Picoquant MicroTime 200
Imaging configuration: Confocal for time-resolved fluorescence measurements (FCS, FLIM, FRET)
Widefield
Microscope stand: Olympus IX81, Inverted
Illumination and detection system: Picoquant MicroTime 200
Confocal detection:
Fluorescence excitation: Picoquant picosecond pulsed lasers
Laser line (nm) | Max output power @ 40 MHz (mW) | Pulse duration (ps) |
404 | 3 | 54 |
485 | 5 | 93 (2 laser sources with orthogonal polarization) |
530 | 2 | 83 |
640 | 4 | 63 |
Detection: 2 channels
Detection devices: 2x SPAD from Micro Photon Devices (MPD), PDM series
Single-photon detectors
Quantum efficiency: 50% @ 550 nm
Timing accuracy: < 50ps
Control and acquisition software: Picoquant SymPhoTime
Additional features:
XY mechanical stage
Objective mounted on XYZ piezo stage (2D + PIFOC, +- 100 µm)
Z drift compensation module Olympus ZDC2
Fully equipped for conventional widefield epi-fluorescence imaging (see below for details)
Objectives (from Olympus)
Type | Magnification | Numerical Aperture (NA) | Medium | Contrast | Working distance (mm) |
UPlanSApo | 60x | 1.2 | water | fluo, DIC | 0.28 |
Dichroic mirror for confocal illumination
Laser lines reflected | reference |
530/640 | Z532/635 rpc |
480/640 | 480/640 FITC/Cy5 |
405 | ZT405 rdc |
480/530 | Z480/532 rpc |
Fluorescence emission filters
Emission filter | Type | Reference |
520/35 | Band-pass | FF01-520/35 (x2) |
542/25 | Band-pass | FF01-542/25 |
690/70 | Band-pass | HQ690/70 |
430LP | Long-pass | HQ430LP |
563/9 | Band-pass | HC563/9 |
580/70 | Band-pass | HQ580/70 |
Dichroic beam-splitter for dual-channel detection
Dichroic filter | reference | ||
620 | 620DCXXR (x2) | ||
500 | 500DCXR | ||
532 | Z532 RDC | ||
638 | Z638 RDC |
Confocal pinholes available: 30, 50, 75, 100, 150 µm
Widefield detection:
Fluorescence excitation: High-power lamp Andor AMH-200-F6S
Detection: 1 channel
Detection device: EM-CCD Andor Luca R
Chip size: 1004x1002 pixels
Pixel size: 8x8 µm
Dynamic range: 14 bit
Maximum frame rate: 12 frames/s (full chip)
Quantum efficiency: 65% @ 600 nm
Control and acquisition software: Andor IQ3
Widefield epi-fluorescence filter cubes installed in the microscope
Fluorophore | Excitation filter | Dichroic filter | Emission filter | Filter cube reference |
GFP | 482/18 | 495 | 520/28 | GFP-1828A-OMF-ZERO |
TRITC | 443/22 | 562 | 593/40 | TRITC-B-OMF-ZERO |
Cy5 | 628/40 | 660 | 692/40 | Cy5-4040B-OMF-ZERO |
