Total Internal Reflection Fluorescence

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Single Molecule Detection, Super Resolution Microscopy, sm-FRET, Single Cell Analysis,  Assay Development, Real-time Microarrays, Cell Membrane Studies,

TIRF microscopy, TIRF accessories for fluorometers, TIRF electrochemistry, TIRF dielectrophoresis, iDiagnostics - cellphone based TIRF, and more>>>

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TIRF Microscopy  

TIRF for Fluorometers


TIRF Biosensors

TIRF Slides, TIRF Prisms

Total Internal Reflection Fluorescence (TIRF)

Single Molecule Detection, Super Resolution Microscopy, Real-time Microarrays,

Plasma Membrane Studies, Single Cell Analysis, iDiagnostics, and more >>>

  TIRF Principles. TIRF employs the phenomena of total internal reflection and the evanescent wave to selectively excite fluorophores located at the surface and in close proximity (~100 nm) to the surface of a glass, silica, or plastic slide. Fluorophores that are in the bulk of solution are not excited and do not fluoresce. Exceptional spatial selectivity enables TIRF with numerous advantages, including the ultimate limit of detection - single molecule. TIRF effect allows for circumventing the diffraction limit of  resolution and, therefore, is used in many super resolution microscopy methods.  TIRF efficiently minimizes the background and allows for the real-time monitoring  the molecular dynamics at the level of single molecules.  More »

    TIRF Applications. TIRF spectroscopy and TIRF microscopy are powerful analytical tools for numerous life science areas, including single molecule detection, super resolution microscopies, cell biology,  real-time microarrays for molecular diagnostics, bioassay development, nanoengineering, and drug screening. In particular, TIRF is “a method uniquely suited to image the plasma membrane with its associated organelles and macromolecules in living cells. The method shows even the smallest vesicles made by cells, and can image the dynamics of single protein molecules.” [Steyer JA, Almers W. A real-time view of life within 100 nm of the plasma membrane. Nature Reviews, Molecular Cell Biology, 2001, 2(4), 268-75]. References to thearticles published by our customers and TIRF application notes are available at www.tirf-labs.com/applications.html   More »

    TIRF Products. TIRF Labs offers a broad range of TIRF products: prism-, lightguide-, and objective-based TIRF microscopy systems,  from simple add-on accessories - to sophisticated computer-controlled  turnkey stations. Our LED and laser fluorescence illuminators include cost-efficient single-color light sources, two-, three-, four-color,  and seven-color computer-controlled  illuminators. For TIRF spectroscopy we supply TIRF accessories with gravity flow and digital fluidics. Our automated TIRF microarray biosensors detect multiple nucleic acid, protein, and metabolite markers in parallel and in a matter of a few minutes. Popular products also include TIRF slides, lightguides, and prisms. More »

Technologies Applications Applications Applications

Select TIRF Geometry

Digital Fluidics SmartFlow

iDiagnostics - The Cellphone Future of Precision Medicine  visit: i-Diagnostics.net

TIRF Products  

 uTIRF Brochure

   TIRF Geometries.  Prism-, lightguide-, and objective-based optical schemes are three popular geometries that are used for TIRF microscopy. Each geometry has its own set of advantages and limitations. A Prism-based scheme provides the best signal-to-background ratio, but is difficult to implement with an open perfusion chamber on an inverted microscope. A lightguide-based geometry yields exceptional flexibility and superior signal-to-noise ratio, can be used with open perfusion chamber on inverted microscopes, is compatible with dry, water-, and oil-immersion objectives, but requires larger optical power to obtain equal intensity of the evanescent wave. An objective-based scheme collects the largest portion of the emitted fluorescence, but the quality of TIRF effect is severely compromised, because the excitation light shares the same optics with the emission channel; large intensity (~15%) of the stray light excites the bulk of the specimen and deteriorates the TIRF effect produced by objective-TIRF. TIRF Microscopy webpages will help you to choose a geometry suited for your TIRF applications. TIRF Products webpages will help to select illuminators, fluidics, optional electrochemical and dielectrophoretic control, other devices and supplies that are necessary  or useful for TIRFing. TIRF Labs offers the entire range of TIRF instrumentation: from inexpensive add-on TIRF accessories for microscopes and fluorometers -to turnkey TIRF microscopy stations and automated TIRF biosensor instruments.  More »

Prism-TIRF Microscopy  

Lightguide-TIRF Microscopy

Objective-TIRF Microscopy

Visit also:   www.TIRFmicroscopy.com

TIRF microscopy for single molecule detection and super resolution methods

In addition to traditional objective-based TIRF, TIRF Labs is offering advanced prism- and lightguide- based TIRF systems that feature superior optical schemes. In prism- and lightguide-TIRF schemes the excitation path is separated from the emission channel. Unlike in objective-type TIRF, the intensity of stray light in lg- and p-TIRF is minimal; the TIRF effect is clean. Lg-TIRF and p-TIRF are exceptionally flexible geometries - can be used with low and high numerical aperture lenses, dry, water-, and oil-immersion objectives, UV, visible and near-IR excitation.


Use the uTIRF turnkey station,

or take prism-, lightguide-, and objective-TIRF

as accessories for your microscope

Introducing uTIRF Cube

Ultimate turnkey TIRF microscopy station

uTIRF Cube consists of

prism-, lightguide-, and objective-based TIRF;

4X, 20X, 40X, 60X, or 100X objectives;

low light CMOS camera;

and 465/532/638 nm illuminator

Main Advantages / Limitations

- p-TIRF: The cleanest TIRF effect /

 Closed flow cell - no open perfusion

- lg-TIRF: Clean TIRF effect /

 Needs to align TIRF area with objective

- o-TIRF: TIRF area travels with objective /  

Evanescent wave contaminated (~15%) by stray light

  uTIRF Cube -  ultimateTIRF microscopy turnkey station


uTIRF Cube - single molecule detection guaranteed


For Application Notes and published papers click here

To compare TIRF microscopy geometries click here

To minimize stray light in TIRF microscopy click here

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