The Intelligent Imaging Innovations (3i) lattice light sheet microscope (LLSM) is state-of-the-art for rapid high-resolution live cell imaging with low phototoxicity stemming from efficient lightsheet excitation/collection. Compared to a spinning disk confocal, the LLSM has 100x less photobleaching in dithered mode and also provides better axial resolution.
- An ultra-thin lightsheet achieves optical sectioning for high resolution 3D imaging over time (100s of images per second).
- Cell viability is maintained by only illuminating what can be captured--resulting in low peak light intensity and lowest total light dose.
- 5% CO2 and temperature control (with active feedback) for imaging at physiological conditions
- A motorized annular mask enables us to test and match lightsheets to your sample on-the-fly.
Three modalities to match spatiotemporal requirements
- Dithered mode (standard): fastest acquisition speeds with least photodamage, one image per plane
- SIM (structured illumination microscopy): improves xz resolution, multiple images per plane
- SRRF (super-resolution radial fluctuations): improves xy resolution, many images per plane
Objectives
Excitation objective: 0.7NA water objective, 3.7mm WD
Detection objective: Nikon 25x/1.1NA, 2.0mm WD, correction collar, 62.5x total magnification
Laser lines (nm)
- Typical sample: cultured cells or samples mounted onto a 5mm glass coverslip
- Sample and objectives will be submerged in a 2.5mL or 12mL media bath.
- Careful selection of fluorophores necessary to prevent cross-excitation
- 30 minutes between sample loading and start of acquisition for thermal equilibrium
- Field of view: The stage is limited to 100um of lateral movement during acquisitions
- Accessible regions of the coverslip are roughly a 2x2mm box in the middle of the coverslip
- No adaptive optics, but we're happy to test thicker samples.
- Tiling not possible.
- Better resolution (at the cost of light dosage) is obtainable through our 3D-SIM microscopes
- Low throughput, but the low light dosage makes some live cell experiments possible
- You will generate 100s of GBs to TBs of data and must have a data storage plan.
Primary contact
Stoyan Ivanov stoyan.ivanov@emory.edu