Author/Editor		Llopiz, Diana; Santamaria, Luis; Roman, Jose; Villaro, Ana-Cristina
Title		Measuring volume fractions of immunostaining: a comparison between two quantitative methods
Type		članek
Source		Image Anal Stereol
Vol. and No.		Letnik 20, št. Suppl 1
Publication year		2001
Volume		str. 175-80
Language		eng
Abstract		In the present study, our efforts were concentrated on the quantification of volume fraction of immunostaining (Vv) by the application of two quantitative stereological methods: area measurements with image analysis (IA) and point counting method (PCM). For this purpose, proadrenomedullin N-20 terminal peptide (PAMP) was chosen, which is a hypotensive molecule present in various tissues, including the pituitary gland. As it is well known, IA and PCM consist of different procedures to obtain the correspondent measurements. The measurements of Vv of PAMP immunostaining obtained by both methods were compared and the statistical analysis was made. These measurements were estimated on two different groups of paraffin sections: 20 and 3 microm sections. When 20 microm sections were measured, the IA technique gave overestimated values of Vv with respect to those obtained by PCM. The same study was carried out in sections with 3 microm in thickness. In this case the Vv values obtained for PAMP immunostaining were very similar. In summary, both PCM and IA can render similar results if 3 microm sections are used and the object analyzed has an uncomplicated morphology. In thick sections, however, IA can result in an overestimation of Vv; in this case PCM, although a more complex method, should always be used.
Descriptors		PITUITARY GLAND IMAGE PROCESSING, COMPUTER-ASSISTED RATS IMMUNOHISTOCHEMISTRY GONADOTROPINS, PITUITARY