PiRamid: A compact Raspberry Pi imaging box to automate small-scale time-lapse digital analysis, suitable for laboratory and field use

Long, Matthew Michael; Diep, Tai The; Needs, Sarah; Ross, Marta J.; Edwards, Alexander D.

doi:10.1016/j.ohx.2022.e00377

Cited by 3 publications

(6 citation statements)

References 27 publications

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“…Some differences in calculated absorbances within individual capillaries were detected ( Figure 2 a,b). Surprisingly, although visually different, with the transillumination imaging setup used here [ 33 ], clear changes in red channel absorbance still indicated that metabolic conversion of resazurin is easy to quantify colourimetrically in the presence of milk. Only the highest milk concentrations (50% final concentration) gave a large enough increase in baseline absorbance to reduce the growth detection measurement range.…”

Section: Resultsmentioning

confidence: 97%

“…Microcapillary test strips were dipped into the wells and the strips sealed with silicone grease to avoid evaporation. Samples were incubated at 37 °C overnight, colour change was time-lapse imaged using Raspberry Pi-operated automated imaging devices PiRamid [ 33 ]. The MIC was determined based on the lowest concentration that inhibited the reduction in resazurin from blue to pink after overnight incubation.…”

Section: Methodsmentioning

confidence: 99%

“…All time-lapse imaging was carried out on a low-cost open-source imaging system designed for time-lapse imaging of colourimetric assays [ 33 ]. Image J was used to analyse the time-lapse image series of bacterial growth in the microcapillary test strips.…”

Section: Methodsmentioning

confidence: 99%

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Dilution Reduces Sample Matrix Effects for Rapid, Direct, and Miniaturised Phenotypic Antibiotic Susceptibility Tests for Bovine Mastitis

Long,

Needs,

Edwards

2023

Antibiotics

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The time-consuming nature of current methods for detecting antimicrobial resistance (AMR) to guide mastitis treatment and for surveillance, drives innovation towards faster, easier, and more portable technology. Rapid on-farm testing could guide antibiotic selection, reducing misuse that contributes to resistance. We identify challenges that arise when developing miniaturized antibiotic susceptibility tests (AST) for rapid on-farm use directly in milk. We experimentally studied three factors: sample matrix (specifically milk or spoiled milk); the commensal bacteria found in fresh bovine milk; and result time on the performance of miniaturised AST. Microfluidic “dip-and-test” devices made from microcapillary film (MCF) were able to monitor Gram-negative bacterial growth colourimetrically even in the presence of milk and yoghurt (used to simulate spoiled milk samples), as long as this sample matrix was diluted 1:5 or more in growth medium. Growth detection kinetics using resazurin was not changed by milk at final concentrations of 20% or lower, but a significant delay was seen with yoghurt above 10%. The minimum inhibitory concentration (MIC) for ciprofloxacin and gentamicin was increased in the presence of higher concentrations of milk and yoghurt. When diluted to 1% all observed MIC were within range, indicating dilution may be sufficient to avoid milk matrix interfering with microfluidic AST. We found a median commensal cell count of 6 × 105 CFU/mL across 40 healthy milk samples and tested if these bacteria could alter microfluidic AST. We found that false susceptibility may be observed at early endpoint times if testing some pathogen and commensal mixtures. However, such errors are only expected to occur when a susceptible commensal organism is present at higher cell density relative to the resistant pathogen, and this can be avoided by reading at later endpoints, leading to a trade-off between accuracy and time-to-result. We conclude that with further optimisation, and additional studies of Gram-positive organisms, it should be possible to obtain rapid results for microfluidic AST, but a trade-off is needed between time-to-result, sample dilution, and accuracy.

