A confocal microscopic platform with unique single molecule sensitivity

A modular platform, capable of meeting challenging requirements
The MicroTime 200 contains all optics and electronics needed for recording most aspects of the fluorescence dynamics of samples with single molecule detection sensitivity. The platform’s exceptional sensitivity, flexibility, and unprecedented ease-of-use stem from a unique fusion of miniaturised and highly sophisticated state-of-the-art technologies.

A flexible excitation subsystem, well suited for many samples

The MicroTime 200 features a highly customisable excitation subsystem that can be adapted to requirements of many applications and sample types, such as most organic fluorophores, quantum dots and fluorescent proteins. The subsystem offers:

• Pulsed laser heads covering the spectral range from 375 to 900 nm with pulse widths below 100 picoseconds
• A versatile laser driver (Sepia PDL 828) allows addressing several lasers in parallel to generate complex excitation patterns, which enables advanced techniques such as Pulsed Interleaved
• Excitation (PIE). This technique can help increasing the specificity in single molecule FRET, Fluorescence Correlation Spectroscopy, FLIM, and STED super-resolution imaging.
• Optional support for excitation wavelengths as short as 255 nm, ideal for working with intrinsic fluorescence of aminoacids.
• The possibility for coupling Ti:Sa tunable lasers to enable multi-photon excitation schemes.

Detect your sample’s fluorescence with single photon sensitivity

The MicroTime 200 is specially designed for single molecule studies and thus offers unique optics with vastly reduced light absorption. The detection subsystem offers:

• Up to six individual detection channels, which can each be equipped with a different detector.
• A variety of single photon sensitive detectors, covering different wavelength ranges and detection efficiencies to match the detection requirements of your samples.
• Supports PicoQuant’s PMA Hybrid detectors, which offer high timing precision as well as negligible afterpulsing and dead times (ideal for rapidFLIM).
• A series of optimised SPADs from the PDM Series are also available, offering high detection efficiency.

A confocal microscopy system offering various scanning options

The MicroTime 200 can be equipped with different devices for scanning an image: A galvo scanner for quick image acquisition or piezo scanning devices for maximum flexibility in terms of usable wavelength ranges.

The FLIMbee galvo scanner: combining scan speed flexibility with high position precision
The core of the FLIMbee galvo scanner consists of three high precision oscillating mirrors with excellent linearity, repeatability and low drift.

• FLIMbee achieves extremely stable scanning speeds ranging from very slow to fast while maintaining high positioning precision.
• This high degree of flexibility in speed allows for applications ranging from Phosphorescence Lifetime Imaging (PLIM) to fast fluorescence lifetime measurements using rapidFLIM.
• The FLIMbee’s high positioning precision make it also optimally suited for super-resolution microscopy using STED, enabling imaging down to the single molecule level.

Piezo scanning: high precision with maximum flexibility in useable wavelength range
The MicroTime 200 can be equipped with an objective-scanning solution based on high precision piezo-scanner. This type of scanning is ideally suite for applications which require:

• Excitation light in UV region (255 to 375nm)
• Pixel sizes smaller than 10nm
• Measurements where extreme photon sensitivity matters

Timing electronics with picosecond time resolution and negligible dead time

Time-resolved microscopy requires not only the collection of photons themselves, but also determining their position in time and – for imaging – in space. The ideal technique for that purpose is the Time-Tagged Time-Resolved (TTTR) data acquisition mode developed by PicoQuant, a variation on the classic Time-Correlated Single Photon Counting (TCSPC). This acquisition mode offers the following advantages:

• A fundamental data format allows performing vastly different measurement procedures, such as FLIM, FCS or even coincidence correlation (“antibunching”).
• The TTTR format is supported by all of PicoQuant’s TCSPC electronics.
• The format is also extensively documented and open to the community, simplifiying data transfer to various open source tools.

The MicroTime 200 uses either a MultiHarp 150P or a HydraHarp 400 for data acquisition. For applications requiring extremely high count rates, the MultiHarp 150P is more advantageous. If you aim to push the timing precision to the limit, then HydraHarp 400 is the preferred choice.

MultiHarp 150P HydraHarp 400
Number of detection channels 4, 8, or 16 2. 4, 6, or 8
Dead time < 0.65 ns < 80 ns
Max. sustainable count rate 80 Mcounts per second 40 Mcounts per second
Minimum time bin width 5 ps 1 ps

The SymPhoTime 64 – an intuitive and powerful software for acquisition and analysis of time-resolved microscopy data

SymPhoTime 64 is the system software of the MicroTime 200 that handles all data collection, analysis, and visualisation:

• All data handling in SymPhoTime 64 is fully transparent and maintains all originally collected and derived data in one workspace.
• A comprehensive log file helps to keep track of all measurement and analysis steps.
• Full data export available to standard formats compatible with external analysis tools.

Based on the sophisticated yet easy-to-use data collection and handling processes, the system software supports a multitude of methods, for example:

• Intensity time trace
• Burst analysis
• Lifetime histogramming
• Fluorescence Correlation Spectroscopy (FCS)
• Fluorescence Lifetime Correlation Spectroscopy (FLCS)
• Fluorescence Lifetime Imaging (FLIM)
• Förster Resonance Energy Transfer (FRET)
• Pattern matching analysis
• Fluorescence anisotropy