An image cytometer based on angular spatial frequency processing and its validation for rapid detection and quantification of waterborne microorganisms

We introduce a new image cytometer design for the detection of very small particulates and demonstrate its capability in water analysis. The device is a compact microscope composed of off-the-shelf components, such as a light emitting diode (LED) source, a complementary metal-oxide-semiconductor (CM...

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Bibliographic Details
Published in:Analyst (London) 2015-11, Vol.14 (22), p.7734-7741
Main Authors: Pérez, J. M, Jofre, M, Martínez, P, Yáñez, M. A, Catalan, V, Pruneri, V
Format: Article
Language:English
Subjects:
Computer programs
Devices
Equipment Design
Escherichia coli
Escherichia coli - isolation & purification
Fluorescein-5-isothiocyanate - analysis
Fluorescent Dyes - analysis
Image Cytometry - economics
Image Cytometry - instrumentation
Image detection
Legionella pneumophila
Legionella pneumophila - isolation & purification
Light-emitting diodes
Microorganisms
Microscopy - instrumentation
Particulates
Semiconductors
Water analysis
Water Microbiology
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Summary:We introduce a new image cytometer design for the detection of very small particulates and demonstrate its capability in water analysis. The device is a compact microscope composed of off-the-shelf components, such as a light emitting diode (LED) source, a complementary metal-oxide-semiconductor (CMOS) image sensor, and a specific combination of optical lenses that allow, through an appropriate software, Fourier transform processing of the sample volume. Waterborne microorganisms, such as Escherichia coli ( E. coli ), Legionella pneumophila ( L. pneumophila ) and phytoplankton, are detected by interrogating the volume sample either in a fluorescent or label-free mode, i.e. with or without fluorescein isothiocyanate (FITC) molecules attached to the micro-organisms, respectively. We achieve a sensitivity of 50 cells per ml, which can be further increased to 0.2 cells per ml by pre-concentrating an initial sample volume of 500 ml with an ad hoc fluidic system. We also prove the capability of the proposed image cytometer of differentiating microbiological populations by size with a resolution of 3 μm and operating in real contaminated water. Image cytometer based on angular spatial frequency processing for the early detection of waterborne bacteria.
ISSN:0003-2654
1364-5528
DOI:10.1039/c5an01338k