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Rapid and effective methods for the detection of analytes such as water contaminants, food adulterants and biomolecules are essential for the protection of public health and environmental protection. Most of the currently established analytical techniques need sophisticated equipment, centralized testing facilities, costly operations, and trained personnel. Such limitations make them inaccessible to the general populace, particularly in regions with limited resources. The emergence of microfluidic devices offers a promising alternative to overcome several such constraints. This work describes a protocol for fabricating a low-cost, open-source paper-based microfluidic device using easily available tools and materials for colorimetric detection of analytes. The ease and simplicity of fabrication allow users to design customized devices. The device is coupled with an imaging box assembled from 3D printed parts to maintain uniform lighting conditions during analytical testing. The platform allows digital imaging using smartphones or cameras to instantaneously capture images of reaction zones on the device for quantitative analysis. The system is demonstrated for detecting hexavalent chromium, a toxic water contaminant. The image analysis is performed using open-source ImageJ for quantification of results. The approach demonstrated in this work can be readily adopted for a wide range of sensing applications., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)