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Simplified sample preparation using frame spotting method for direct counting of total bacteria by fluorescence
Fumito Maruyama1, Nobuyasu Yamaguchi, Takehiko Kenzaka
1Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6, Yamada-oka, Suita, Osaka 565-0871, Japan.
Journal of Microbiological Methods
|October 19, 2004
Summary
A novel method simplifies direct bacteria counting in liquid samples using a coated glass slide and polyester seal. This rapid, inexpensive technique offers reproducible results comparable to conventional fluorescence microscopy methods.
Area of Science:
- Microbiology
- Analytical Chemistry
- Microscopy
Background:
- Accurate enumeration of bacteria in liquid samples is crucial for various scientific and industrial applications.
- Conventional methods for bacterial counting, such as filtration coupled with fluorescence microscopy, can be time-consuming and resource-intensive.
Purpose of the Study:
- To develop and validate a simplified, rapid, and cost-effective method for the direct enumeration of bacteria in liquid samples using fluorescence microscopy.
Main Methods:
- A novel preparation technique involving a glass slide coated with 3-aminopropyltriethoxy silane and a ring-shaped polyester seal was employed.
- The method includes spotting samples, vacuum drying, staining with SYBR Green II, further drying, and mounting for direct microscopic counting.
Main Results:
- The developed method demonstrated consistent and reproducible bacterial counts when compared to the conventional filtration method.
- The simplified procedure significantly reduces the time and cost associated with bacterial enumeration.
Conclusions:
- This new preparation method offers a practical and efficient alternative for direct bacterial counting in liquid samples.
- The technique is suitable for routine laboratory use, providing reliable results for fluorescence microscopy applications.