|Leica TCS SP8||Leica TCS SP8 SMD||Leica TCS SP8 DLS|
|Use:||Standard confocal microscope for fixed samples,
short live-cell experiments, FRAP, FLIP and FRET.
|Special confocal microscope
for fixed samples, short/long live-cell experiments, FLIM/FRET, FRAP, FLIP
|Special confocal microscope for fixed samples and live species, spheroids and cleared tissue.|
|Access:||Core facility, please choose this microscope first if you are planning your experiments and you do not need the special SP8 SMD or DLS features.||Core facility, please choose this microscope only if the SP8 X is fully booked, operators can ask you to shift bookings to another date if special experiments of other users need the SP8 SMD.||Core facility, please choose this microscope only if the SP8 X or SMD is fully booked, operators can ask you to shift bookings to an other date if special experiments (Lightsheet) of other users need the SP8 DLS.|
|Stand:||Leica DMI 6000 with adaptive
|Leica DMI 6000 with adaptive
focus with touch controlller
|Leica DMi8 CS (no adaptive focus)|
|Stage:||Motorized XY-Stage and SuperZ Galvo|
|Incubator:||No||Case Incubator, 37°,
CO2 (standard on), so lean back the transmitted light arm only
for changing samples and xy-stage initialization.
|Objectives:||10x/0.40 IMM CS 20x/0.75 IMM Corr CS2 40x/1.30
Oil CS2 40x/1.10 Water MotCorr CS2 63x/1.40 Oil CS2
|20x/0.75 IMM Corr CS2 40x/1.30
Oil CS2 63x/1.10 Water MotCorr CS2 63x/1.40 Oil CS2
|10x/0.5 Air CS2 40x/1.30 Oil CS2 63x/1.40 Oil CS2|
|All objectives are HC Plan Apochromats. IMM = Glycerin/Oil/Water, Corr = Correction ring for cover glass thickness (recommend = 0.17 mm), MotCorr = Motorized correction ring for cover glass thickness. There are no dry objectives on these two systems! Always use the correct immersion oil (or demi water for the water immersion lenses).||! There are air and immersion objectives in this system. Do not use oil on the 20x/0.75 Air Objective.
For DLS Illumination there are 1.6x, 2.5x and 5x objectives in the system.
Do not use them for confocal detection !
|Fluo filters:||A, I3, N2.1|
|A for DAPI/Alexa415, I3 for FITC/Alexa488/GFP, N2.1 for Cy3/Alexa568. These filters are used for normal widefield fluorescence mode (for visual inspection) together with the widefield excitation lamp, EL6000.|
|Transmission:||All Oil objectives have a corresponding Differential Interference Contrast (DIC) prism. BF and DIC can be measured with a transmitted light detector in confocal mode.||Same as other microscopes. But for DIC you need another Condensor. So, please use other microscopes if you need DIC.|
|Scanner:||Field of view (FOV) scanner||Tandem Scanner (Field
of view (FOV) and Resonant scanner)
|Field of view (FOV) scanner|
|FOV Scanner: 1800 Hz (3600 Hz bidirectional), Max framesize 8192×8192 pixels, 200° optical field rotation, field diameter 22 mm, 7 fps for 512×512 pixels, 84 fps for 512×16 and zoom 0.75-48x. Resonant Scanner: 8 kHz (16 kHz bidirectional), Max framesize 1024×1024 pixels, 28 fps for 512×512 pixels, 290 fps for 512×16 and zoom 1.25-48x.|
|Detectors:||2x PMT, 2x HyD||3x PMT, 2x HyD(FLIM)||2x PMT, 2x HyD|
|HyD detectors work in photon counting mode. They are at least 2x more sensitive compared to a PMT and have no offset. They have however a limited dynamic range so use a PMT for very bright samples, FRAP Experiments and when you use the Resonant Scanner.|
|Lasers:||405 nm UV Diode 458, 476, 488, 496 en 514 nm VIS Argon 470-670 nm, White Light Laser (WLL) Pulsed, 80 MHz||405 nm UV Diode, 440 nm VIS Diode Pulsed 458, 476, 488, 496 en 514 nm VIS Argon 470-670 nm, White Light Laser (WLL) Pulsed with pulsepicker, 80Mhz||405 nm UV Diode, 470-670 nm White Light Laser (WLL) Pulsed, 80 MHz|
|The WLL gives complete spectral freedom for fluorophore excitation from 470-670 nm. This pulsed laser combined with the HyD detector makes gated detection possible. Gated detection helps removing reflected laser light (from for instance the cover glass) and reduction of autofluorescence.|
|Notch Filters:||445, 488, 514, 458/514, 488/543/633, 488/561/633, 594, 445/594 (in nm)|
|Notch filters can be used to suppress laser reflection|
|Special:||SMD Module for Fluorescence Life-time Imaging Microscopy (FLIM).||Lightsheet Module. 5x, 10x, 25x Widefield Detection with sCMOS Camera. 2.5, 5 water and 7.8 mm Gly twinflect mirrors.|
|Intro:||All microscope can be booked and used after the standard introduction, for standard confocal microscopy. (Please contact us when you use the SP8 DLS for standard confocal use for the first time, we will give a short explanation of the new DMi8 stand and limited, mixed air and oil objectives, Do not use oil on the 20x/0.75 Air Objective.)|
|Extra intro:||Special FLIM/FRET introduction||Special Lightsheet introduction (and help)|