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.

Picture of a Zeiss AxioObserver microscope stand, equipped with a dark PECON incubator and an Elyra PS.1 superresolution imaging module. Picture of a Zeiss AxioObserver microscope stand, equipped with a dark PECON incubator and an Elyra PS.1 superresolution imaging module.

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.
  • ...

Performance

Laserpower stability

Recorded laserpower when requesting 1% transmission. Laserpowers are recorded on the microscope stage through a 10x/0.3 objective lens.

[include laserpower graphs]

Laserpower linearity

Recorded laserpower at various requested % transmission. Laserpowers are recorded on the microscope stage through a 10x objective lens. Results of the most recent laserpower measurements are shown in a log-log plot.

[include laserpower linearity graphs]

Contact

Sam Duwé - AOMC

Location
Agoralaan, Building C, 3590 Diepenbeek
Function
Microscopy expert