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Low-cost, open-access quantitative phase imaging of algal cells using the transport of intensity equation.

ABSTRACT: Phase microscopy allows stain-free imaging of transparent biological samples. One technique, using the transport of intensity equation (TIE), can be performed without dedicated hardware by simply processing pairs of images taken at known spacings within the sample. The resulting TIE images are quantitative phase maps of unstained biological samples. Therefore, spatially resolved optical path length (OPL) information can also be determined. Using low-cost, open-source hardware, we applied the TIE to living algal cells to measure their effect on OPL. We obtained OPL values that were repeatable within species and differed by distinct amounts depending on the species being measured. We suggest TIE imaging as a method of discrimination between different algal species and, potentially, non-biological materials, based on refractive index/OPL. Potential applications in biogeochemical modelling and climate sciences are suggested.

SUBMITTER: Grant SD

PROVIDER: S-EPMC7029887 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Low-cost, open-access quantitative phase imaging of algal cells using the transport of intensity equation.

Grant Stephen D SD Richford Kyle K Burdett Heidi L HL McKee David D Patton Brian R BR

Royal Society open science 20200129 1

Phase microscopy allows stain-free imaging of transparent biological samples. One technique, using the transport of intensity equation (TIE), can be performed without dedicated hardware by simply processing pairs of images taken at known spacings within the sample. The resulting TIE images are quantitative phase maps of unstained biological samples. Therefore, spatially resolved optical path length (OPL) information can also be determined. Using low-cost, open-source hardware, we applied the TIE ...[more]

PMID: 32218984

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Project description:BackgroundRemote monitoring of plants using hyperspectral imaging has become an important tool for the study of plant growth, development, and physiology. Many applications are oriented towards use in field environments to enable non-destructive analysis of crop responses due to factors such as drought, nutrient deficiency, and disease, e.g., using tram, drone, or airplane mounted instruments. The field setting introduces a wide range of uncontrolled environmental variables that make validation and interpretation of spectral responses challenging, and as such lab- and greenhouse-deployed systems for plant studies and phenotyping are of increasing interest. In this study, we have designed and developed an open-source, hyperspectral reflectance-based imaging system for lab-based plant experiments: the HyperScanner. The reliability and accuracy of HyperScanner were validated using drought and salt stress experiments with Arabidopsis thaliana.ResultsA robust, scalable, and reliable system was created. The system was built using open-sourced parts, and all custom parts, operational methods, and data have been made publicly available in order to maintain the open-source aim of HyperScanner. The gathered reflectance images showed changes in narrowband red and infrared reflectance spectra for each of the stress tests that was evident prior to other visual physiological responses and exhibited congruence with measurements using full-range contact spectrometers.ConclusionsHyperScanner offers the potential for reliable and inexpensive laboratory hyperspectral imaging systems. HyperScanner was able to quickly collect accurate reflectance curves on a variety of plant stress experiments. The resulting images showed spectral differences in plants shortly after application of a treatment but before visual manifestation. HyperScanner increases the capacity for spectroscopic and imaging-based analytical tools by providing more access to hyperspectral analyses in the laboratory setting.

Dataset Information

Low-cost, open-access quantitative phase imaging of algal cells using the transport of intensity equation.

Publications

Low-cost, open-access quantitative phase imaging of algal cells using the transport of intensity equation.

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