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Section: Resultsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

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Dilution Reduces Sample Matrix Effects for Rapid, Direct, and Miniaturised Phenotypic Antibiotic Susceptibility Tests for Bovine Mastitis

Long,

Needs,

Edwards

2023

Antibiotics

Self Cite

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“…Various camera modules are available from Raspberry Pi (Table 2, Figure 2). The Raspberry Pi Camera Module 2 (GBP 28.50) The Raspberry Pi computer/camera system has been used in conjunction with microfluidics for different biological studies, ranging from colour and fluorescence detection [37][38][39][40]; for direct cellular image detection and microscopy [39,[41][42][43][44][45][46]; for time-lapse imaging [47]; and for droplet control microfluidics [1,[48][49][50][51] (Table 3).…”

Section: Raspberry Pi/pi Camera Systemmentioning

confidence: 99%

“…The Raspberry Pi V2 camera module has been used in many systems [38,47,55,56]. A channel-based system used the Raspberry Pi V2 module to image colourimetric changes in 0.3 mm wide capillaries [7,55].…”

Section: Colour and Fluorescence Detection In Microfluidicsmentioning

confidence: 99%

Open Hardware for Microfluidics: Exploiting Raspberry Pi Singleboard Computer and Camera Systems for Customisable Laboratory Instrumentation

Sarıyer,

Edwards,

Needs

2023

Biosensors

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The integration of Raspberry Pi miniature computer systems with microfluidics has revolutionised the development of low-cost and customizable analytical systems in life science laboratories. This review explores the applications of Raspberry Pi in microfluidics, with a focus on imaging, including microscopy and automated image capture. By leveraging the low cost, flexibility and accessibility of Raspberry Pi components, high-resolution imaging and analysis have been achieved in direct mammalian and bacterial cellular imaging and a plethora of image-based biochemical and molecular assays, from immunoassays, through microbial growth, to nucleic acid methods such as real-time-qPCR. The control of image capture permitted by Raspberry Pi hardware can also be combined with onboard image analysis. Open-source hardware offers an opportunity to develop complex laboratory instrumentation systems at a fraction of the cost of commercial equipment and, importantly, offers an opportunity for complete customisation to meet the users’ needs. However, these benefits come with a trade-off: challenges remain for those wishing to incorporate open-source hardware equipment in their own work, including requirements for construction and operator skill, the need for good documentation and the availability of rapid prototyping such as 3D printing plus other components. These advances in open-source hardware have the potential to improve the efficiency, accessibility, and cost-effectiveness of microfluidic-based experiments and applications.

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Evaluating Factors Shaping Real-Time Internet-of-Things-Based License Plate Recognition Using Single-Board Computer Technology

Netinant,

Phonsawang,

Rukhiran

2024

Technologies

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Reliable and cost-efficient license plate recognition (LPR) systems enhance security, traffic management, and automated toll collection in real-world applications. This study addresses optimal unique configurations for enhancing LPR system accuracy and reliability by evaluating the impact of camera angle, object velocity, and distance on the efficacy of real-time LPR systems. The Internet of Things (IoT) LPR framework is proposed and utilized on single-board computer (SBC) technology, such as the Raspberry Pi 4 platform, with a high-resolution webcam using advanced OpenCV and OCR–Tesseract algorithms applied. The research endeavors to simulate common deployment scenarios of the real-time LPR system and perform thorough testing by leveraging SBC computational capabilities and the webcam’s imaging capabilities. The testing process is not just comprehensive, but also meticulous, ensuring the system’s reliability in various operational settings. We performed extensive experiments with a hundred repetitions at diverse angles, velocities, and distances. An assessment of the data’s precision, recall, and F1 score indicates the accuracy with which Thai license plates are identified. The results show that camera angles close to 180° significantly reduce perspective distortion, thus enhancing precision. Lower vehicle speeds (<10 km/h) and shorter distances (<10 m) also improve recognition accuracy by reducing motion blur and improving image clarity. Images captured from shorter distances (approximately less than 10 m) are more accurate for high-resolution character recognition. This study substantially contributes to SBC technology utilizing IoT-based real-time LPR systems for practical, accurate, and cost-effective implementations.

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PiRamid: A compact Raspberry Pi imaging box to automate small-scale time-lapse digital analysis, suitable for laboratory and field use

Cited by 3 publications

References 27 publications

Dilution Reduces Sample Matrix Effects for Rapid, Direct, and Miniaturised Phenotypic Antibiotic Susceptibility Tests for Bovine Mastitis

Dilution Reduces Sample Matrix Effects for Rapid, Direct, and Miniaturised Phenotypic Antibiotic Susceptibility Tests for Bovine Mastitis

Open Hardware for Microfluidics: Exploiting Raspberry Pi Singleboard Computer and Camera Systems for Customisable Laboratory Instrumentation

Evaluating Factors Shaping Real-Time Internet-of-Things-Based License Plate Recognition Using Single-Board Computer Technology

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