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You are here: Home / Associated publications / Belgian Journal of Zoology / Bibliographic References / Quantitative analysis of turbellarian cell suspensions by fluorescent staining with acridine orange, and video microscopy

C Behensky, W Schurmann and R Peter (2001)

Quantitative analysis of turbellarian cell suspensions by fluorescent staining with acridine orange, and video microscopy

BELGIAN JOURNAL OF ZOOLOGY, 131(S1):131-136.

A combination of methods has been developed for analyzing cell suspensions from turbellarians with respect to cytochemical and morphometric parameters and with special emphasis on the characterization of neoblasts. Tissues were disintegrated by mechanical and enzymatic means. Neoblasts were separated and/or fractionated by different centrifugation protocols in Percoll density-gradients. Staining with acridine orange under conditions denaturing RNA but leaving intact double stranded DNA yielded green fluorescence of DNA and red phosphorescence of RNA. Both light emissions were documented and quantitative image analyses were performed. By computing the integral intensities of green and red light, quantitative measures were gained for DNA and RNA contents. By correlating these light emissions to each other or to cell size, histograms characteristic for given neoblast pools were obtained. The protocol described should facilitate monitoring the heterogeneity of the neoblast compartment as well as studying cellular dynamics during growth, regeneration and other physiological processes.

acridine orange; cell suspensions; disintegration of tissues; DNA; Dugesia tahitiensis; fluorescence microscopy; image analysis; Macrostomum; neoblast fractions; planarians; RNA; video microscopy; Platyhelminthes
9th International Symposium on the Biology of the Turbellaria (ISBT), BARCELONA, SPAIN, JUN, 2000
BJZ

ISSN 2295-0451 (online version)
ISSN 0777-6279 (printed version)
impact factor 2015: 0,87.

Editor-in-Chief:
Prof. Dr. Isa Schön
Royal Belgian Institute of Natural Sciences
Vautierstraat 29
1000 Brussels, Belgium

 



1863-1903
Annales de la Société malacologique de Belgique
 
1903-1923
​Annales de la Société royale malacologique et zoologique de Belgique
 
1923-1989
Annales de la Société Royale Zoologique de Belgique
 
1989-
Belgian Journal of Zoology