A rapid cell counting method utilising acridine orange as a novel discriminating marker for both cultured astrocytes and microglia

Journal of Neuroscience Methods

Volumn 165, Issue 2, 2007, Pages 223-229

  Lovelace, Michael D ^a   Cahill, David M ^a  

^a DEAKIN UNIVERSITY   (Australia)

Author keywords

Acridine orange; Astrocyte; Cell counting; Cell culture; Fluorescence microscopy; Lysosome; Microglia

Indexed keywords

ACRIDINE ORANGE; CELL NUCLEUS DNA; LEUCINE METHYL ESTER; RNA;

ANIMAL CELL; ANIMAL TISSUE; APOPTOSIS; ARTICLE; ASTROCYTE; CELL COUNT; CELL GROWTH; CELL STRUCTURE; CELL TYPE; CONTROLLED STUDY; FEMALE; FLUORESCENCE MICROSCOPY; HOMEOSTASIS; IMMUNE RESPONSE; LYSOSOME; MICROGLIA; NEWBORN; NONHUMAN; PRIORITY JOURNAL; RAT;

ACRIDINE ORANGE; ANIMALS; ANIMALS, NEWBORN; ASTROCYTES; CELL COUNT; CELLS, CULTURED; CENTRAL NERVOUS SYSTEM; COLORING AGENTS; FEMALE; LYSOSOMES; MICROGLIA; MICROSCOPY, FLUORESCENCE; NUCLEIC ACIDS; RATS; RATS, SPRAGUE-DAWLEY; STAINING AND LABELING; TIME FACTORS;

EID: 34547670635     PISSN: 01650270     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.jneumeth.2007.06.009     Document Type: Article

References (18)

1. Aschner M. Astrocytes as mediators of immune and inflammatory responses in the CNS. Neurotoxicology 19 (1998) 269-282
2. Darzynkiewicz Z., and Juan G. Differential staining of DNA and RNA. In: Robinson J.P., Darzynkiewicz Z., Dobrucki J., Hyun W., Orfao A., and Rabinovitch P. (Eds). Current protocols in cytometry (2005), John Wiley and Sons, New Jersey, USA Unit 7.3
3. De Vellis J., and Morrison R.S. Growth of purified astrocytes in a chemically defined medium. Proc Natl Acad Sci USA 78 (1981) 7205-7209
4. DrBio Webb Research Group (Cornell University). Acridine orange. http://www.drbio.cornell.edu/Bio-Optical/exo_BMW/fl_labels/GranpH/AO.htm; 2006.
5. Giulian D., and Baker T.J. Characterisation of ameboid microglia isolated from developing mammalian brain. J Neurosci 6 (1986) 2163-2178
6. Kalmar B., Kittel A., Lemmens R., Kornyei Z., and Madarasz E. Cultured astrocytes react to LPS with increased cyclooxygenase activity and phagocytosis. Neurochem Int 38 (2001) 453-461
7. Kobayashi Y., Uchimoto T., Nohara H., Kamehara R., Iwamura M., and Watanabe N. Mechanism of apoptosis induced by a lysosomotropic agent, l-leucyl-l-leucine methyl ester. Apoptosis 4 (1999) 357-362
8. Lovelace M. Stephenson G and Watson TG. Steroid 21-hydroxylase expression in cultured rat astrocytes. Brain Res Bull 61 (2003) 609-615
9. Lovelace M. Steroid metabolism in mammalian cultured astrocytes (2006), School of Life and Environmental Sciences, Deakin University, Ph.D Thesis Geelong, Victoria, Australia p. 357
10. Naveilhan P., Sargent-Tanguy S., Chagneau C., and Neveu I. Fluorescent activated cell sorting (FACS): a rapid and reliable method to estimate the number of neurons in a mixed population. J Neurosci Meth 129 (2003) 73-79
11. Olsson G., Brunmark A., and Brunk U.T. Acridine orange-mediated photodamage of microsomal and lysosomal fractions. Virchows Arch B 56 (1989) 247-257
12. Olsson G., Roberg K., and Rundquist I. The use of acridine orange cytofluorometry in the study of macrophage lysosomal exocytosis. Anal Cell Pathol 2 (1990) 179-188
13. Olsson G., Zdolsek J.M., and Brunk U.T. Photooxidative damage to lysosomes of cultured macrophages by acridine orange. Photochem Photobiol 51 (1990) 67-76
14. Renau-Piqueras J., Lindo L., Iborra F.J., Azorin I., and Guerri C. Analysis of the endocytic-lysosomal system (vacuolar apparatus) in astrocytes during proliferation and differentiation in primary culture. Int J Dev Biol 37 (1993) 565-572
15. Thiele D., and Lipsky P.E. Apoptosis is induced in cells with cytolytic potential by l-leucyl-l-leucine methyl ester. J Immunol 148 (1992) 3950-3957
16. Zimmer J., Castellano B., Gonzalez B., Jensen M.B., Pedersen E.B., and Finsen B.R. A double staining technique for simultaneous demonstration of astrocytes and microglia in brain sections and astroglial cell cultures. J Histochem Cytochem 39 (1991) 561-568
17. Zimmer J., Dalmau I., Castellano B., Pedersen E.B., Finsen B., and Gonzalez B. Reduction of the microglial cell number in rat primary glial cell cultures by exogenous addition of dibutyryl cyclic adenosine monophosphate. J Neuroimmunol 70 (1996) 123-129
18. Zwain I., and Yen S.S.C. Dehydroepiandrosterone: biosynthesis and metabolism in the brain. Endrocrinology 140 (1999) 880-887