DeltaPro™ TCSPC Filter Fluorometer

Though it is small and affordable, the DeltaPro™ is a serious performer. Shown is a very short fluorescence lifetime of 93 ps with the data acquired in a mere 100 ms.

 

Erythorosin B in water

 

 

Sample: Erythrosin B in water (µM) 
Source: DeltaDiode-485L @ 100MHz 
Emission: 500nm LP filter 
Time range: 10ns
Acquisition time: 0.1 sec 
<
τ> = 93 ± 4ps 
χ2= 0.96

In accordance with;
†N. Boens et al Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy, Anal. Chem 79 (2007) 2137-2149

Overview

HORIBA Scientific has made a truly affordable Time-Correlated Single Photon Counting (TCSPC) lifetime fluorometer that is extremely accurate, sensitive and fast.

HORIBA Scientific is proud to introduce the DeltaPro™ TCSPC time-resolved filter fluorometer as a low cost dedicated luminescence decay system. The DeltaPro™ has taken the complexity of time-correlated single photon counting (TCSPC) and made it so simple and affordable that any lab can exploit the power of fluorescence dynamics using TCSPC. The system is designed for researchers who demand the ultimate in speed and sensitivity however only need to measure fluorescence or phosphorescence decays at specific wavelengths and do not require spectral capabilities that are provided with the use of an emission monochromator. If you are interested in measuring time-resolved emission spectra (TRES) then you need to consider the HORIBA DeltaFlex™ or the PTI TimeMaster™ series of modular fluorescence lifetime systems. These modular instruments are a good choice for the investigator who has very demanding samples and the money to buy such a system. However the vast majority of researchers are just interested in measuring the fluorescence decays at fixed wavelengths and are happy to save money on the lower cost DeltaPro™ TCSPC system. The DeltaPro™ meets the vast majority of experimental requirements and is extremely easy to use.

Benefits of the DeltaPro™

  • FAST— Acquisition times from one millisecond
  • Sensitive—Single photon counting detection
  • Accurate— Crystal locked timing circuits never require recalibration
  • Measures fluorescence and phosphorescence— Lifetimes from 25 ps to 1 second
  • Compact and easy to use
  • Extensive range of LED’s and diodes for excitation

Limited budget?

If you have a very limited budget and do not require phosphorescence capability then you should also consider our alternative, lower cost, EasyLife™ X fluorescence lifetime system.  Although not TCSPC, the EasyLife™ X can do most of the experiments the DeltaPro™ TCSPC can do for about half the price.

Are you thinking about adding TCSPC fluorescence lifetime components to your own fluorometer?

Do not waste your money on an expensive TCSPC upgrade to your aging fluorometer. The DeltaPro™ TCSPC offers all of the benefits listed above in a new, stand alone, compact instrument. Plus you will have the benefit that your steady state fluorometer and your time resolved DeltaPro™ can both be used at the same time, dramatically increasing your labs productivity.

Proven TCSPC technology with a proud tradition

The DeltaPro™ is the newest low cost TCSPC system from HORIBA Scientific. Consequently there are not many literature citations for the DeltaPro™ itself. However the DeltaPro™ replaces and builds on the platforms of a previous model from HORIBA, called the TemPro which itself was built on more than two decades of TCSPC innovation by IBH. For an updated list of publications citing the use of these HORIBA TCSPC systems click here

Google Scholar

So what are you waiting for?

Give us a call or request a quote for this affordable TCSPC system.

Request Your Quote Today!

 

 

Lifetime Technique

What is TCSPC?

TCSPC (Time-Correlated Single Photon Counting) is the most sensitive and popular method for measuring fluorescence lifetimes. The hardware requirements for a TCSPC system are; pulsed excitation source, single-photon detection module, time digitizer, and decay analysis software. 

TCSPC diagram

The heart of the system is the time digitizer. This can be thought of as a sophisticated stopwatch, where a pulse from the excitation source starts the watch and the detected photons stop it. The stopwatch results are accumulated in a histogram where each column represents a narrow time window after the excitation pulse. As the fluorophores in the sample decay, randomly emitting fluorescence, each photon is detected, timed, counted, and placed in the appropriate histogram column. The process continues (start, stop, count, reset...) until the target quantity of counts is accumulated to reach the required measurement precision. The final histogram represents, in the case of a simple fluorophore, an exponential decay over time. Visual inspection of the data shape provides an intuitive window into the complexity of the sample decay prior to a thorough interpretation using the analysis software.

The main factors to be considered in a TCSPC system are those affecting the temporal resolution (most usefully defined as the shortest lifetime that can be measured). These are typically dominated by the optical pulse width of the excitation source and the transit time spread (TTS, or "jitter") of the single photon detector. Also of importance, considering the photon accumulation process described above, is the efficiency of the system in terms of the timing and processing of photon counts. This relates directly to the repetition rate and the reset time (or deadtime) of the electronics. The DeltaDiode sources operating at up to 100MHz repetition rate, sending a pulse as often as every 10ns, are a perfect match for the DeltaHub, with its ultra low <10ns deadtime. This combination is the most efficient TCSPC system on the market today and allows accurate lifetimes to be acquired in little as 1ms using the DeltaPro™ system. This enables the use of the fluorescence lifetime to monitor kinetic processes. If your measurements require fast acquisition/high throughput, then the DeltaPro™ is the lifetime system for you.

