Zeiss Elyra PS.1
Perform fast, live-cell fluorescence imaging experiments on this powerful widefield fluorescence microscope with TIRF and 3D super-resolution (SIM and SMLM) capabilities. This commercial Zeiss microscope is equipped with 4 laser lines (405, 488, 561 & 642nm), an incubation chamber, a dedicated SIM camera and SMLM camera, Definite Focus system and piezo stage. Additionally, the microscope is combined with a patch clamp setup allowing simultaneous fluorescence and electrophysiology recordings.
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 Elyra PS.1:
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
- X-Cite Series 120PC Q fluorescence lamp
- 405nm laser
- 488nm laser
- 561nm laser
- 642nm laser
Filter sets
There are 2 filter turrets with a selection of different filter sets available for use with the Elyra PS.1. Single color experiments are preferentially performed with filter turret 1. Multi-color experiments can be performed with filter turret 2 to allow for fast-frame-switching between different acquisitions.
Filter turret 1 (single band)
position | filter name | excitation | beamsplitter | emission | example fluorophores |
|---|---|---|---|---|---|
1 | Transmission Light | N/A | N/A | N/A | N/A |
2 | FSet77 HE | Triple Band Pass 483 + 564 + 642 (HE) | Triple Beam Splitter 506 + 582 + 659 (HE) | Triple Band Pass 526 + 601 + 688 (HE) | GFP, Alexa Fluor 488, mCherry, Alexa Fluor 568, Cy5, Alexa Fluor 647 |
3 | BP 420-480 + LP 750 | 405 | MBS 405 | BP 420-480 + LP 750 | DAPI, Hoechst, BFP |
4 | BP 495-575 + LP 750 | (405 +) 488 | MBS -488 | BP 495-575 + LP 750 | GFP, Alexa Fluor 488 |
5 | BP 570-650 + LP 750 | (405 + 488 +) 561 | MBS -561 | BP 570-650 + LP 750 | mCherry, mScarlet, Alexa Fluor 568 |
6 | LP 655 | (405 + 488 + 561 +) 642 | MBS -642 | LP 655 | Cy5, Alexa Fluor 647 |
Filter turret 2 (multi band)
position | filter name | excitation | beamsplitter | emission | example fluorophores |
|---|---|---|---|---|---|
1 | Transmission Light | N/A | N/A | N/A | N/A |
2 | empty | N/A | N/A | N/A | N/A |
3 | BP 420-480 + BP 570-640 + LP 740 | 405 + 561 | MBS 405/561 | BP 420-480 + BP 570-640 + LP 740 | DAPI & mCherry |
4 | BP 420-480 + BP 495-550+ LP 650 | 405 + 488 + 642 | MBS 405/488/642 | BP 420-480 + BP 495-550 + LP 650 | DAPI & Alexa Fluor 488 & Alexa Fluor 647 |
5 | LBF -561/642 | (405 + 488 +) 561 + 642 | MBS -561/642 | BP 575-625 + LP 655 | Alexa Fluor 568 & Alexa Fluor 647 |
6 | LBF -488/561 | (405 +) 488 + 561 | MBS -488/561 | BP 500-550 + LP 570 | GFP & mCherry |
Detection
The microscope is equipped with a PCO Edge 4.2 sCMOS camera (6.5µm pixels, 1280x1280) connected to the bottom port, and an Ander iXon+ 897 EMCCD camera (16µm pixels, 512x512) connected to the right-side port.
Objective lenses
The microscope can be equipped with a set of low-magnification air objectives (5x, 10x, 20x and 40x) for fast and easy visualization of samples. Additionally, the microscope is equipped with two high-magnification oil immersion objectives for high-resolution (TIRF, SIM, SMLM) imaging. Additional 1.6x magnification can be achieved using the OptoVar lens (choice between 1x and 1.6x magnification).
- SIM: Plan-Apochromat 63x/1.40 Oil DIC M27.
- TIRF & SMLM: Alpha Plan-Apochromat 100x/1.46 Oil DIC M27. (contains a rotatable TIRF back-reflection blocking pin)
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, super-resolution).
- 3D single-molecule super-resolution imaging with the 3D-PALM slider.
- High precision Piezo stage for fast and precise Z-control.
- Access to a patch-clamp setup for simultaneous fluorescence and electrophysiology experiments.
Applications
Typical research applications of the Elyra PS.1 include:
- patch-clamp fluorometry experiments, combining TIRF microscopy and patch-clamping to study signalling pathways.
- cell-tracking of fluorescently labelled cells passed through cell culture flow chambers.
- super-resolution dSTORM imaging of cellmembrane receptors.
- live-cell super-resolution SIM imaging of calcium signalling.
- single-particle tracking PALM.
- single molecule and image correlation studies.
- ...