Cell Solutions on the 2100 Bioanalyzer
Cell Solutions on the 2100 Bioanalyzer
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Note: This is a review of the published article listed below. All information, quotes, figures, methods, and findings mentioned in this review are from that article, and are the property of its authors and/or the publication in which the article originally appeared.
Research requiring cell enumeration demands rapid, simple, and consistent methods for accurate characterization of cell populations. In the past, researchers have turned to techniques such as fluorescent staining and microscopic observation for cell enumeration, however, theses techniques are labor- and time-intensive and frequently result in inconsistent data. Flow cytometry offered an alternative to traditional techniques, but expense and maintenance requirements have limited the appeal of the associated instrumentation for many labs. The Agilent 2100 Bioanalyzer offers a cost-effective alternative to expensive instrumentation for simple flow cytometry applications, as well as all related reagents and solutions for sensitive and reproducible results. Chieko Sakamoto and researchers at Osaka University in Japan (2005) used the 2100 Bioanalyzer to perform rapid and simple quantification of bacterial cells. The group found that the 2100 Bioanalyzer-based analysis resulted in linear distribution of control beads and similar standard curves for both bacteria and large cells. Further, there was a strong correlation between the on-chip flow cytometric count and the epifluorescence microscopic count, yet on-chip technique offered the advantage of simultaneous enumeration of different cell types using fluorescent stains. Based on their results, the group suggests that the speed and ease of sample preparation enable the 2100 Bioanalyzer to excel for flow cytometry applications.
Figure 1. Standard curves with fluorescent beads analyzed by on-chip flow cytometry.
Shown are analyses of beads with a diameter of 1.7 µm (
), 1.0 µm (×), 0.7 µm (+), and 0.5 µm (—) (A), as well as beads with a diameter of 11 µm (
), 6.0 µm (□), 5 µm (
), and 2.8 µm (
) (B). The line indicates the linear regression.
Figure 2. Relationship between on-chip flow cytometric count (on-chip FCM) and epifluorescence microscopic count (EFM) of E. coli O157:H7.
Bacterial cells were stained with SYBR Green II (A) or double stained with SYBR Green II and TO-PRO 3 (B). On-chip flow cytometric counts were determined by the standard curve (Fig. 1A). The line represents the linear regression.
Figure 3. On-chip flow cytometric analysis.
On-chip flow cytometric analysis of E. coli O157:H7 (A), P. putida (B), a mixture of E. coli O157:H7 and P. putida (C), and sterilized deionized water (D) double stained by Alexa Fluor 647-labeled anti-E. coli direct antibody and SYBR Green II. The box labeled E. coli shows the area of E. coli O157:H7, and the P. putida box shows the area of P. putida.
Original Research Paper:
Title: Rapid and simple quantification of bacterial cells by using a microfluidic device.
Authors: Sakamoto C, Yamaguchi N, Nasu M.
Journal: Appl Environ Microbiol. 2005 Feb;71(2):1117-21.
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