Only one filter based system gives the capability of measuring lifetimes over 11 orders of magnitude (25 ps to 1 s) without the requirement of a costly upgrade. The DeltaPro™ system is the perfect solution for a multi-user facility. 

Advantages of TCSPC with the DeltaPro

  • Single photon sensitivity
  • Ability to measure very weakly emitting samples (femtomolar)
  • Accurate picosecond time resolution
  • Fully digital technique
  • Operates at high repetition rates in concert with ultra low deadtime, leading to short data collection times
  • Compatible with low power excitation sources that are far less likely to cause photobleaching
  • Data has well defined Poisson statistics allowing for more rigorous error analysis.
  • Automatic operation of light sources (full software control)
  • Engineered to offer unbeatable value with standard features including software controlled sample stirring and a phoshorimeter mode
  • No cumbersome PCI cards required

Lifetime Benefits

If you have a steady state fluorometer you need a time-resolved DeltaPro™!

The DeltaPro™ is a compact filter based fluorescence lifetime system that is an excellent companion to any lab that currently uses a steady state fluorometer but does not have access to a fluorescence lifetime system. At a fraction of the cost of a bench-top spectrofluorometer, the DeltaPro™ is extremely easy to use and yet has powerful time-resolved capabilities and decay analysis software.

Time-Resolved Fluorescence (Fluorescence Lifetimes) is an Invaluable Complement to Steady State Fluorescence (Fluorescence Spectra)

If you are currently using a steady state fluorometer for luminescence measurements, but do not have access to a fluorescence lifetime system, you should seriously consider adding the DeltaPro™ to your lab. Fluorescence intensity (steady state) and fluorescence lifetime (time-resolved) measurements are complementary. One must frequently combine results from the steady state fluorescence and the fluorescence lifetimes measurements in order to obtain the most complete information about the molecule(s) of interest. When the first modern day fluorescence lifetime instruments were introduced some thirty years ago, some investigators inherently understood the complementary nature of the fluorescence lifetime technique but it was frankly irrelevant at that time because the cost, size and complexity of those early instruments discouraged all but a relatively few from using this new time-resolved technique. Although instrumentation cost have decreased quite dramatically, as well as the size and complexity of operation, prior to the introduction of the new DeltaPro™, it was still difficult to convince investigators to invest in a fluorescence lifetime instrument.

At a fraction of the cost of a bench-top fluorometer, and because it is just as easy to operate as a fluorometer, the introduction of the DeltaPro™ system has changed people’s attitude towards fluorescence lifetimes as a technique. Now everyone who is doing luminescence measurements can and should put fluorescence lifetimes to good use. By adding time-resolved fluorescence to your research capabilities you will finally be able to fully characterize your fluorescing molecule and molecular systems. For example, you will be able to find out what the rate constants are for the fluorescence emissions and for the non-radiative deactivation of your samples. This information is readily available by combining the lifetime results with the quantum yield values measured from the steady state instrument.

Why fluorescence lifetimes?
Six Important Things You Can’t Do with a Steady State Fluorometer

Differentiate Multiple Structural Domains and Conformations

HSA excited at 295nm, with the decay of the tryptophan modeled using NED distribution analysis

If you want to characterize a molecule’s interactions with the surrounding environment, the steady-state measurement alone can provide a fluorescence spectrum, fluorescence quantum yield or anisotropy value, however most of this information is scrambled together, as the measured parameters are time averages and the information about specific processes is lost. This lost information becomes especially important when fluorescent molecules are used as probes to study complex systems, such as proteins, nucleic acids, quantum dots, membranes, polymers, surfactants (micelles) etc. These systems frequently exhibit multiple structural domains and conformations. The fluorescence lifetime decay curve will reveal this information by detecting multiple fluorescence lifetimes, which cannot be gleaned with a steady-state measurement where all of this information is totally obscured. The DeltaPro™ software even includes extremely powerful decay analysis software including MEM and our new NED routines.

Study Protein Conformation Dynamics

Phosphorescence decay of HSA excited using a SpectraLED-295, showing the change in average lifetime (calculated from a 3 exponential fit) with temperature

A very powerful application for a time-resolved fluorescence instrument is the study of multiple conformational states of a protein. Consider a simple case of a protein containing one Tryptophan (Trp) residue (e.g. human serum albumin HSA). With a steady state instrument all you can measure is a typical Trp spectrum reflecting no particular information about the protein, except that it contains Trp. However, if you measure the fluorescence decay, you’ll find that this single Trp residue has 3 different discrete fluorescence lifetimes!  By monitoring changes in them and their contribution to the overall emission conformational changes can be inferred. To uncover larger protein motions it is advantageous to use phosphorescence, as this timescale is more applicable so some domain movements. Again, changes in the phosphorescence lifetime and its component contributions can clearly show changes in conformation.

