Lightsheet Microscopes
The SCF houses a dual-side illumination lightsheet microscope: the Z1 from Zeiss. This selected plane illumination microscope (SPIM) achieves optical sectioning with a thin light sheet. This technology illuminates only the sample in the observation plane and has therefore a low phototoxicity/bleaching. Having no need for pinholes as used in confocal microscopy results also in no loss of excitation light. The signal intensity allows for short exposure times and therefore for a high temporal resolution.
This microscope enables users to acquire both live and fixed (cleared) samples, in a wide range of sizes and in different numbers.
For detailed descriptions of the individual workstations (all specifications, filter settings, etc.) and interactive spectra viewers, see our internal Wiki (D-BSSE login required).
Zeiss Z1 Lightsheet (Zeiss Z1)
The Z1 lightsheet microscope is a versatile microscope, capable of imaging both live and fixed / cleared samples. The system is designed to mount samples vertically, using a glass capillary and hydrogel solution, and to suspend it in medium / clearing buffer. The dual-side illumination, the pivot scan that eliminates the shadows generated by any “obstacle” in the light path, and the multi-view acquisition allow to reliably obtain very uniform images. The dual camera combination enables the acquisition two fluorescence channels simultaneously, speeding up multicolor acquisitions.
The system is equipped with 2 powerful workstations, one dedicated to the acquisition control with ZEN Black software, and another dedicated to data collection and analysis. This configuration allows to run experiments while data are transfered to central storage or analysed without conflicts.
Setup
- Dual illumination setup (allows better light penetration into the sample)
- Dual camera detection module: 2x PCO Edge sCMOS
- Multi-position imaging (XYZ + rotation)
- Immersion sample chamber for: aqueous media (n=1.33) or clearing media (n=1.38, n=1.45)
- Temperature-, CO2-, and humidity-controlled
Excitation
- Laser lines: 405 nm, 445 nm, 488 nm, 515 nm, 561 nm, 638 nm
- Illumination optics: 5x/0.1, 10x/0.2
- Filter combination: 420-470nm + 525-545nm; 420-470nm + 575-615nm; 505-545nm + 575-615nm; 505-545nm + LP 585nm;
- Light sheet thickness: 2um - approx. 14 um
Detection
- Detection optics for live acquisitions: 5x/0.16 Dry, 10x/0.5 W, 20x/1.0 W, 40x/1.0 W
- Detection optics for cleared tissue acquisitions: 5x/0.16 Dry, 20x/1.0 Clearing (n 1.43-1.48)
- Pixel size 6.5 um
- Max image resolution 1920 x 1920 pixels
- Bit depth: 16 bits
- Max frame rate: 30 fps at 1000 x 1000 pixels
