Super-resolution Microscopes: Roper iLas 2 Microscope (Level 5 Imaging Suite)

Inverted /Single molecule imaging (PALM / STORM) microscope.

The Roper iLas 2 instrument provides a full 360º TIRF illumination for an even plane of excitation ideal for single molecule imaging and particle tracking in live cells.

For more information on these techniques and to request training please use the PPMS booking system.

Features Include:

Fully motorised X-Y-Z stage
Evolve Delta EMCCD for rapid Single Molecule Imaging (62.5 fps @ 512×512 20MHz)
Piezo Z-drive and Nikon perfect focus system
iLas2 doube laser illumination – http://www.roperscientific.fr/brochures/iLas.pdf
Azimuthal spinning averages TIRF polarization effects and makes a complete TIRF image
No interference fringes and shadows
Same penetration depth for different wavelengths in time sharing mode
Wiefield or oblique illumination using angles below the critical TIRF angle

Objectives:

Air: 10x (0.45 NA)
Oil: 100x 1.49 NA for single molecule imaging

Fluorescent Filter Sets:

Dual Green-Red filter (allows viewing of 488 and 560 emitting fluorophores together)
QUAD filter (simultaneous blue, green, red and far-red)

Lasers:

405nm 100mW Vortran
491nm 150mW Cobolt Calypso
561nm 150mW Cobolt Jive
642nm 100mW Vortran

Laser Power:

Measurement at BFP with 100x

405nm (3.61 mW),
491nm (4.1 mW),
561nm (19.9 mW),
642nm (9.8 mW)

The Roper iLas2 sptPALM has the Panlab LE12106 electrical stimulation system, which provides a wide range of stimulations with single or repetitive pulses of selectable constant voltage or current. For more information on the Panlab LE12106, please go to the instrument page at:
https://support.harvardapparatus.com/hc/en-us/articles/4402627586451-PANLAB-LE12106-Single-Channel-Digital-Stimulator

The Roper also has the Warner Instruments VCS-6 perfusion system, a computer-controlled valve control system that is a multi-valve perfusion setup designed to automate and control the delivery of solutions to imaging and recording chambers. For more information on the Warner Instruments VCS-6, please visit the instrument page:
https://www.warneronline.com/VCS-computer-controlled-Valve-Control-System

For these experiments, we use the Warner Instruments multi-well insert QE-1 quick exchange platform and RC-37FS field stimulation chamber on the Okolab stage-top incubation chamber:

https://www.warneronline.com/quick-exchange-platform-for-35-40-mm-dishes-qe-1

https://www.warneronline.com/sites/default/files/2018-09/RC-37WS%2037FS%20%20%282004.02.05%29.pdf

The Roper iLas2 sptPALM with the Panlab LE12106 electrical stimulation system and Warner Instruments VCS-6 perfusion system

User guide here.