Lambert Instruments

Typically, the LIFA -Lambert Instruments Fluorescence Attachment is a widefield FLIM -fluorescence lifetime imaging microscopy system with a modulated LED light source. However, other modulated light sources like diode lasers can also be used to perform FLIM -fluorescence lifetime imaging microscopy. This makes the LIFA -Lambert Instruments Fluorescence Attachment suitable not only for widefield lifetime images, but also for confocal lifetime images. Confocal microscopy is an imaging technique used to increase micrograph contrast and/or to reconstruct three-dimensional images by using a spatial pinhole to eliminate out-of-focus light in specimens that are thicker than the focal plane.

Up till now we can provide the following systems:

Confocal LIFA system, by spinning disk and modulated diode laser:
 
At the cameraport, the CSU10 or CSU22 spinning disk (e.g. Yokogawa distributor Perkin Elmer) is mounted, with the modulated intensified camera of the LIFA -Lambert Instruments Fluorescence Attachment - to the cameraport of the spinning disk. As light source the modulated diode laser is used, instead of the typical modulated LED light source. The modulation of the modulated diode laser is directed by the LIFA -Lambert Instruments Fluorescence Attachment, so still the LI FLIM software program is used to obtain confocal lifetime images.
Read more about the technology of confocal FLIM and the applications of confocal FLIM, and view the confocal fluorescence lifetime images.
Tested modulated diode lasers:
* 473 nm modulated diode laser of PicoQuant (this image),
* 445nm/473nm dual wavelength modulated diode laser of Omicron (integrated in the LIFA software package LI FLIM).

TIRF LIFA system, by white-TIRF (LED illumination) or by laser-TIRF (diode laser illumination):

In this image, the LIFA -Lambert Instruments Fluorescence Attachment is shown attached to the Nikon TE2000-U widefield microscope with white-TIRF illuminator (-Total Internal Reflection Fluorescence). At the cameraport the modulated intensified camera of the LIFA -Lambert Instruments Fluorescence Attachment is mounted and as light source the modulated LED is used, which is typical for the LIFA -Lambert Instruments Fluorescence Attachment. In the light path a conical diaphragm is placed to only illuminate the side part of the objective. On the TIRF-FLIM Technology page more information is given on the illumination during TIRF -total internal reflection fluorescence.

More accurate measurements are obtained by laser-TIRF (-Total Internal Reflection Fluorescence), because of the more powerful illumination. The only difference with the white-TIRF (-Total Internal Reflection Fluorescence) system is the illumination. The modulated LED is replaced by a modulated diode laser that is aligned such that it is suitable for TIRF -total internal reflection fluorescence. The laser-TIRF illumination systems can be provided from different brands and can be used with diode lasers of various wavelenghts like 445nm and 473nm. Read more about the TIRF-FLIM Technology  and the TIRF-FLIM applications, and view the TIRF fluorescence lifetime images.
Tested TIRF systems:
* Nikon white-TIRF
* Nikon laser-TIRF
* Leica laser-TIRF
* Olympus laser-TIRF