Title:Confocal Laser Scanning Microscope, Raman Microscopy and Western Blotting to Evaluate Inflammatory Response after Myocardial Infarction
VOLUME: 13 ISSUE: 1
Author(s):Irene Riezzo, Santina Cantatore, Dania De Carlo, Carmela Fiore, Margherita Neri, Emanuela Turillazzi and Vittorio Fineschi
Affiliation:Department of Forensic Pathology, Ospedale Colonnello D'Avanzo, Viale degli Aviatori 1, 71100 Foggia, Italy.
Keywords:Confocal laser microscope, cytokines, myocardial infarction, Raman microscope, western blotting.
Abstract:Cardiac muscle necrosis is associated with inflammatory cascade that clears the infarct from dead
cells and matrix debris, and then replaces the damaged tissue with scar, through three overlapping phases: the
inflammatory phase, the proliferative phase and the maturation phase.
Western blotting, laser confocal microscopy, Raman microscopy are valuable tools for studying the inflammatory
response following myocardial infarction both humoral and cellular phase, allowing the identification and
semiquantitative analysis of proteins produced during the inflammatory cascade activation and the topographical distribution
and expression of proteins and cells involved in myocardial inflammation. Confocal laser scanning microscopy
(CLSM) is a relatively new technique for microscopic imaging, that allows greater resolution, optical sectioning of the
sample and three-dimensional reconstruction of the same sample. Western blotting used to detect the presence of a specific
protein with antibody-antigen interaction in the midst of a complex protein mixture extracted from cells, produced
semi-quantitative data quite easy to interpret. Confocal Raman microscopy combines the three-dimensional optical resolution
of confocal microscopy and the sensitivity to molecular vibrations, which characterizes Raman spectroscopy.
The combined use of western blotting and confocal microscope allows detecting the presence of proteins in the sample
and trying to observe the exact location within the tissue, or the topographical distribution of the same. Once demonstrated
the presence of proteins (cytokines, chemokines, etc.) is important to know the topographical distribution, obtaining in this
way additional information regarding the extension of the inflammatory process in function of the time stayed from the
time of myocardial infarction. These methods may be useful to study and define the expression of a wide range of inflammatory
mediators at several different timepoints providing a more detailed analysis of the time course of the infarct.
