Selective Plane Illumination Microscopy – iSPIM & diSPIM

Selective Plane Illumination Microscopy

iSPIM & diSPIM

Selective Plane Illumination Microscopy (SPIM) is an extremely cell-friendly method for imaging live specimens as it only illuminates the region of the sample that is being captured and minimizes phototoxicity and photobleaching.

Applications Include: Cell Dynamics Developmental Biology Live Specimen Imaging Many Illumination Microscopy Applications

Our System’s Advantage…

Low Phototoxicity
>10x Reduction in Photobleaching vs. Confocal

Rapid 3D Imaging
Generate 3D Volumes with Isotropic Resolution (330 nm)

Better Axial Resolution
~2x Better than Confocal

High Acquisition Rates
Up to 200 Images (2-10 Volumes) Per Second

Conventional Sample Mounting
Use Conventional Glass Coverslips or Petri Dishes

Modular and Flexible
Meet Your Application’s Needs

What We Can Provide

ASI offers all of the necessary hardware to implement the diSPIM, which is a flexible and easy-to-use implementation of Selective Plane Illumination Microscopy (SPIM) that allows for dual views (d) of the sample while mounted on an inverted (i) microscope. The diSPIM “head” can be mounted on various inverted microscopes including ASI’s RAMM frame.

ASI manufactures the optomechanical elements, including the motorized stages, 2D galvos for creating and moving the light sheet, and the piezo objective movers. Objectives, lasers, and cameras are required to complete the system; users can procure these other items themselves, use the services of various system integrators selling the diSPIM, or purchase them via ASI.

The diSPIM has been tested successfully on cells cultured on cover slips, cells embedded on collegen gels, c. elegans and zebrafish embryos, and many other samples More information on dispim.org.

Basic SPIM System Configurations

Single-Sided System
Single-Sided Systems (iSPIM)

Light sheet created from one objective and imaged using the other objective. The light sheet is moved through the sample, most often by moving the light sheet using the scanner (galvo) which is synchronized with a piezo stage moving the imaging objective.

Advantages: Fastest acquisition, least expensive, straight-forward set-up.

Disadvantages: Better XY resolution than Z resolution.

Double-Sided System
Double-Sided Systems (diSPIM)

Both sides have a light sheet scanner, piezo objective positioner, and camera. During an experiment a stack of images is collected from both views, and the two datasets can be merged computationally to yield a 3D dataset with isotropic resolution (the usual problem of poor axial resolution is overcome by information from the other view). Can operate in single-sided mode if desired.

Advantages: XY & Z resolutions are all very good – yielding a combination of speed and resolution that is unsurpassed for live cell imaging.

Disadvantages: More hardware to buy. Data post-processing required for isotropic resolution.

Variations
Varitions in SPIM Systems
  • Photomanipulation using inverted microscope
  • Filter wheels on imaging paths
  • Asymmetric single-sided system, e.g. using same objectives as Lattice Light Sheet
  • Non-gaussian beam
  • 2-photon microscopy

More Info

Videos & More

SPIM Videos

Published Papers on the iSPIM/diSPIM System

Webinars

New Technologies for High Spatial and Temporal Resolution Imaging