1. TIRF MICROSCOPY
TIRF Labs offers a family of TIRF Microscopy systems with different geometries:
prism-based, lightguide-based, and objective-based TIRFM. Each geometry has its own
set of advantages and limitations. For example, p-TIRFM provides the best signal-to
background ratio and is well-suited for in vitro studies with closed flow cell surrounding
the TIRF area, but is too difficult to implement p-TIRFM with open perfusion chamber on
an inverted microscope. lg-TIRFM and o-TIRFM are better-suited for live cell studies with
open perfusion chambers. Both p- and lg-TIRFM are factory-aligned systems with highly
reproducible intensity of the evanescent wave; they are well-suited for multicolor TIRF
experiments (see Application Notes at www.tirf-labs.com). o-TIRFM requires alignment,
experiences irreproducible intensity of the evanescent wave, but allows to vary the angle
of incidence, and hence, the depth of penetration. All three TIRFM geometries efficiently
reject background fluorescence and scatter and are capable of super sensitive detection,
down to single molecules. Selecting of geometry and particular configuration of TIRF
system depends on your application. TIRF Labs will help you in this process. Please read
the brochure “Compare TIRF Geometries” and TIRFM application notes, and email or call
us for our recommendation for TIRF geometry and specific configuration
1.1. p-TIRFM - prism-based Total Internal Reflection Fluorescence Microscopy. p-TIRFM is
a geometry, which provides the best signal-to-background ratio. TIRF Labs offers p-TIRFM flow
system with closed flow cell and advanced fluidics, which is well-suited for handling small liquid
aliquots, down to sub-microliter volumes. p-TIRFM can be used
with dry, water-, and oil-immersion objectives. It uses silica
prism and silica or glass slides. p-TIRFM is a factory aligned
system supplied as an add-on accessory for rapid installation on
inverted microscopes. p-TIRFM fits into 128 mm x 85 mm
window that holds 96-well SBS plate. It can be used with Ludl
and Prior motorized stages, as well as manual XY-translation
stages. Base model is supplied with one of the illuminators: 405 nm, 450 nm, 532 nm, or 630 nm
installed at the p-TIRFM platform. Multicolor illuminators are available as options.
1.2.1 lg-TIRFM-170 - lightguide-based Total Internal Reflection Fluorescence Microscopy
is a geometry, which exhibits excellent signal-to-background ratio, close to that of p-TIRFM
geometry. Similar to p-TIRFM, the excitation light does not travel through the emission channel,
providing to lg-TIRFM signal-to-background ratio an order of magnitude better than that for o-
TIRFM. lg-TIRFM is a factory-aligned system with fixed angles and reproducible intensity of the
evanescent wave, which is well-suited for multicolor TIRFM experiments. lg-TIRFM can be used