The Leica TCS SP2 system was designed for high resolution confocal image acquisition using laser point scanning of living and fixed specimens. This system is ideal for fixed specimens that require severe background suppression and cross-talk correction as well as detecting far red fluorescence not detectable by eye. The system also has 3D reconstruction capabilities as well as simultaneous multi-spectral scanning and image acquisition.

Technical Specifications

Applications

Some useful applications of a confocal system include but not limited to: acquisition of high quality fluorescence images of a single optical slice, acquisition of images from thick section s requiring severe background suppression, reconstruction of 3-D images, co-localization and expression studies, kinetic studies (FRAP).

Examples

Mouse Kidney: single optical slice Alexa Fluor-488 wheat germ agglutinin, a green-fluorescent lectin, was used to label glomeruli and convoluted tubules. The filamentous actin prevalent in glomeruli and the brush border were stained with red-fluorescent Alexa Fluor-568 phalloidin. Molecular Probes: FluoCells slide #3

Mouse Kidney Vasculature: compressed 3-D image RenCre transgenic mice were crossed with mice heterozygous for the R26R-Confetti conditional allele, with a CAG promoter, loxP site, and STOP cassette preventing transcription of the downstream Brainbow 2.1 sequences. Introduction of the renin-driven Cre recombinase results in recombination-dependent expression of GFP, YFP, CFP or RFP.
Courtesy of Sean Glenn (Dr. K. Gross Lab)

Simultaneous Scan of 3 fluorochromes: Co-localization (Molecular Probes FocalCheck Slide #1)

488nm 543nm 633nm overlay
488nm 543nm 633nm Overlay of 488, 543, 633nm
fluorchromes

(co-localization of 3 fluorchromes)

 

Confocal System