Meso-SPIM (L-SPIM)

Unique to the L-SPIM is its geometry: The illumination/excitation objective (NA up to 0.5, air) is used to produce a (~1kHz) digitally-scanned light-sheet in the horizontal plane. The detection objective (8mm working distance, NA 0.95, designed for Scale AS (Scaleview A2 clearing, refractive index n~1.38) - like immersion media) is mounted on a standard inverted microscope frame and images from below (vertically). A correction-collar allows for effective use with media/samples with n=1.33-1.38. If you are using higher refractive index clearing methods, please contact us regarding the possibility of using a different suitable objective lens.

Samples are mounted in/on custom sample holders which are immersed in a small bath. The light-sheet is incident through a quartz plate on the side of the bath, whereas the imaging objective enters the bath from the base through a suitably sealed opening. When used with our custom "Plant Holder" (see below), this configuration allows for clean cross sectional imaging of roots and high resolution 3D mapping of intracellular dynamics for extended periods during growth. On the other hand, using our "Slanted Coverslip Holder", allows for fast 3D volume imaging of cleared organs and small, or the early-developmental-stages of, different model organisms (e.g. C. Elegans, Zebrafish, etc.) that can be mounted directly or indirectly onto the slanted coverslip. The Slanted Coverslip Holder also is also invaluable for routine calibration studies.

Built into the imaging path is a 3-step changeable magnifying optics module which allows for adjustable (optical) magnification of the image on the camera. On the other hand, a motorized iris in the light sheet path allows for adjustment of the effective NA of the light sheet (and thereby the light sheet thickness and on-axis uniformity). By careful adjustment of the imaging and light sheet parameters one can realize an optimal configuration that matches the desired level of detail and field of view for a given study.

The sample holder is in all cases mounted from the top to a long travel range single-axis Piezo (travel range 0.8mm), which is translated to acquire volume images. The piezo stage itself is mounted to 3 DC-motors (travel range 2.5cm) which can also be controlled by a joystick to quickly locate regions of interest.

A custom written user-friendly acquisition software interface (developed in collaboration with Marek Suplata in the group of Katrin Heinze at the University of Wuerzburg), allows for easy control of all imaging parameters and volume acquisition functions, as well as setting arbitrary time-lapse imaging cycles at selectable regions of interest.

Currently the setup is equipped with 4 excitation laser lines (405nm, 491nm, 515nm and 561nm), and images are acquired on a sCMOS camera (Hamamatsu Orca flash 4.0) capable of 100 frames per second.

 

Examples of use:

  • coming soon

 

Microscope Used: Rig1

 

Useful references:

  • coming soon

 

Example from VBCF Advanced Microscopy Facility

  • coming soon

 

Expected Specs:

  • coming soon