Technical Reference #1818

1818.

Evolution of Recognition of Ligands from Gram-Positive Bacteria: Similarities and Differences in the TLR2-Mediated Response between Mammalian Vertebrates and Teleost Fish Carla M. S. Ribeiro; Trudi Hermsen; Anja J. Taverne-Thiele; Huub F. J. Savelkoul; and Geert F. Wiegertjes, Wageningen University, Journal of Immunology, 184(1818), (2010)
Link To Paper

Materials & Methods:
Confocal laser scanning microscopy Cytospin microscope slides of carp macrophages were made by fixing cells in 100%alcohol and37%formaldehyde (10:1).HEK293 cells were grownin 6- well glass bottom culture plate (MatTek AshlandMA)fixed in100%alcohol and 37% formaldehyde (10:1). Sections were embedded in Vectashield containing propidium iodide (PI) (Vector Laboratories BurlingameCA) and examined with a Zeiss LSM-510 laser scanning microscope. Green fluorescent signal (rhodamine or green-fluorescent protein) was excited with a 488-nm argon laser and detected using a band-pass filter (505–530 nm) and red-fluorescent signal (PI) was excited with a 543-nm helium-neon laser and detected using a long-pass filter (560 nm).

Microscopic Technique
Confocal laser scanning microscopy

Cell Type(s)
HECK293 cells