Quantitative analysis of turbellarian cell suspension by fluorescent staining with acridine orange, and video microscopy
Behensky, C.; Schürmann, W.; Peter, R. (2001). Quantitative analysis of turbellarian cell suspension by fluorescent staining with acridine orange, and video microscopy. Belg. J. Zool. 131(Suppl. 1): 131-136
In: Belgian Journal of Zoology. Koninklijke Belgische Vereniging voor Dierkunde = Société royale zoologique de Belgique: Gent. ISSN 0777-6276; e-ISSN 2295-0451, more
Also appears in:Saló, E.; Watson, N.; Schockaert, E. (Ed.) (2001). Proceedings of the 9 th International Symposium on the Biology of the Turbellaria, Barcelona, Spain, June 2000. Belgian Journal of Zoology, 131(Suppl. 1). Koninklijke Belgische Vereniging voor Dierkunde = Société royale zoologique de Belgique: Diepenbeek. 236 pp., more
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| Keywords |
Acids > Organic compounds > Organic acids > Nucleic acids > DNA
Acids > Organic compounds > Organic acids > Nucleic acids > RNA
Analysis > Image analysis
Analytical techniques > Microscopy > Fluorescence microscopy
Biological phenomena > Regeneration
Cell constituents > Chromosomes > Genes
Cultures > Cell suspensions
Image analysis
Image interpretation > Image analysis
Morphogenesis
Toxicology > Toxic substances > Mutagens > Acridines > Acridine orange
Macrostomum Schmidt, 1848 [WoRMS]; Platyhelminthes [WoRMS]
Marine/Coastal |
| Authors | | Top |
- Behensky, C.
- Schürmann, W.
- Peter, R.
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| Abstract |
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. |
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