Application of Photoshop and Scion Image analysis to quantification of signals in histochemistry, immunocytochemistry and hybridocytochemistry.
OBJECTIVE
To describe a simple method to achieve the differential selection and subsequent quantification of the strength signal using only one section.


METHODS
Several methods for performing quantitative histochemistry, immunocytochemistry or hybridocytochemistry, without use of specific commercial image analysis systems, rely on pixel-counting algorithms, which do not provide information on the amount of chromogen present in the section. Other techniques use complex algorithms to calculate the cumulative signal strength using two consecutive sections. To separate the chromogen signal we used the "Color range" option of the Adobe Photoshop program, which provides a specific file for a particular chromogen selection that could be applied on similar sections. The measurement of the chromogen signal strength of the specific staining is achieved with the Scion Image software program.


CONCLUSIONS
The method described in this paper can also be applied to simultaneous detection of different signals on the same section or different parameters (area of particles, number of particles, etc.) when the "Analyze particles" tool of the Scion program is used.
Tolivia
Jorge
J
Departamento de Morfología y Biología Celular, Facultad de Biología y Medicina, Universidad de Oviedo, Spain. jtolivia@uniovi.es
Navarro
Ana
A
del Valle
Eva
E
Perez
Cristina
C
Ordoñez
Cristina
C
Martínez
Eva
E
D000818:Animals; D001053:Apolipoproteins / Q000378:metabolism; D053399:Apolipoproteins D; D001943:Breast Neoplasms / Q000378:metabolism; D002863:Chromogenic Compounds / Q000032:analysis*; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012984:Software
D001053:Apolipoproteins; D053399:Apolipoproteins D; D002863:Chromogenic Compounds; D012333:RNA, Messenger