From changes in the lifetimes and their contributions you can immediately see changes in conformational states.

Binding Efficiency (Bound versus Unbound) of Fluorescence Probes

Kinetic TCSPC measurement showing diphenylheptanoid binding to serum albumin. Measured using a DeltaDiode laser with a 100MHz excitation rate and an acquisition time of 2ms/pt. The increase in the lifetime (fitted to a monexponential model) upon binding to serum albumin is clearly seen.

A steady state experiment can reveal binding between a fluorescent probe and a protein. Normally, the fluorescence intensity will change as a result of binding; it will either decrease or increase, depending on the nature of the probe. The information you get is very general. You detected that binding has occurred or not and that is all. However using lifetimes you can uncover different lifetimes, one for the bound and the other for the unbound probe, as well as their relative contributions (pre-exponential factors) to the overall decay. From the lifetime measurement you now know relative populations of bound and unbound probes (i.e. we know the efficiency of binding). Making use of high repetition rate excitation sources, coupled with very low deadtime electronics, enables the fluorescence lifetime to also be used to follow the actual binding kinetics, as measurement times down to 1ms are possible.

Trp Localization in Protein via Fluorescence Quenching

One of the major tools of fluorescence is studying quenching of fluorophores by adding quencher molecules. For example, tryptophan residues in a protein can be quenched by acrylamide or iodide ions. A steady state experiment can show the decrease of fluorescence intensity as the quencher is added, and hence that quenching has occurred, but it cannot tell you if it was dynamic or static quenching. A fluorescence lifetime experiment however will detect more than one lifetime due to different sites that Trp may occupy in the protein. Furthermore the fluorescence decay will provide the quencher effect on each lifetime, so you can get information about localization of each type of the Trp residues (e.g. are they surface exposed or buried inside the protein).

Time-Resolved FRET Checker: “Is that really FRET you are measuring?”

The Förster Resonance Energy Transfer (FRET) technique has become a very powerful and wide-spread experimental tool for studying molecular binding. It is equally popular on the cellular level with fluorescence microscopes as well as in molecular solutions in cuvettes. However most investigators are using steady state techniques to observe and quantitate the ratio of fluorescence intensities of the acceptor and donor wavelengths. This has lead to a number of false conclusions and an increased awareness that the time-resolved technique is really the only way to be certain that you are actually measuring FRET. 

Having a time-resolved fluorescence system is essential because the actual mechanism of fluorescence quenching in general cannot be revealed by the steady state experiment at all. There are two mechanisms that lead to quenching. The first is collisional (or dynamic) quenching, where the excited fluorophore and quencher collide and diffuse apart. The second is static quenching, where fluorophore in the ground state forms a non-fluorescent complex with quencher. In both cases the steady state experiment will show intensity decrease as more and more quencher is added. In the case of collisional (dynamic) quenching the lifetime measurement will show the lifetime decrease as more quencher is added. However, in the case of static quenching there will be no change in the lifetime at all. Discerning between the two mechanisms is critically important when one is looking to study Förster Resonance Energy Transfer (FRET). Only the time-resolved technique can prove that a ‘FRET-like’ behavior is not caused by static quenching. Only a lifetime experiment can rule it out.

Hardware

DeltaPro TCSPC filter fluorometer

Shown beside the laptop is the DeltaPro sample compartment (middle), DeltaHub TCSPC interface (below right), NanoLED control box (above right), and a NanoLED (left) connected to the sample compartment. Not visible is the PPD detector connected to the back of the sample compartment.

The DeltaPro™ is a compact filter-based TCSPC fluorescence lifetime system that comes complete with PPD detection technology and software. There are number of light source options to choose from, as well as a number of accessories, such as sample handling and polarizers. At a minimum, all you need is one pulsed light source and a computer, or laptop, with a single USB port available. The system is so easy to use that it does not require installation.

Included with the System


The DeltaPro™ is a very simple system requiring no installation. It is shipped with the following components which are easily assembled by the user.

  •  Sample chamber with lid, containing a sample holder (with Integral stirrer and temperature sensor) and with two optical ports for the attachment of the light source and detector.
  • DeltaHub™ TCSPC electronics module
  • Light source(s) (NanoLED / DeltaDiode™ / SpectraLED) with appropriate controller (as specified with order)
  • PPD™ picosecond photon detector
  • Optional power supply (DPS-1) for longer wavelength detector versions (PPD-850, PPD-900) if ordered
  • DeltaHub™ User Guide containing CD-ROM with software, drivers and instructions
  • Relevant manuals
  • Accessories specified at time of order

Three Types of pulsed Light Sources to Choose From

There is a comprehensive range of pulsed illuminators to choose from for use with the DeltaPro™. For fluorescence lifetimes you can choose from either the NanoLED™ or the DeltaDiode™ series of sources. Each series includes a mix of pulsed LED’s and lasers, with the fundamental difference the maximum repetition rate that they can operate at (1 MHz for the NanoLED™ and up to 100 MHz for the DeltaDiode). Generally speaking, the higher the repetition rate the shorter the measurement time. Each series has its own dedicated driver interface. So for example a DeltaDiode driver cannot be used with a NanoLED illuminator and vice versa.

