Simplified sample preparation using frame spotting method for direct counting of total bacteria by fluorescence microscopy.
A new preparation method for direct counting of bacteria in liquid samples with fluorescence microscope was developed using a glass slide coated with 3-aminopropyltriethoxy silane and ring-shaped polyester seal as a retainer. The experimental steps of this method were spotting samples onto the coated slides with the seal, drying under vacuum, staining with SYBR Green II, drying and covering with immersion oil and coverslip to allow counting. This simplified method provided consistent results when compared with the conventional filtration method for fluorescence microscopy, and is rapid, inexpensive and reproducible.
Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6, Yamada-oka, Suita, Osaka 565-0871, Japan.
Fumito
F
Maruyama
Nobuyasu
N
Yamaguchi
Takehiko
T
Kenzaka
Katsuji
K
Tani
Masao
M
Nasu
D001431:Bacteriological Techniques / Q000379:methods; D015169:Colony Count, Microbial; D048168:Escherichia coli K12 / Q000302:isolation & purification*; D008856:Microscopy, Fluorescence / Q000379:methods; D009930:Organic Chemicals / Q000737:chemistry; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000379:methods*; D013194:Staining and Labeling; D014871:Water Microbiology
D009930:Organic Chemicals; C098798:SYBR Green II