Inverted Zeiss LSM 780 multiphoton laser scanning confocal microscope
The microscope is coupled with a Spectra Physics MaiTai, DeepSee ultrafast pulsed laser system for multi-photon excitation deep within tissue. It is equipped with the LSM BiGmodule which provides two highly sensitive GaAsP detection channels (45% QE compared to ~25% QE for conventional PMT). It is fully enclosed for heating and environmental control with media perfusion capabilities and CO2 control. Additionally, the microscope has a single channel fluorescence lifetime channel for FLIM measurements.
Applications
- Deep tissue imaging
- Imaging of weak fluorescence signals
- Live Cell Imaging
- Spectral Imaging, Unmixing
- Excitation Fingerprinting
- Super Resolution imagining with AirScan (1.7X improvement over diffraction limited imaging)
- Ion Imaging
- FLIM
- FRET and Anisotropy
- FRAP and FLIP
- FCS
- Photoactivation/ -conversion
- Photon Counting
- FLIM with Becker Hinkl SPC-150, detector : HPM-100-40
System specifications
- Inverted Zeiss Axio Observer Z1 microscope with Definite Focus
- Motorized XY Stage, allowing large images and multi-point time-lapse imaging
- A single (34-spectral-detection-channels) GaAsP photodetector plus two PMTs
- BiG Module with 2 GaAsP photodetector
- MaiTai DeepSee?(allow highest pick power)
- 2-photon microscopy with non-descanned BiG detectors
- Spectral unmixing, spectral fingerprinting and photon counting
- InVivo Scientific enclosure w/ environmental control (temperature and CO2)
- Up to 8 frames/sec with 512 x 512 pixels (max. 250 frames/sec with 512 x 16)
- Up to 6144 x 6144 pixels resolution
- Detection from 390 to 750 nm freely configurable
Software
- Zen Black
- Zen Blue
Zen lite free download page here:
http://www.zeiss.com/microscopy/en_us/products/microscope-software/zen-lite.html#downloads
Objectives
|
Name |
Magnification |
NA |
Immersion |
working distance |
|
EC PLAN NEO |
10x |
0.3 |
air |
5.2mm |
|
PLAN APO |
20x |
0.8 |
air |
0.55 mm |
|
EC PLAN NEO |
40x |
1.3 |
oil |
0.21 mm |
|
C-APO |
40x |
1.2 |
water |
0.28 mm |
|
PLAN APO |
63x |
1.4 |
oil |
0.19 mm |
|
LCI PLAN NEO |
25x |
0.8 |
Oil, Gly, Water |
0.57 mm |
Laser lines
1P lasers:
- 405nm
- 458nm
- 488nm
- 514nm
- 561nm
- 594nm
- 633nm
Laser Power measure out of the 20X Dry (in nW) - 05/08/2019
| Laser Wavelength in nm | 1% | 10% | 100% |
|---|---|---|---|
| 405 | 41 | 194 | 1850 |
| 458 | 494 | 4490 | 49200 |
| 488 | 50 | 337 | 3260 |
| 514 | 37 | 227 | 2170 |
| 561 | 208 | 1930 | 18300 |
| 594 | 15 | 32 | 194 |
| 633 | 24 | 151 | 1360 |
2P laser
- Tuning range: 690-1040nm
Detection
1x 32-anode Hybrid-GaAsP detector for increased light sensitivity/spectra unmixing, 2 standard fluorescent photomultiplier tubes (PMT).
FLIM
The pulsed ultra-fast MaiTai laser allows the LSM780 confocal to be used for fluorescence lifetime measures. FLIM measurements are done using the Becker Hinkl Inc., Simple-Tau 150 system (http://www.beckerhickl.com/simpletau152.htm) with a single ultra-fast Time Correlated Single Photon Counting TCSPC channel. These electronics measure the time between the laser pulse and the arrival of photons at the detector; this constitutes the basis of the FLIM measurement. FLIM is the best way to do FRET measurements as only the donor lifetimes need to be measured. (NOTE: use of the FLIM hardware requires coordination with CSIF staff.)
Off-hour credits
- Labs that use the following microscopes: Zeiss LSM780 or Nikon-SDC for 25 or more hours per month, during OFF-HOURS will be charged $30/hour for all use of that microscope.
- Labs that use the Zeiss LSM780 or Nikon SDC, live cell image system 75 or more hours per month, during OFF-HOURS will be charged at $20/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, or any time on weekends. Reserved but unused time does not qualify for these discounts. Please speak with the facility Director for more information.
Funding
This microscope was funded by jointly by the SOE and the Beckman Center (Dr. Alex Dunn and Jon Mulholland, PIs). Installation date: 09/2014