For longer lived phosphorescence lifetimes the DeltaPro™ uses the SpectraLED series of software adjustable LED illuminators.

 

Type

Primary Benefit

Repetition Rate

Lifetime Range

Available Wavelengths (nm)

NanoLED™

LED or Laser

Lowest Cost

10 KHz to 1 MHz

30 ps to 1 µs

250 to 1,310* nm

DeltaDiode™

LED or Laser

Fastest

10 KHz to 100 MHz

25 ps to 1 µs

250 to 1,310* nm

SpectraLED™

LED

Phosphorescence

0.1 to 3 KHz

1 µs to 1 s

265 to 1,275* nm


* Note that although these light sources are available out to long NIR wavelengths, the DeltaPro™ is only able to detect to 920 nm with the optional red extended PPD detector. If you need to measure NIR fluorescence lifetimes please look into our high end modular fluorescence and phosphorescence lifetime systems available from the HORIBA Scientific Fluorescence Group which now also includes the complete line of PTI Fluorescence Systems.

 

NanoLED™ Light Sources

NanoLED

The NanoLED is a novel and economical series of light sources that utilize laser diode and LED technology to generate short optical pulses over a wide range of repetition rates and wavelengths.  Optical pulses as short as 70ps can be generated at repetition rates up to 1MHz. All NanoLED sources are controlled with a NanoLED driver interface.

Each NanoLED is designed to be used at a specific wavelength and has a pulse width of 200 ps to 1.6 ns (LED dependent). For the DeltaPro NanoLED sources can be selected from 250 to 830 nm. 

Current list of available NanoLED sources for the DeltaPro™

Model

Peak wavelength (nominal)

Source type

Pulse width typical

NanoLED-250

250 ±10nm

LED

<1.2ns

NanoLED-260

260 ±10nm

LED

<1.2ns

NanoLED-265

265 ±10nm

LED

<1.2ns

NanoLED-270

270 ±10nm

LED

<1.2ns

NanoLED-280

280 ±10nm

LED

<1.2ns

NanoLED-290

290 ±10nm

LED

<1.2ns

NanoLED-295

295 ±10nm

LED

<1.2ns

NanoLED-300

300 ±10nm

LED

<1.2ns

NanoLED-310

310 ±10nm

LED

<1.2ns

NanoLED-320

320 ±10nm

LED

<1.2ns

NanoLED-330

330 ±10nm

LED

<1.2ns

NanoLED-340

340 ±10nm

LED

<1.2ns

NanoLED-350

350 ±10nm

LED

<1.2ns

NanoLED-360

360 ±10nm

LED

<1.2ns

NanoLED-370

370 ±10nm

LED

<1.2ns

NanoLED-375L

375 ±10nm

Diode Laser

<200ps

NanoLED-390

390 ±10nm

LED

<1.3ns

NanoLED-395L

395 ±10nm

Diode Laser

<200ps

Model

Peak wavelength (nominal)

Source type

Pulse width typical

NanoLED-405L

405 ±10nm

Diode Laser

<200ps

NanoLED-415L

415 ±10nm

Diode Laser

<200ps

NanoLED-425L

425 ±10nm

Diode Laser

<200ps

NanoLED-440L

440 ±10nm

Diode Laser

<200ps

NanoLED-455

455 ±10nm

LED

<1.3ns

NanoLED-470L

470 ±10nm

Diode Laser

<200ps

NanoLED-485L

485 ±10nm

Diode Laser

<200ps

NanoLED-510L

510 ±10nm

Diode Laser

<200ps

NanoLED-570

570 ±10nm

LED

<1.5ns

NanoLED-590

590 ±10nm

LED

<1.5ns

NanoLED-605

605 ±10nm

LED

<1.5ns

NanoLED-625

625 ±10nm

LED

<1.4ns

NanoLED-635L

635 ±10nm

Diode Laser

<200ps

NanoLED-650L

650 ±10nm

Diode Laser

<200ps

NanoLED-670L

670 ±10nm

Diode Laser

<200ps

NanoLED-740

740 ± 20nm

LED

<1.6ns

NanoLED-785L

85 ± 20nm

Diode Laser

<200ps

NanoLED-830L

830 ± 20nm

Diode Laser

<200ps

 

New NanoLEDs heads are continually under development — contact HORIBA Scientific for other wavelengths.

DeltaDiode™ Light Sources

DeltaDiode

The world's first 100MHz picosecond diode source with true plug-and-play interchangeable heads and USB interface!

