Zeiss LSM880 Fast AiryScan, room 3103

Information about the microscope

Point-scanning confocal set up for live imaging (heated, humidified incubation chamber, CO2 control 0-50%). Fully motorized xy stage for multi-position and tile scan, z Piezo for fast 3D stacks. The system is equipped with the Definite Focus.2 (hardware autofocus).

Water-cooled scanning unit for higher scan speed. Confocal imaging using 3 detectors, one of which is a more sensisitve GaAsP detector. This detector can also be used for simultaneous spectral imaging of 32 channels at 9 nm spectral resolution covering the visible range (410-695 nm). This is ideal for ratiometric measurements, e.g. for FRET, biosensors, and for the separation of highly overlapping fluorescent dyes via linear unmixing (up to 8 color channels simultaneously). The flanking detectors are optimized for UV-vis (370 nm and above) and far-red (up to 760 nm), respectively.

Super resolution imaging using AiryScan: additionally the microscope is equipped with the AiryScan detector, which has all the capabilities (rejection of out-of-focus light, 3D imaging) of a confocal, without using a pinhole. Instead, the emission point-spread function (1.25 Airy units) is projected onto a 32-element GaAsP array detector. This enables imaging at higher signal to noise ratio. The individual detector elements act as virtual pinholes of very small diameter (0.2 AU), which together with the increased SNR and linear deconvolution increases the resolution almost 2-fold compared to conventional confocal (120 nm lateral and 350 nm axial resolution). No special fluorophores or sample preparation are needed.

The Fast module for AiryScan shapes the excitation spot into an ellipse, the AiryScan array detector is then used to detect 4 pixels simultaneously, increasing scan speed 4-fold (27 fps for 480x480) while still improving resolution significantly compared to conventional confocal (170 nm lateral).

For more information on AiryScan see the Zeiss website: LSM880 AiryScan

Laser lines

* from fiber

Objectives

* from coverslip #1.5 (170 µm)

** effective field of view at 1x confocal zoom