Leica Stellaris 8 DIVE upright Confocal, multi-photon
This is a combined multi-photon and confocal, upright-configuration microscope that is designed for deep, multi-parametric in vivo and ex vivo tissue and cell imaging using both 1-photon and 2-photon excitation for simultaneous and sequential imaging of a wide variety of expressed fluorescent proteins and endogenous signals. The DIVE microscope incorporates Leica’s 4Tune non-descanned detector head with 2 freely tunable-filter channels with high sensitivity hybrid detectors. This enables 2 channel, spectral separation of up to 2 fluorescent signals simultaneously and an unlimited number of signals sequentially. The Stellaris 8 microscope is also configured for confocal imaging with six solid state lasers. The Stellaris 8 DIVE can be used interchangeably in 1-photon (confocal) or 2-photon excitation imaging modes. It is fully enclosed for heating and environmental control with media perfusion capabilities and CO2 control.
The Stellaris 8 DIVE is incompatible with multiwell plates or chambered coverslips due to the upright configuration that requires 'dipping' the lens into the media for live cell imaging (see inverted setups such as Leica SP8, Zeiss LSM880 for those needs).
System specifications
- STELLARIS 8 with Focus on DIVE and CRS
- DIVE
- FALCON prepared for WLL or DIVE
- DM6 CFS
- 2 pos + single
- BF Detector for DM 6 and DIC
- LED3
- Filtersystem BGR ET, size k
- Maerzhaeuser Motor Stage
- Super Z Galvo Stage
- Insert Universal Type H
- RGTBV
- Notch Filter Set VIS base
- Laser 405 DMOD
- 8 kHz Tandem Scanner DIVE
- Power HyD S, Position 1
- Power HyD S, Position 2
- Power HyD X, Position 3
- Power HyD S, Position 4
- CUDA Workstation Expert
- Single Beamrouting DIVE
- AOM
- 4Tune 2 channel spectral detection
- 4Tune HyD Position 1
- 4Tune HyD Position 2
- TLD 2 channel for DM6 CFS Dichroic
Objectives
|
Magnification |
N. A. |
Resolution x,y/z (nm) |
Working Distance (um) |
|
HC PL Fluotar 10X, air |
0.30 |
650, 4770 |
11000 |
|
HC PL APO 20x/IMM CORR CS, H2O/Glycerol/oil |
0.70 |
280, 1290 |
250 |
|
HCX PL APO 40X, oil |
1.25-0.75 |
160, 335 |
100 |
|
HCX PL APO 63x/1.30 GLYC CORR CS 21 C |
1.3 |
160, 335 |
280 |
|
optimal for multiphoton: |
- | - | - |
|
HCX APO L20x/1.00 water imm |
1.0 |
140, 236 |
2000 |
3 other specialized “dipping” objectives with long working distances >1 mm for imaging optically cleared samples:
1. HC FLUOTAR L 16x/0.6 IMM CORR VISIR, WD 2.5 mm (M32). This FLUOTAR lens is optimized for immersion media with refractive indices 1.33 – 1.52 and will work for iDISCO cleared samples.
2. HC FLUOTAR L 16x/0.8 IMM motCORR VISIR, WD 8.5 mm (M25). This FLUOTAR lens is an extremely long working distance lens for deep, cleared-tissue imaging and it is optimized for immersion media with refractive indices 1.33 – 1.46. It has the smaller thread for mounting on the 2-position nosepiece lens holder.
3. HC FLUOTAR L 25x/1.00 IMM motCORR VISIR, WD 6.00mm (M32). This FLUOTAR lens is optimized for immersion media with n=1.457, dedicated to CLARITY cleared samples. Number 2 and 3 FLUOTAR lenses are equipped with a motorized correction collar required for fast and easy adjustments, without bumping the sample out of focus, to optimally correct for changes in refractive index. All three objective lenses are optimized for high transmittance in the visible and infrared (IR) spectrum (typically 470 -1200 nm).
Lasers
1p Lasers lines: 638nm, 561nm, 514nm, 488nm, 448nm, 405nm, 730nm
2P Laser: Spectra Physics 15W MaiTai DeepSee Tuning range 690-1040 nm
Variable Scanner 1 - 1400 Hz Line frequency max: 2800 Hz 5 Images/second, 512 x 512 ~54 Images/second, 512 x 16 Max. Image resolution 8192 x 8192 Line frequency up to 2800 Lines/second.
Detection
4 channel descanned/confocal detection: 4 descanned Hybrid-GaAsP detectors for increased light sensitivity, enables advanced use such TAU Separation.
4 channel non-descannced detection (NDD): 2 reflected, 2 transmitted/SHG for multi-photon imaging
1 transmitted light detector
FLIM
Leica Tau excitation DIVE - TauSense (we are seeking funding for full FLIM functionality provided by FALCON option.)
See CSIF staff for more information on FLIM imaging.
Off-hour credits
Labs that use the following microscopes: Zeiss LSM880, Leica Stellaris8, and/or Leica SP8 for 25 or more hours per month, during OFF-HOURS will be rebated to reach $32/hour for all use of that microscope.
Labs that use the Zeiss LSM880, Leica Stellaris8, and/or Leica SP8, for 75 or more hours per month, during OFF-HOURS will be rebated to reach $23/hour for usage. This rate is in recognition of the high number of hours required for continuous overnight and weekend imaging inherent in the use of these microscopes for live cell imaging.
To qualify for these rates you must have after-hours access to the facility (i.e. be an experienced, competent user) and must use the confocal/2 photon between 5:00 pm and 8:00 am M-F, holidays, or any time on weekends. Reserved but unused time does not qualify for these discounts. Please speak with the facility Director Jon Mulholland for more information.
Funding and Required Acknowledgement
This microscope was funded via a NIH S10 SIG to Dr. Lucy O'Brien. For all data published using this microscope you must acknowledge NIH ORIP funding: 1 S10 OD032300-01