DeltaDiode sources utilize laser diode and LED technology to generate short optical pulses over a very wide range of repetition rates and wavelengths. Optical pulses as short as 40 ps can be generated at repetition rates up to 100MHz. State of the art features such as hot-swap heads, USB control and full software integration mean the DeltaDiode delivers the highest level of performance in the most user-friendly package available today.

All DeltaDiode sources are controlled with a DeltaDiode driver interface.
DeltaDiode heads are switched using a single connection, with optimized settings stored in each head to enable the DeltaDiode software to "know" which head is connected.

Each DeltaDiode is designed to be used at a specific wavelength and has a pulse width of 40 to 1.5 ns (diode dependent). DeltaDiode sources can be selected from 250 to 830 nm for use with the DeltaPro. 

Current list of available DeltaDiode sources for the DeltaPro™

Model

Peak wavelength (nominal)

Source type

Pulse width (Typical)

Pulse width (Max)

Peak power

Average power

Max repetition rate

DeltaDiode-250

250nm ± 10nm

LED with active temp control

750ps

1.0ns

240mW

4µW

20MHz

DeltaDiode-260

260nm ± 10nm

LED with active temp control

750ps

1.0ns

290mW

5µW

20MHz

DeltaDiode-270

270nm ± 10nm

LED with active temp control

800ps

1.2ns

520mW

10µW

20MHz

DeltaDiode-280

280nm ± 10nm

LED with active temp control

950ps

1.5ns

170mW

10µW

20MHz

DeltaDiode-290

290nm ± 10nm

LED with active temp control

800ps

1.2ns

520mW

10µW

20MHz

DeltaDiode-300

300nm ± 10nm

LED with active temp control

800ps

1.2ns

260mW

5µW

20MHz

DeltaDiode-310

830nm ± 10nm

LED with active temp control

800ps

1.2ns

260mW

5µW

20MHz

DeltaDiode-320

320nm ± 10nm

LED with active temp control

800ps

1.2ns

260mW

5µW

20MHz

DeltaDiode-330

330nm ± 10nm

LED with active temp control

800ps

1.5ns

100mW

2µW

20MHz

DeltaDiode-340

340nm ± 10nm

LED with active temp control

750ps

1.0ns

120mW

2µW

20MHz

DeltaDiode-350

350nm ± 10nm

LED with active temp control

800ps

1.0ns

120mW

2µW

20MHz

DeltaDiode-360

360nm ± 10nm

LED with active temp control

800ps

1.0ns

120mW

2µW

20MHz

DeltaDiode-370

372nm ± 10nm

LED

800ps

1.2ns

100mW

2µW

20MHz

DeltaDiode-375L

375 ±7nm

Laser Diode with active temp control

45ps

70ps

300mW

1.8mW

100MHz

DeltaDiode-395L

395 ±8nm

Laser Diode with active temp control

70ps

90ps

400mW

3.0mW

100MHz

DeltaDiode-405L

405 ±8nm

Laser Diode with active temp control

45ps

70ps

300mW

1.4mW

100MHz

DeltaDiode-415L

415 ±8nm

Laser Diode with active temp control

70ps

90ps

250mW

1.7mW

100MHz

DeltaDiode-425L

425 ±8nm

Laser Diode with active temp control

70ps

70ps

230mW

1.6mW

100MHz

DeltaDiode-440L

440 ±9nm

Laser Diode with active temp control

60ps

70ps

400mW

3.0mW

100MHz

DeltaDiode-450L

450 ±10nm

Laser Diode with active temp control

80ps

120ps

250mW

3.0mW

100MHz

DeltaDiode-470L

472 ±7nm

Laser Diode with active temp control

65ps

90ps

160mW

1.5mW

100MHz

DeltaDiode-485L

485 ±10nm

Laser Diode with active temp control

80ps

95ps

300mW

3.0mW

100MHz

DeltaDiode-510L

508 ±10nm

Laser Diode with active temp control

110ps

140ps

100mW

1.4mW

100MHz

COMING SOON – laser modules at 532 nm, 560 nm and 590 nm

DeltaDiode-635L

635 ±10nm

Laser Diode with active temp control

60ps

80ps

300mW

2.0mW

100MHz

DeltaDiode-650L

650 ±15nm

Laser Diode with active temp control

70ps

90ps

60mW

0.4mW

80MHz

DeltaDiode-670L

670 ±10nm

Laser Diode with active temp control

75ps

90ps

30mW

0.2mW

100MHz

DeltaDiode-730L

730 ±10nm

Laser Diode with active temp control

60ps

70ps

500mW

3.0mW

100MHz

DeltaDiode-785L

785 ± 10nm

Laser Diode with active temp control

60ps

80ps

250mW

2mW

100MHz

DeltaDiode-830L

830nm ± 10nm

Laser Diode with active temp control

50ps

70ps

200mW

0.6mW

100MHz

 

New laser heads are continually under development — contact HORIBA Scientific for other wavelengths

SpectraLED Light Sources

SpectraLED

The SpectraLED is a novel light source designed specifically for the measurement of phosphorescence lifetimes. These sources are based on LED technology and the illumination wavelengths range from the UV to the NIR.

