Digital postprocessing and image segmentation for objective analysis of colorimetric reactions

Woolf, M. Shane; Dignan, Leah M.; Scott, Anchi; Landers, James P.

doi:10.1038/s41596-020-00413-0

Cited by 72 publications

(80 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Quantification of matrix prevalence in the eluate was enabled by collecting empirical measurements of the hue, the circular variable that represents a color within the visible portion of the electromagnetic spectrum. 22 The hue of the yellow dye in the eluate chamber after the completion of the assay workflow (0.175 ± 0.003) was statistically significantly higher than that of pure yellow dye (0.147 ± 0.004), according to an unpaired t test for means (α = 0.05, two-tailed p -value <0.0001). This upward shift in hue was attributed to contamination with the blue dye (hue = 0.571 ± 0.002), which reflects the carryover of the matrix into the viral RNA eluate ( Figure 4 C).…”

Section: Resultsmentioning

confidence: 92%

“…Raw images were converted to HSB stacks using the Fiji ImageJ freeware and analyzed using the “crop-and-go” technique. 22 Briefly, the hue of a rectangular region of interest (ROI) within each chamber ( n = 6) and parallel measurements were compared to control dye solutions of known constituency on-disc. Control solutions were comprised of serially diluted blue dye in yellow dye from 10% to 0% (each n = 3).…”

Section: Materials and Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Centrifugal Microfluidic Method for Enrichment and Enzymatic Extraction of Severe Acute Respiratory Syndrome Coronavirus 2 RNA

et al. 2022

Self Cite

View full text Add to dashboard Cite

The diversification of analytical tools for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is imperative for effective virus surveillance and transmission control worldwide. Development of robust methods for rapid, simple isolation of viral RNA permits more expedient pathogen detection by downstream real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR) to minimize stalled containment and enhance treatment efforts. Here, we describe an automatable rotationally driven microfluidic platform for enrichment and enzymatic extraction of SARS-CoV-2 RNA from multiple sample types. The multiplexed, enclosed microfluidic centrifugal device (μCD) is capable of preparing amplification-ready RNA from up to six samples in under 15 min, minimizing user intervention and limiting analyst exposure to pathogens. Sample enrichment leverages Nanotrap Magnetic Virus Particles to isolate intact SARS-CoV-2 virions from nasopharyngeal and/or saliva samples, enabling the removal of complex matrices that inhibit downstream RNA amplification and detection. Subsequently, viral capsids are lysed using an enzymatic lysis cocktail for release of pathogenic nucleic acids into a PCR-compatible buffer, obviating the need for downstream purification. Early in-tube assay characterization demonstrated comparable performance between our technique and a “gold-standard” commercial RNA extraction and purification kit. RNA obtained using the fully integrated μCDs permitted reliable SARS-CoV-2 detection by real-time RT-PCR. Notably, we successfully analyzed full-process controls, positive clinical nasopharyngeal swabs suspended in viral transport media, and spiked saliva samples, showcasing the method’s broad applicability with multiple sample matrices commonly encountered in clinical diagnostics.

show abstract

Section: Resultsmentioning

confidence: 92%

Section: Materials and Methodsmentioning

confidence: 99%

Centrifugal Microfluidic Method for Enrichment and Enzymatic Extraction of Severe Acute Respiratory Syndrome Coronavirus 2 RNA

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Thus, employing analytical methods that identify products formation or the substrate consumption in a colored mixture could segregate the "EDA-hit" from colorimetric reactions (reactionbased screening). 27 In this study, we have explored EDA complex-mediated reactions, where the substrates (electron donor and acceptor) are mixed and submitted to visible-light irradiation to undergo chemical transformations. 5,6,8 Other approaches, which use sacri cial electron donor-or acceptor-species for EDA complex formation, and subsequent trapping of the target radical with a third substrate, 7,11 were not covered in this study.…”

Section: Resultsmentioning

confidence: 99%

Discovery of EDA-Complex Photocatalyzed Reactions Using Multidimensional Image Processing: Iminophosphorane Synthesis as a Case Study

Silva

Batista

Brocksom

et al. 2021

Preprint

View full text Add to dashboard Cite

Herein, we report a multidimensional screening strategy for the discovery of EDA-complex photocatalyzed reactions using only photographic devices (webcam, cellphone) and TLC analysis. An algorithm was designed to identify automatically EDA-complex reactive mixtures in solution from digital image processing in a 96-wells microplate and by TLC-analysis. The code highlights the region of absorption of the mixture in the visible spectrum, and the quantity of the color change through grayscale values. Furthermore, the code identifies automatically the blurs on the TLC plate and classifies the mixture as colorimetric reactions, non-reactive or potentially reactive EDA mixtures. This strategy allowed us to discover and then optimize a new EDA-mediated approach for obtaining iminophosphoranes in up to 90% yield.

show abstract

“…Visual detection with color-generating reactions may seem trivial, but depending on lighting, color blindness, age, and a number of other parameters, there is an inherent subjectivity, as it is well-established that no two individuals see color exactly the same way [17]. To alleviate subjectivity in the color analysis, we build a "lightbox" with static lighting along with a Huawei P9 smartphone (vendor) to be used for image acquisition [18] (Figure 2).…”

Section: Image Capture and Analysismentioning

confidence: 99%

Optimization of novel loop‐mediated isothermal amplification with colorimetric image analysis for forensic body fluid identification

Layne

Scott

Tanner

et al. 2021

Journal of Forensic Sciences

Self Cite

View full text Add to dashboard Cite

Body fluids are a category of biological evidence that provides critical contextual clues for forensic investigations; blood, saliva, and semen are three of the most prevalently tested fluids in a forensic laboratory. Some of the existing colorimetric presumptive forensic assays for these fluids include: the phenolphthalein test or luminol test for blood, the acid phosphatase (AP) test or prostate-specific antigen (PSA) lateral flow assay for seminal fluid, and the alpha-amylase test for saliva. Even though these enzymatic-and immunological-based tests are widely utilized in the forensic workflow, they often lack sensitivity and specificity required for a confirmatory assay [1,2]. For example, a study by

show abstract

Digital postprocessing and image segmentation for objective analysis of colorimetric reactions

Cited by 72 publications

References 29 publications

Centrifugal Microfluidic Method for Enrichment and Enzymatic Extraction of Severe Acute Respiratory Syndrome Coronavirus 2 RNA

Centrifugal Microfluidic Method for Enrichment and Enzymatic Extraction of Severe Acute Respiratory Syndrome Coronavirus 2 RNA

Discovery of EDA-Complex Photocatalyzed Reactions Using Multidimensional Image Processing: Iminophosphorane Synthesis as a Case Study

Optimization of novel loop‐mediated isothermal amplification with colorimetric image analysis for forensic body fluid identification

Contact Info

Product

Resources

About