Lightsheet - MesoSPIM (Irchel)

MesoSPIM

The mesoSPIM (mesoscale selective plane illumination microscopy) is the ideal instrument to quickly bridge scales from the µm- to the cm-level, which enables it to serve as an excellent tool for detailed three-dimensional anatomical investigations in many biological and medical fields (Voigt et al., 2019).

Main features:

  • Compatibility with all major clearing techniques Clarity and iDISCO.
  • Large field-of-views, ranging from 2 to 20 mm using an Olympus MVX-10 macroscope.
  • Dual-sided excitation. Optimized for fast screening of whole mouse brain samples, yielding relatively small datasets (10-14 GB/brain/color) near-isotropic sampling (5-10 µm in X,Y & Z) due to an axially swept lightsheet which leads to uniform z-resolution across the field-of-view.
  • Modular sample holders and quick sample exchange.
  • Large travel range (45 x 45 x 100 mm) to allow imaging of very large samples.

For further information: MesoSPIM.org

Location

University Zurich, Irchel Campus, Room Y44-J-33.

Training Request

Follow this link to apply for an introduction to the microscope.

Technical Specifications

Excitation path

The mesoSPIM excitation path is based on axially scanned light-sheet microscopy (ASLM). The waist of the light-sheet is translated through the sample in synchrony with the rolling shutter of the camera. The waist is translated with electrically tunable lenses by Optotune.

Light Sources and Lasers

The iChrome MLE comprises 3 diode lasers and one DPSS laser fully integrated in one compact housing. Lasers: 405nm (100mW), 448nm (100mW), 561nm (100mW), 640nm (100mW)

Detection path

The mesoSPIM detection path consists of an Olympus MVX-10 macroscope with a MVPLAPO 1x objective, which allows FOVs of 2-20 mm in combination with a Hamamatsu Orca Flash 4.0 V3 camera.

Fluorescence Filters

Name   Emission filter  
Quadband filter 405/488/561/640   BP444/27; BP523/22; BP594/20; BP704/46  
508   BP 520/35  
515 LP   LP 515  
561 LP   LP 568  
594 LP   LP610  

Camera Systems

A Digital sCMOs-Camera: Pixel Array 2048 x 2048 (pixel size 6.5 µM), (Hamamatsu ORCA-FLASH 4.0 V3).

Software

The microscope software, mesoSPIM-control, is written in Python and allows users to specify sequences of z-stacks using a table-based acquisition manager. The software can also be used to acquire large-scale tiling acquisitions.

Accessories

Remarks

  • For initial tests and the first experiments, the ZMB will serve users with the needed experimental accessories such as cuvettes and immersion solutions. If they become regular user of the microscope, they have to organize the accessories by themselves.

Literature and Links

Further information (internal UZH use only)

Follow this link for further background information, documents and links.

Responsible Persons

If you have questions about the microscope please contact José María Mateos or Dominik Haenni.