The addition of a SpectraLED source to the DeltaPro TCSPC system enables phosphorimeter capability and the measurement of luminescence lifetimes ranging from picoseconds to seconds in one compact system.

The SpectraLED is a modern approach to measuring longer lived luminescence decays. Traditionally, phosphorescence lifetime measurements are excited using a xenon flash lamp. Xenon lamps are broadband sources and offer complete wavelength coverage from the deep UV to the NIR region. For applications where continuous wavelength tunability is not required, SpectraLED sources are a convenient alternative with the following advantages:

SpectraLED Benefits

DeltaPro TCSPC check!

Operation at higher repetition rates not limited by capacitor charging times

DeltaPro TCSPC check!

Software control of pulse duration and repetition rate to optimally excite the sample under investigation

DeltaPro TCSPC check!

No afterglow, permitting easier interpretation of lifetimes shorter than 100 µs

DeltaPro TCSPC check!

Silent operation

 

Each SpectraLED is designed to be used for a specific wavelength range and has a variable pulse width from 100 ns to milliseconds, which is controlled by the DeltaPro acquisition software. SpectraLED’s can be selected from 265 to 835 nm for use with the DeltaPro and operate at repetition rates from 0.1 to 3 KHz depending on the measurement range.

SpectraLED sources do not require a special driver. They are software driven directly from the DeltaHub TCSPC interface.

Current list of available SpectraLED sources for the DeltaPro™

Source

Peak wavelength (nm)

Typical Spectral FWHM (nm)

S-265

265 +/- 10nm

10

S-280

280 +/- 10nm

10

S-290

290 +/- 10nm

10

S-295

295 +/- 10nm

10

S-310

310 +/- 10nm

10

S-330

330 +/- 10nm

10

S-340

340 +/- 10nm

10

S-350

350 +/- 10nm

14

S-370

370 +/- 10nm

15

S-390

390 +/- 10nm

20

Source

Peak wavelength (nm)

Typical Spectral FWHM (nm)

S-415

415 +/- 10nm

20

S-460

460 +/- 10nm

30

S-495

495 +/- 10nm

30

S-535

535 +/- 20nm

30

S-560

560 +/- 10nm

15

S-590

590 +/- 10nm

15

S-605

605 +/- 15nm

20

S-625

625 +/- 10nm

25

S-740

740 +/- 10nm

25

S-830

830 +/- 10nm

50

 

New laser heads are continually under development — contact HORIBA Scientific for other wavelengths

Customer Supplied Light Sources

In addition to selecting from the current list of DeltaDiode, NanoLED and SpectraLED light sources, it is possible to make use of other customer supplied pulsed light sources with the DeltaPro™. Consult with HORIBA for use of your pulsed illuminator with the DeltaPro™ TCSPC system.  

 

Software

Don't let the low price of the DeltaPro™ fool you. This low cost system comes with the same world-leading TCSPC decay acquisition and analysis software included with the most advanced HORIBA TCSPC research systems.

World-Leading Decay Analysis Software

HORIBA Scientific decay analysis software recovers kinetic information (such as fluorescence lifetime and rotational correlation times) from luminescence decay data by fitting the raw decay data to one of a selection of kinetic models. Features such as shift iteration and reconvolution of the instrument response function IRF, or prompt, allow the accurate recovery of multiple decay components, even when the decay data is grossly distorted by the IRF.

DAS6, the latest version of our decay analysis software, uses proprietary hybrid grid-search algorithms that rigorously avoid false minima in chi-squared and erroneous decay constants. The license comprises the main application workspace, and allows reconvolution analysis with up to five exponentials. It also includes anisotropy, batch analysis, lifetime distribution, micellar quenching and energy transfer.

Functionality

HORIBA Scientific DAS6 is designed to streamline the analysis of time-domain luminescence data, while still allowing the opportunity for fine-tuning of parameters where necessary. The DAS6 lifetime fitting module contains reconvolution analysis with up to 5 exponentials, as well the following more advanced fitting modules.

Batch mode

Batch mode analyzes datasets containing up to 10,000 decays and is ideal for applications where large amounts of lifetime data are generated e.g. lifetime imaging and reaction monitoring. This module features full reconvolution with up to five exponentials and shift iteration. Parameters can be held common between datasets, for ‘quasi-global analysis’. All data and results are stored in a single data file and results are automatically tabulated for use in external spreadsheet applications.

Global analysis

Full Global analysis of up to 5 exponential components and 100 decay curves.

Lifetime distribution

We offer three distributions analysis models to choose from.

The Top-hat model fits a distribution of decay times and distribution plus additional exponential.

