User:Pakpoom Subsoontorn/Notebook/Shape Determination in Bacteria/2008/10/13

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Cell and membrane stainings

  • Here are the list of staining dyes we used with some detail

FM4-64

  • target:membrane staining dye
  • Filter
    • absorption range:
    • emission range:

NAO (10-N-Nonyl Acridine Orange)

  • target: cardiolipin (CL) staining dye
  • Filter
    • absorption range:
    • emission range:
  • Chemistry:
    • NH group in position 10 of acridine orange is replaced by N-nonyl group, increasing hydrophobicity and preventing h-bond formation with DNA or RNA. Binding to anionic phospholipid on membrane due to interaction between dye's quaternary amine and membrane phosphate residue. As CL has two phosphate group, the dye molecules are dimerized on the CL.
    • CL affinity to NAO, Ka ~ 2x10^6 /M...much higher than that of monoacidic acide, Ka~7x10^4/M

Reference

  • Mileykovskaya, E and Dowhan, W. Visaulization of Phospholipid domain in Escherichia coli by using the Caridolipin specific Fluorescence Dye 10-N-Nonyl Acridine Orange, Journal of Bacteriology, Feb 2000, p.1172-1175