Confocal Lifetime Images on a Widefield Fluorescence Microscope

Leutingewolde, The Netherlands - Nov. 21, 2006…Lambert Instruments announced today the capabiltiy to obtain confocal lifetime images on a widefield fluorescence microscope using the Lambert LIFA System.

The Lambert LIFA is used together with a fluorescence widefield microscope and a CSU10 spinning disk from Perkin Elmer/Yokogawa. The intensified CCD camera of the LIFA is used as detector by mounting it on the camera port of the CSU10 spinning disk. In this set-up, the light source is not the modulated LED typically used, but instead a modulated 470nm laser diode (PicoQuant). The rest of the set-up uses the standard LIFA components.

The confocal scanning method of the spinning disk is based on the Nipkow disk scanner - an optical scanner using rotation of a disk with pinholes to produce an image. By placing a microlens array in front of the Nipkow disk, the optical efficiency is improved about two orders of magnitude. More details about the technology can be found on the Yokogawa website.

Shown below are two z-positions FLIM maps taken for the same pollen grain sample. The acquisition was done with 12 phase steps of 50 msec exposure time at an MCP voltage of 690V. These two images are shown as a merge of the fluorescence lifetime in pseudo colours and the intensity in gray scale.

Please contact Advanced Labs for more information.

Tags: fluorescence, widefield microscope, FLIM