Markterkundung - Vollständig integriertes STED-Mikroskop
Markterkundung - Vollständig integriertes STED-Mikroskop
Mandatory Requirements/Unbedingte Systemanforderungen: - Fully motorized research-grade inverted microscope stand with motorized objective turret, port switchers and filter wheel turret; - Transmitted light: transmitted Koehler illumination light path with high-stability halogen lamp or LED light source for bright field imaging; - Multi-color Widefield LED illumination module for epi-fluorescence imaging; - Fully motorized, high-precision XY scanning stage operating under direct software automation; - Fast Z-nanopositioner (piezo or equivalent) featuring a minimum travel range of 200 μm and sub-nanometer resolution; - Autofocus: Hardware-based automated autofocus compatible with all excitation and depletion lasers for both STED and confocal imaging; - Light Source: At least 7 excitation lasers ranging from 405nm to 685nm covering the spectal range for multi-color imaging (e.g. 405 nm / 440 nm / 488nm / 515 nm /561 nm / 640 nm / 685 nm); - 589 nm (or similar wavelength) STED Depletion laser (min 1 W power, 40 MHz pulsed); - 775 nm (or similar wavelength) STED Depletion laser (min 2.7 W power, 40 MHz pulsed); - All lasers except 405 nm have to be pulsed (40 MHz) and enable Fluorescence Lifetime imaging and gated STED; - 2D/3D STED at a resolution of <75 nm x 75 nm x 75 nm (3D) and <30 nm x 30 nm (2D), respectively: - Fast automated spherical aberration correction for imaging of thick specimen with a sub-millisecond response time; - Spherical aberration correction must be software integrated and allow automated depth-dependent compensation while acquiring 3D volumes with zero hysteresis; - Spherical aberration correction must be available for all high-resolution/high-magnification objectives including the 60-63x oil objective; - Spherical aberration correction must not introduce any chromatic aberrations or field-dependend coma; - High-sensitivity (peak QE at least 50%) Spectral detection: at least five independent spectral detection units freely tunable between 400-800 nm for flexible multi-color imaging; - At least 2 of the spectral detectors have to be area detectors allowing pixel re-assignment for improved z-resolution and imaging of dim samples; - At least 2 of the spectral detectors have to be capable of Fluorescence Lifetime Imaging; - Laser-scanning based transmitted light detector available; - Objectives: High quality, apochromatic objectives ranging from 4x to at least 60x; - 60-63x Oil immersion objective (NA at least 1.42, Working distance at least 0.15 mm) for STED; - 60-63x Silicone-oil immersion objective (NA at least 1.3, Working distance at least 0.3 mm, must have correction collar for correction of spherical aberrations) for live STED imaging of cells and thick specimen; - 60-63x Water immersion objective (NA at least 1.2, Working distance at least 0.28 mm, must have correction collar for correction of spherical aberrations) for STED; - 25x Multi-immersion objective (NA at least 1.0, Working distance at least 1 mm, must have correction collar for adjusting refractive index of medium between 1.41-1.52) for STED; - 20x Air objective (NA at least 0.8, Working distance at least 0.6 mm); - 10x Air objective (NA at least 0.4, Working distance at least 3 mm): - 4x Air objective (NA at least 0.16, Working distance at least 13.0 mm); - Environmental control: Cage incubator for temperature control (~37°C), CO2/O2/N2 control between approx. 0-20% and humidified atmosphere; - Additional high-precision temperature control stage to regulate temperature between -10 °C and +80 °C, +/- 0.1 °C stability; - Integrated control software capable of driving all confocal and super-resolution (STED) hardware components, automation modules, and laser lines, running on a shared database or system architecture; - Control software must provide multiple distinct operating modes or user interfaces tailored to different expertise levels of core facility users/staff (beginner and expert level interfaces); - Control software must support FLIM-based unmixing; - Pixel-wise automatic adaptation of depletion laser power during acquisition based on signal to reduce laser power applied to sampe; - Two high-power computer workstations for acquisition and offline image processing with at least 512 GB of RAM each; - Guaranteed upgradability to support single-molecule localization nanoscopy at a resolution below 5 nm; - The microscope must feature motorized auto-alignment and QC procedures of full beam path including all laser lines and any pinhole to ensure stable performance within a core facility environment.