title: [Optimization of labeling and localizing bacterial membrane and nucleus with FM4-64 and Hoechst dyes].
abstract: OBJECTIVE
  To observe cell membrane and nucleus in bacteria for subcellular localization.
  
  
  METHODS
  FM4-64 and Hoechst were dyed that can label cell membrane and nucleus, respectively. Both dyes were used to co-stain the membranes and nucleus of eight bacterial strains ( Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Yersinia pestis, Legionella pneumonia, Vibrio cholerae and Bacillus anthracis). E. coli was dyed with different dye concentrations and times and then observed by confocal fluorescence microscopic imaging.
  
  
  RESULTS
  Fluorescence intensity of cell membrane and nucleus is affected by dye concentrations and times. The optimal conditions were determined as follows: staining cell membrane with 20 μg/mL FM4-64 for 1 min and cell nucleus with 20 μg/mL Hoechst for 20 min. Gram-negative bacteria were dyed better than gram-positive bacteria with FM4-64dye.
  
  
  CONCLUSIONS
  FM4-64 and Hoechst can be used to stain membrane and nucleus in different types of bacteria. Co-staining bacterial membrane and nucleus provides the reference to observe cell structure in prokaryotes for studying subcellular localization.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Yanping
authors.initials: Y
authors.lastname: Han
authors.forename: Ruifu
authors.initials: R
authors.lastname: Yang
authors.forename: Xingxu
authors.initials: X
authors.lastname: Zhao
mesh_terms: D001419:Bacteria / Q000737:chemistry*; D002462:Cell Membrane / Q000737:chemistry*; D002467:Cell Nucleus / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry; D011726:Pyridinium Compounds / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D013194:Staining and Labeling / Q000295:instrumentation / Q000379:methods*
chemical_list: C092350:FM 4-64; D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds