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The MicroPL Fluorescence Microscopy Upgrade for Edinburgh Instruments Spectrometers such as the FLS1000 or FS5 allows the study of spectral or time-resolved photoluminescence of samples in the microscopic scale.
An Edinburgh Instruments photoluminescence (PL) spectrometer is converted into a combined spectrometer and microscope system with this user-friendly upgrade.
A wide range of microscope configurations, source coupling and detector options are available enabling both steady state and fluorescence lifetime microscopy, as well as automated maps, using the same setup.
It is also possible to couple your own microscope to the FLS1000 or FS5 for a custom MicroPL upgrade (please get in touch for models supported).
MicroPL is coupled to the spectrometer via liquid light guides or fibres. It is possible to excite the full field of view of the microscope’s objective (widefield excitation), or a specific point in the sample with a laser (point excitation).
The spectrometer software selects the wavelength and bandwidth of the light exciting the sample. This provides much more flexibility than traditional fluorescence microscopy and removes the need for excitation filters.
Point excitation can be performed with either CW or pulsed lasers for spectral or lifetime PL measurements. MicroPL is compatible with Edinburgh Instruments EPL, HPL, and VPL diode lasers. A spot size of ~2 μm (source and objective dependent) is achievable, enabling to resolve PL spectra and lifetimes in the micrometer scale.
Both Time-Correlated Single Photon Counting (TCSPC) and Multi-Channel Scaling (MCS) operating modes are compatible, covering a wide range of photoluminescence lifetimes, from a few ps up to seconds. Non-standard applications such as upconversion PL microscopy are possible with the appropriate choice of filters and laser coupling.
MicroPL can be configured with a computer-controlled XYZ stage for Fluorescence Lifetime Imaging Microscopy (FLIM). The FLIM add-on unlocks special features in the Fluoracle software (FluoracleMap) including control of the stage and advanced analysis options for maps, such as multi-component decay fitting algorithms. No additional software packages are required to obtain and analyse data.
Specifications | |
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Microscope Models | Upright: Nikon NiU, Olympus BX53 Inverted: Nikon Ti2U |
Excitation/Emission Range | 360 nm - 850 nm (std) Can be extended with non-standard UV and NIR objectives |
Excitation Modes | Widefield: Tunable continuous source from spectrometer (steady state) Point: EPL/HPL/VPL pulsed lasers (TCSPC or MCS lifetimes) and CW lasers (steady state) |
Detection Modes | Micro-PL spectroscopy with spectrometer Fluorescence Microscopy with additional camera |
Objective Magnification | Options available from 5X to 100X |
Sample Stage | Manual or PC-controlled XYZ stage with specifications: 75 mm x 50 mm or 130 mm x 85 mm options XY resolution 0.01 µm Z resolution 0.002 µm |
Software | Mapping features in Fluoracle included with PC-controlled XYZ stage |
Accessories | Software-controlled cryostat upgrade |
Specifications | Spectral | Phosphorescence Lifetime | Fluorescence Lifetime |
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Mode of Operation | Single Photon Counting | Time-Resolved Single Photon Counting (Multi Channel Scaling - MCS) | Time-Correlated Single Photon Counting (TCSPC) |
Lifetime range | Milliseconds to hours | 10 ns - 50 s * | 5 ps - 10 µs * |
Sensitivity | >35,000:1 ** | n/a | n/a |
Excitation Sources | |||
Type | 450 W ozone-free xenon arc lamp | Microsecond flashlamp | Picosecond pulsed diode lasers (EPLs, HPLs) and pulsed LEDs (EPLEDs) |
Spectral Range | 230 nm- > 1000 nm | 200 nm - >1000 nm | Discrete wavelengths between 250 nm - 980 nm |
Pulse width | n/a | 1 µs - 2 µs | from 60 ps |
Options | Ozone generating lamp with spectral range 200 nm - >1000 nm | Low to medium repetition rate pulsed lasers. Variable pulse width VPL and VPLEDs | Nanosecond flashlamp 200 nm - >400 nm pulse width <1 ns |
* source and detector dependent ** standard water Raman measurement conditions: Excitation wavelength = 350 nm, Excitation and emission bandwidths = 5 nm, step size = 1 nm, integration time = 1 s, emission wavelength = 397 nm, noise measured at 450 nm and calculation based on the SQRT method |
Monochromator | Specifications |
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Type | Czerny-Turner with 'plug and play' triple grating turret |
Focal Length | 325 mm (double monochromators: 2 x 325 mm) |
Stray Light Rejection | 1:10-5 (single), 1:10-10 (double) |
Gratings | Mounted to triple grating turret |
Accuracy | +/- 0.2 nm * |
Minimum Step Size | 0.01 nm * |
Option | Spectrographs available for operations of CCDs and diode array detectors |
* grating dependent |
Detectors | PMT-900 | PMT-1010 | PMT- 1400/1700 | HS-PMT | MCP-PMT | HS-HPD |
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Spectral Range | <200 nm - 870 nm | <200 nm - 1010 nm | 500 nm - 1700 nm | <230 nm - 850 nm | <200 nm - 850 nm | 220 nm - 870 nm |
Dark Count Rate | <50 cps (-20 °C) | <200 cps (-20 °C) | <20 kcps or <200 kps | <150 cps (0 °C) | <10 cps (-20 °C) | <200 cps (10 °C) |
Reponse Width | 600 ps | 800 ps | 400 ps or 800 ps | 180 ps | <25 ps | 20 ps |
Options | A wide variety of other photomultipliers and analogue detectors are available up to 5,500 nm |