The Maximum Entropy Method (MEM) is designed to recover lifetime distributions without any a priori assumptions about their shapes. This method uses a series of exponentials (up to 200 terms) as a probe function with fixed, logarithmically-spaced lifetimes and variable pre-exponentials.

Our Non-extensive decay (NED) analysis is based on the gamma function distribution, which is the most probable form of distribution for positive lifetimes. Unlike “model free” analyses, it starts with a defined number of distributions. This constraint means that it is less likely to produce artifacts and is a simpler method to fit both complex as well as purely discrete exponential decays.

 

HSA excited at 295nm, with the decay of the tryptophan modelled using NED distribution analysis

HSA excited at 295nm, with the decay of the tryptophan modeled using NED distribution analysis

Fit to exponential series

Exponential series analysis of up to 30 terms.

Time-resolved anisotropy

Impulse reconvolution of up to 2 correlation times and 5 fluorescence decay components. Each model parameter, including shift, can be fixed at a pre-determined value or optimized as part of the parameter fitting. The use of reconvolution allows accurate determination of decay times and rotational correlation times even when the observed decay is grossly distorted by the duration of the excitation pulse.

FRET energy transfer

2-D and 3-D FRET-type energy transfer with optional additional exponential component.

Yokota-Tanimoto energy transfer

Yokoto-Tanimoto energy transfer with optional additional exponential component.

Micellar quenching

Micellar quenching kinetics.

Unique HORIBA Software Benefits

  • Proven fit algorithms with reconvolution of IRF (prompt)
  • Shift iteration - compensates for wavelength and quantization shifts
  • Unique grid-search algorithm for superior chi-squared minimization (fewer false minima than general purpose Marquardt)
  • "Recommend" feature automatically fills in initial estimates for fit parameters
  • Simplified file handling, .DAS file format stores data and analyzes in one file
  • Analyze up to 10,000 decays

Computer Requirements

A computer or laptop with WindowsXP or Windows 7 (32 or 64 bit), English language, with one available USB 2.0 port is all that is required to run the DeltaPro™ and DAS6 software.

 

Specifications

Specifications

Minimum Lifetime

25 ps with DeltaDiode laser diode source*

Shortest acquisition time

1 millisecond* (requires DeltaDiode laser diode source)

Repetition rates

10 kHz – 1 MHz with NanoLED 
10 kHz – 100 MHz with DeltaDiode
0.1 Hz – 3 kHz with SpectraLED

Diode controller

DeltaPro-NL: NanoLED and SpectraLED
DeltaPro-DD: DeltaDiode and SpectraLED

Prompt (IRF) 

< 200 ps (with 405nm laser diode)

Deadtime

10 ns

Time ranges

10 ns - 11s (with DeltaDiode laser diode)

From 100 ns with NanoLED

Excitation wavelengths
(source dependent)

250 to 830 nm (NanoLED) 
250 to 830 nm (DeltaDiode) 
265 to  830 (SpectraLED)

Emission wavelength selection

Interchangeable filters (not included) – 50 mm

Detector response

230 to 700 nm (standard)
230 to 850 nm (optional)
230 to 920 nm (optional)

PC interface

USB 2.0.  PC not included.  Requires Windows XP or Windows 7, 32/64-bit English language ver.

System footprint

75cm x 45cm x 25cm (W x D x H) excluding PC

*dependent on sample and system configuration

 

