Localize Single Molecules with Unrivalled Precision
ELYRA P.1 takes light microscopy to the limit. By localizing small structures and even single molecules, you are able to achieve resolutions of down to 20 nm laterally and 50 nm axially.
You are interested in processes that take place near the coverslip. You want to see and measure single molecules in or near the plasma membrane like lipid rafts, receptor clustering or cell-substrate adhesion sites. With 3D-PALM you use photo-switchable proteins and profit from an excellent z capture range.
The patented exclusive PALM technology of ELYRA P.1 takes you into a new world of data quality. Detection with an effective resolution down to 20 nm will show you substructure and patterns where conventional light microscopy will simply show co-localization.
As a single molecule method, PALM is inherently quantitative – every image is a molecular statistics experiment.
•Illumination module Fully motorized Epifluorescence (EPI), high inclined and laminated optical sheet (HILO) and total internal reflection illumination (TIRF);
simultaneous TIRF illumination with VIS and 405 nm laser lines; individual triggering of lasers for synchronizing dye activation and illumination to camera read-out and transfer times; motorized TIRF angle adjustment; motorized TIRF field adjustment with three field size options.
•3D-PALM module Double phase ramp in pupil plane of back aperture of objective providing for phase ramp imaging localization microscopy (PRILM);
z capture range typically 1.4 μm; multi-plane acquisition possible to extend z range
•Cameras EMCCD camera (mounted to side port of microscope; left side port without LSM, right side port with LSM)
•Objective lenses (PALM) alpha “Plan-APOCHROMAT” 100x/1,46 Oil DIC, alpha “Plan-APOCHROMAT” 100x/1.57 Oil-HI DIC Corr (2D-PALM);
alpha "Plan-APOCHROMAT" 63x/1.4 Oil DIC, C-APOCHROMAT 63x / 1.2 W Corr DIC (3D-PALM)
•Imaging modes “Widefield (WF)” mode (sample illumination with arc lamp), "Laser WF" mode (sample illumination with laser),
“LSM” mode (available if combined with LSM 710 or LSM 780)
•Field of view (PALM) Maximal field of view 51.1 x 51.1 μm (with alpha “Plan-APOCHROMAT” 100x/1,46 Oil DIC, full chip recording);
81.1 x 81.1 μm (with "Plan-Apochromat" 63x/1.4 OIL DIC, full chip recording); HP field 2 x smaller, uHP field 2 x √2 smaller than TIRF field
•Localization precision (PALM) Typically 20 nm – 30 nm lateral, 50 nm – 80 nm axial, given sufficient signal-to-noise
•Multi-color imaging (PALM) Detection of up to two different fluorescent labels (sequentially or quasi simultaneously by fast sequential laser switching)
•Acquisition speed (PALM) TIRF (PALM) and widefield mode: up to 30 frames per second (full frame mode, 512 x 512 pixels);
>100 frames per second in sub-array mode
•Data recording and analysis (PALM) Full software control of PALM imaging; software holding focus based on fiducial markers
Online PALM processing for simultaneous data acquisition and analysis (2D-PALM only);
manual editing of parameter settings for optimal results in PALM with different fluorophores;
feature-rich rendering of PALM localization tables; export and import of localization tables for custom filtering;
correction algorithms for lateral and axial drift;
chromatic aberration correction (based on fiducial markers or prominent structures)
Multi-emitter fitting algorithms allow to analyze overlapping signals with high precision.
Up to 10 times higher labeling densities are possible speeding up acquisitions by the same factor.