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

TL;DR

This paper presents a compact, low-cost image cytometer utilizing angular spatial frequency processing for rapid detection and quantification of waterborne microorganisms, demonstrating high sensitivity and the ability to differentiate microbial populations.

Contribution

The study introduces a novel, portable image cytometer design that combines Fourier transform processing with off-the-shelf components for water analysis.

Findings

Sensitivity of 50 cells/ml, improved to 0.2 cells/ml with preconcentration

Differentiates microorganisms by size with 3 um resolution

Operates effectively in real contaminated water samples

Abstract

We introduce a new image cytometer design for detection of very small particulate 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/ml, which can be further increased to 0.2 cells/ml by preconcentrating an initial sample volume of 500 ml with an adhoc fluidic system. We also prove the capability of the proposed image cytometer of differentiating microbiological populations by size with a resolution of 3 um and of operating in real contaminated water.