Multimodal Fluorescence Microscopy and Nanoscopy Platform

This platform consists of a Leica TCS SP8 X system featuring a Leica DMi8 inverted microscope with a motorized stage and various objective lenses (10x to 100x). As exaction source there is a  pulsed 405nm diode laser, a MaiTai DeepSee femtosecond laser (690nm - 1040nm) and a supercontinuum laser (470nm – 670nm) which allows continuously tunable excitation in the visible range coupled with filter-free detection.

For detection, the platform has 2 PMT detectors and four internal hybrid detectors (HyD) which offer high sensitivity and superior signal-to-noise ratio. The system's tandem scanner unites the standard scanner unit with a 8kHz resonant scanner, enabling very fast acquisition. Temperature as well as CO2 control are available for live cell imaging. The system's Lambda square mapping ability allows full spectral analysis of samples and optimization of both the excitation and detection range.

Possibilities of the platform:

  1.  Confocal microscopy TCS SP8 X with resolution below 200nm and 22mm field diameter.

  2. STED nanoscopy: The SP8 STED 3x enables depletion with three different laser lines (592nm CW, 660nm CW, 775nm pulsed), allowing super-resolution imaging both in lateral and axial direction.

  3. 2-Photon microscopy: The setup is equipped with a tunable MaiTai DeepSee femtosecond laser (690nm - 1040nm) for 2-Photon excitation. Two non-descanned HyD detectors for optimal 2-Photon acquisition are included in the setup.

  4. Digital Lightsheet: With minimal adaptation the SP8 can be used as a lightsheet microscope, utilizing Leica’s TwinFlect mirror design for single plane illumination.

  5. TCSPC: The system is coupled with PicoQuant’s time-correlated single photon counting hardware and software enabling various time resolved techniques like FLIM, FRET and FCS.

WEB DEVELOPERS: Coucke, Q., De Ras, M., Vandaele, J.

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