratory operations.

Plotting, data-viewing, processing and management are also available, such as extensive export and import facilities, data management, view manipulation and creation, peak-picking and integration, arithmetic applications, etc.

The FluoroMax® Series Spectrofluorometers

Image Credit: HORIBA Scientific

Software Features

  • Zooming and scaling
  • Data views in workbook formats help keep tables, graphs and notes collectively for every experiment
  • Contour profiles and maps from 3D plots
  • Smoothing
  • Integration or DerivativeFit fluorescence data to Lorentzian, Gaussian and custom curves
  • PLQY calculator wizard (for application with Quanta-Phi accessory)
  • Blank subtraction
  • Deconvolute
  • Standard arithmetic
  • 3D perspective
  • Normalization
  • Interpolation and extrapolation
  • Excitation/emission correction

DAS6 Decay Analysis Software

The decay analysis software from HORIBA Scientific recovers kinetic data (like rotational correlation times and fluorescence lifetime) from luminescence decay data by accommodating the raw decay data to one of a range of kinetic models.

The FluoroMax® Series Spectrofluorometers

Image Credit: HORIBA Scientific

With features like reconvolution of instrument response function (IRF, or 'lamp' profile) and shift iteration, several decay components can be precisely recovered even when the decay data is grossly distorted by the IRF.

DAS6 has been developed to simplify the analysis of time-domain luminescence data and, at the same time, allows the parameters to be further adjusted where required. Analysis modules consist of reconvolution analysis of multi-exponential models as well as analysis of more dedicated fluorescence decay processes as described below.

Multi-Exponential

Analysis carried out for up to 5 exponentials.

Batch Exponential

Batch mode assists datasets consisting of up to 10,000 decays and is perfect for applications where huge amounts of lifetime data are produced, for example, reaction monitoring and lifetime imaging. It features complete reconvolution with up to five exponentials and shift iteration.

Global Exponential

Users can carry out global analyses of up to 100 decay curves and to 5 exponential components

Distribution

There are two techniques for carrying out lifetime distribution analysis. The first technique uses a 'Top Hat' distribution function that enables a single lifetime distribution to be equipped along with an extra discrete lifetime. The second technique is an optional module that will execute a Non-Extensive Decay distribution, which helps model five lifetime distributions.

Exponential Series

The Exponential series helps analyze up to 30 terms. This enables parameterization of otherwise challenging data.

Foerster Energy Transfer

This allows 2D and 3D Foerster-type energy transfer with an exponential component (optional).

Yokoto-Tanimoto Energy Transfer

Yokoto-Tanimoto energy transfer considers diffusion using an optional exponential component.

Micellar Quenching

Micellar quenching kinetics can help find out the surfactant aggregation number.

Exciplex

Exciplex analysis follows the development of an excited state species.

Anisotropy Analysis

Impulse reconvolution of up to five fluorescence decay components and two correlation times is possible. Every model parameter, inclusive of shift, can be set at a predetermined value or improved as part of the parameter fitting.

The use of reconvolution enables precise determination of rotational correlation times and decay times even when the visualized decay is grossly deformed by the duration of the excitation pulse.

DataStation TCSPC Acquisition Software

The FluoroMax® Series Spectrofluorometers

Image Credit: HORIBA Scientific

DataStation is a new measurement environment meant for time-domain fluorescence and is provided with all lifetime systems from HORIBA Scientific.

Lifetime systems are composed of several components and modules, such as monochromators, focusing optics, pulsed light sources, motorized polarizers, MCAs, PMTs, etc. DataStation combines the control of all these components into a single intuitive, mouse-driven Windows environment.

Lifetime systems have the ability to carry out a range of measurement functions, like phosphorescence decay measurement, time-resolved emission scan (TRES), fluorescence decay measurement, time-resolved anisotropy measurements, temperature scans, steady-state spectra, etc.

Today, such measurements are regular through the DataStation. Measurement templates are offered to rapidly set up the instrument for the needed measurement and automated data acquisition is carried out with the help of computer-controlled components.

The acquired data can be stored in a range of formats, such as the .das format, ASCI and HORIBA Jobin Yvon IBH v4.2 printed to users’ Windows printer and copied or exported to the clipboard for manipulation through a third-party application.

The DataStation can be separately bought for use on non-HORIBA Scientific systems, and the company will be glad to add customized functionality and interfacing (for example, to motorized polarizer holders, additional monochromator types, etc).

The DataStation software comes into its own when it is integrated with a FluoroHub.

Functionality

The DataStation software evolves constantly as a new functionality is added in response to customers’ suggestions. The following is a partial list:

Graphical

  • Auto-scaling Y-axis
  • Log or linear Y-axis
  • Reversed time-axis mode (reverses data automatically at the time of reversed-TAC mode measurements)
  • Several mouse zoom modes—x only, y only, x and y, pan, reset
  • Customizable and removable toolbars with helpful hints
  • Real-time display of cursor position, peak channel, peak count and total count
  • Context-sensitive pop-up toolbars
  • Real-time estimated average lifetime during acquisition
  • Intuitive tree navigation for hardware settings and files
  • Tree navigation control window available for FluoroHub control

The FluoroMax® Series Spectrofluorometers

Image Credit: HORIBA Scientific

TCSPC Mode

  • Several traces on screen at the same time
  • Measurement presets—elapsed time or peak count

TRES Mode

  • The instrument can be configured for scan operation by just a few mouse clicks
  • Emission or excitation monochromator can be scanned
  • Manual and automatic data slicing is available
  • Three-dimensional data (counts versus time versus wavelength) can be recorded along with a live 3D progress display and interactive rotation.

Anisotropy Mode

  • G-factor measurement
  • Target difference count

MCA Support

  • EG&G or Perkin Elmer Ortec Trump card
  • Canberra or Tennelec PCA3

Monochromator Support

Polarizer Support

  • PRA polarizers
  • HORIBA Scientific 5000U-02 polarizers

Temperature Control Support

  • HORIBA Jobin Yvon IBH dipstick temperature controller (used as an accessory to HORIBA Scientific 5000 series system only)
  • Cryostat (utilized as an accessory to HORIBA Scientific 5000 series system only)

System Requirements

The below PC specification is needed to run the DataStation software. Measurement functionality relies on the integrated spectrometer hardware. The fundamental operation needs an assisted FluoroHub or MCA card. Users can contact HORIBA Scientific for additional details.

1 Gb RAM, DVD drive, 1 Gb HD space, 1024 x 768 graphics having at least 256 color support, mouse, or other pointing devices. Windows™ XP or later (XP/Vista/7). Windows™ XP is suggested.

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