Zeiss LSM880 NLO AiryScan
Use this highly versatile state-of-the-art confocal microscope to solve all your multi-dimensional research questions. Equipped with an NLO module for deep-tissue and label-free imaging, pulsed excitation and time-resolved detection for FLIM, a spectral detector, and an AiryScan detection system for super-resolved fluorescence microscopy, the LSM880 provides a convenient solution for most of your imaging needs.
Use and training
To access this microscope, contact the facility to schedule a hands-on training session. Trained users have 24/7 access to the microscope and are free to schedule experiments using the online booking system. Training can only be given by facility staff.
Acknowledgements
Thank you for acknowledging our imaging facility. You can use the statement below in all your publications that include data acquired on the Zeiss LM880:
We acknowledge the Advanced Optical Microscopy Centre at Hasselt University for support with the microscopy experiments. Microscopy was made possible by the Research Foundation Flanders (FWO, project G0H3716N).
Specifications
Excitation
- HXP120V fluorescence lamp
- CW Argon Laser with 458, 488 and 514nm lines
- CW HeNe 543nm laser
- CW HeNe 594nm laser
- CW HeNe 633nm laser
- Pulsed MaiTai-HPDS laser (690-1040nm)
- Pulsed B&H Diode 488nm laser
- Pulsed PicoQuant Diode 440nm laser
Filter sets
The microscope and scanhead contain a large amount of filter turrets and exchangeable filtercube positions. The following table provides an overview of the standard filter configuration of the LSM880. In addition to these filters, the spectral detection unit (Quasar) allows filter-free selection of the detection wavelength range.
Reflector revolver | MBS invisible light | MBS visible light | SBS emission filter | Airyscan emission filter | Position |
|---|---|---|---|---|---|
FSet43 wf (red) | Plate | MBS 458 | Mirror | Plate | 1 |
FSet49 wf (blue) | None | MBS 458/514 | SBS BP 420-460 + LP500 | BP 420-460 + BP 495-550 | 2 |
FSet38 wf (green) | MBS -405 | MBS 458/543 | SBS LP 525 | BP 420-480 + BP 495-620 | 3 |
BS-MP 355/690+ | MBS -445 | MBS 458/514/594 | SBS LP 570 | BP 420-480 + LP 605 | 4 |
BS-MP-760 | MBS 690+ | MBS 488 | SBS LP 660 | BP 465-505 + LP 525 | 5 |
Rear | MBS 760+ | MBS 488/543 | SBS LP 460 | BP 495-535 + LP 555 | 6 |
MBS -405/760+ | MBS 488/594 | SBS SP 615 | BP 555-620 + LP 645 | 7 | |
MBS -445/750+ | MBS 488/543/633 | Plate | None | 8 | |
MBS 405/488 | MBS T80/R20 | 9 | |||
MBS T80/R20 | Plate | 10 |
Detection
The microscope is equipped with the following detectors:
- Quasar unit Channel 1, S and 2, combined PMT (x2) and GaAsP (x32) detectors.
- Airyscan unit Channel A, 32 channel GaAsP array.
- BiG.2 unit Channel NDD, 2 filter-separated GaAsP detectors.
- T-PMT transmission detector
- optional: B&H HPM-100-40 GaAsP hybrid PMT.
Objective lenses
The microscope is equipped with a variety of low- and high-magnification and NA objectives:
Objective lens | Free working distance | Max resolution * | Immersion | Remarks |
|---|---|---|---|---|
EC Plan-Neofluar 10x/0.30 M27 | 5.2 mm | 1.07 x 11.67 µm | None (air) | |
Plan-ApoChromat 20x/0.80 M27 | 0.55 mm | 0.40 x 1.64 µm | None (air) | |
EC Plan Neofluar 20x/0.50 M27 | 2.0 mm | 0.64 x 4.20 µm | None (air) | |
LD C-ApoChromat 40x/1.10 W Korr UV-VIS-IR M27 | 0.62 mm | 0.29 x 0.87 µm | Water | |
C-ApoChromat 63x/1.20 W Korr M27 | 0.28 mm | 0.27 x 0.73 µm | Water | |
Plan-ApoChromat 63x/1.40 Oil DIC M27 | 0.19 mm | 0.23 x 0.54 µm | Oil | |
Plan-ApoChromat 40x/0.95 Korr M27 | 0.25 mm | 0.34 x 1.16 µm | None (air) |
* Theoretically obtainable lateral (xy) by axial (z) resolution according to the Rayleigh resolution criterium at an emission wavelength of 525 nm.
Lateral resolution (xy) = 0.61 * wavelength / numerical aperture
Axial resolution (z) = 2 * wavelength / (numerical aperture)²
FLIM equipment
- Available wavelengths: 440nm, 488nm and 690-1040nm.
- BiG.2 detector for simultaneous dual channel readout.
- HPM-100-40 Hybrid detector.
- PicoQuant PDL 800-D laser controller and pulse sync driver.
- PicoQuant MultiHarp 150 multichannel Time Correlated Single Photon Counting (TCPSC) device for lifetime recordings during FLIM and FLIM-FRET experiments.
- Dedicated workstation with SymphoTime64, including a 'handshake' with LSM880 control for convenient recording and analysis of lifetime information.
Documentation about Phasor analysis: Find more information here.
Other features
- A PECON incubation system allowing precise control of experimental conditions (Temperature and CO2).
- Zeiss Definite Focus system to counter axial drift during long-term experiments (e.g. timelapse).
Applications
Because the LSM880 is such a versatile setup, it finds use in a large selection of diverse applications, including:
- live deep-tissue two-photon fluorescence microscopy (e.g. migrating cells in live tissue slices).
- labelfree imaging (e.g. SHG imaging of the extracellular matrix).
- fluorescence lifetime imaging (FLIM) (e.g. the visualization of protein-protein interactions using Förster Resonance Energy Transfer (FRET)-FLIM).
- fluorescence correlation spectroscopy (FCS) and raster image correlation spectroscopy (RICS) (e.g. studying membrane receptor mobility and interactions).
- super-resolution airyscan imaging (e.g. precise localization of analytes in stained samples).
- multi-dimensional confocal imaging (xyztc).
- ...