Accessories

Filters

DF-mount

Filter and  mount for NanoLED sources

DF-265-25-20

Filter and mount for NanoLED-265 source, 25nm bandwidth, 20% transmittance

DF-280-25-20

Filter and mount for NanoLED-280 source, 25nm bandwidth, 20% transmittance

DF-295-25-20

Filter and mount for NanoLED-295

DF-310-10-15

Filter and mount for NanoLED-310 source, 10nm bandwidth, 15% transmittance

DF-330-10-20

Filter and mount for NanoLED-330 source, 10nm bandwidth, 20% transmittance

DF-340-10-25

Filter and mount for NanoLED-340 source, 10nm bandwidth, 25% transmittance

DF-360-10-25

Filter and mount for NanoLED-360 source, 10nm bandwidth, 25% transmittance

DF-370-10-25

Filter and mount for NanoLED-370 source, 10nm bandwidth, 25% transmittance

DF-375-6-90

Filter and mount for NanoLED-375L source, 6nm bandwidth, 90% transmittance

DF-390-10-30

Filter and mount for NanoLED-390 source, 10nm bandwidth, 30% transmittance

DF-405-10-90

Filter and mount for NanoLED-405L source, 10nm bandwidth, 90% transmittance

DF-440-10-90

Filter and mount for NanoLED-440L source, 10nm bandwidth, 90% transmittance

DF-450-10-45

Filter and mount for NanoLED-450 source, 10nm bandwidth, 45% transmittance

DF-460-10-45

Filter and mount for NanoLED-460 source, 10nm bandwidth, 45% transmittance

DF-470-10-90

Filter and mount for NanoLED-470L source, 10nm bandwidth, 90% transmittance

DF-490-10-45

Filter and mount for NanoLED-495 source, 10nm bandwidth, 45% transmittance

DF-560-10-50

Filter and mount for NanoLED-560 source, 10nm bandwidth, 50% transmittance

DF-590-10-50

Filter and mount for NanoLED-590 source, 10m bandwidth, 50% transmittance

DF-610-10-50

Filter and mount for NanoLED-605 source, 10nm bandwidth, 50% transmittance

DF-620-10-50

Filter and mount for NanoLED-625 source, 10nm bandwidth, 50% transmittance

DF-640-8-90

Filter and mount for NanoLED-635L source, 8nm bandwidth, 90% transmittance

DF-650-13-90

Filter and mount for NanoLED-650L source, 13 nm bandwidth, 90% transmittance

DF-670-10-55

Filter and mount for NanoLED-670L source, 10nm bandwidth, 55% transmittance

DF-730-10-50

Filter and mount for NanoLED-730 source, 10nm bandwidth, 50% transmittance

DF-740-10-50

Filter and mount for NanoLED-740 source, 10nm bandwidth, 50% transmittance

DF-830-10-50

Filter and mount for NanoLED-830L source, 10nm bandwidth, 50% transmittance


KV set of bandpass filters: KV370, KV399, KV450, KV500, KV550
Neutral density filters: (0.3 OD ND, 1.0 OD ND, 2.0 OD ND) 
Bandpass filters are available at most wavelengths. 

Polarizers

The DeltaPro offers optional manual polarizers.

cuvettes

Polarizers are used to separate the light into vertical and horizontal components. This allows allow the DeltaPro to measure time-resolved fluorescence anisotropy which allows you to estimate the rate of rotational diffusion and the actual size of the macromolecule a fluorescent probe is attached to.

Optional sample holders

cuvettes

Shown from left to right: Standard cuvette holder, optional solid sample holder, and the optional front face cuvette holder

  • Solid sample holder
  • Front face water jacketed cuvette holder

 

 

 

 

Data

Though it is small and affordable the DeltaPro™ is a serious performer

Below is a sampling of data acquired with the DeltaPro TCSPC system.

Fast data acquisition (1ms)

Fast data acquisition (1ms)

Sample: Erythrosin B in MeOH (µM) 
Source: DeltaDiode-485L @ 100 MHz
Emission: 500 nm LP filter
Time range:  10 ns
τ = 0.45 ± 0.06 ns
χ2 = 0.96

† In accordance with N. Boens et al Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy, Anal. Chem 79 (2007) 2137-2149

Acquisition of short lifetime

Acquisition of short lifetime

Sample: Erythrosin B in water (µM) 
Source: DeltaDiode-485L @ 100 MHz 
Emission: 500 nm LP filter 
Time range: 10 ns
Acquisition time: 0.1 sec 
<τ> = 93 ± 4ps 
χ2 = 0.96

† In accordance with Boens et al Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy, Anal. Chem 79 (2007) 2137-2149

Short anisotropy decay using reconvolution

Short anisotropy decay using reconvolution

Sample: Coumarin 6 in EG/MeOH mixture (µM) 
Source: DeltaDiode-425L
Emission:  500 nm LP filter
Time range:  100 ns
Anisotropy decay acquisition time: 80 seconds
τR = 112 ps
R0 = 0.32
χ2 = 1.11

Lifetime of plant leaf

Lifetime of plant leaf

Source: DeltaDiode-485L
Emission:  550 nm LP filter
Time range:  40 ns
Acquisition time: 5 seconds
<τ> = 681 ps
χ2 = 1.12

Lifetime of Ru derivative

Lifetime of Ru derivative

Source: DeltaDiode-440L
Emission:  600 nm LP filter
Time range:  6.5 µs
<τ> = 0.41 µs 
χ2 = 1.05

Lanthanide in a glass matrix

Lanthanide in a glass matrix

Source: SpectraLED-370
Emission:  550 nm LP filter
Time range:  44 ms
Acquisition time: 6 seconds
<τ> = 2.7 ms
χ2 = 1.13

Room temperature protein phosphorescence

Room temperature protein phosphorescence

Sample: HSA (µM) 
Source: SpectraLED-295
Emission:  450 nm LP filter
Time range:  44 ms
Acquisition time: 470 seconds
<τ> = 1.4 ms
χ2 = 1.03

K. Sagoo, R. Hirsch, P. Johnston, D. McLoskey and G. Hungerford, 2014. Pre-denaturing transitions in human
serum albumin probed using time-resolved phosphorescence.
Spectrochim. Acta A. 124, 611-617.

Kinetic TCSPC measurement

Kinetic TCSPC measurement

Kinetic TCSPC measurement showing diphenylheptanoid binding to serum albumin. Measured using a DeltaDiode laser with a 100MHz excitation rate and an acquisition time of 2ms/pt. The increase in the lifetime (fitted to a monexponential model) upon binding to serum albumin is clearly seen

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