研究目的
To develop and evaluate indicator-based polymeric color standards for digital color analysis of optical chemical sensors, aiming to improve accuracy and reliability compared to conventional gray standards.
研究成果
The developed indicator-based polymeric color standards provide stable and tunable colors independent of solution composition, offering improved accuracy, broader span, and higher sensitivity in digital color analysis for pH optodes compared to conventional gray standards. They are functional in complex media like serum and show promise for integration into high-throughput sensing platforms without additional fabrication methods.
研究不足
The study is limited to specific lipophilic indicators (ETH5350 and ETH2439) and electrolytes (TBATBB), and may not generalize to other materials. The optodes require controlled illumination and environmental conditions for accurate measurements. Long-term stability and photochemical degradation under UV light were not extensively tested, and applicability to other biological matrices beyond horse serum was not explored.
1：Experimental Design and Method Selection:
The study involved designing polymeric optode membranes containing lipophilic pH-indicators (ETH5350 and ETH2439), ionophores, ionic additives, and lipophilic electrolytes (TBATBB) to create color standards. The methodology included spectrophotometric analysis and digital color imaging to assess color stability and response.
2：Sample Selection and Data Sources:
Aqueous solutions with varying pH (4.6 to 12) and NaCl concentrations (0.01 M to 1 M), and diluted horse serum samples were used. Data were collected from replicate optode measurements.
3：6 to 12) and NaCl concentrations (01 M to 1 M), and diluted horse serum samples were used. Data were collected from replicate optode measurements.
List of Experimental Equipment and Materials:
3. List of Experimental Equipment and Materials: Digital camera MS-5 coupled with stereomicroscope MSP-2 (LOMO, Russia), micropipette (0.1–2.5 μL, Biohit, Finland), glass slides with microwells, incandescent lamps (Natural light, 60 W, Philips), ImageJ software, OriginPro 9.0 for data analysis. Chemicals included ETH5350, ETH2439, sodium ionophore VI, NaHFPB, TBATBB, PVC, DOS, THF, HEPES, inorganic salts, and horse serum.
4：1–5 μL, Biohit, Finland), glass slides with microwells, incandescent lamps (Natural light, 60 W, Philips), ImageJ software, OriginPro 0 for data analysis. Chemicals included ETH5350, ETH2439, sodium ionophore VI, NaHFPB, TBATBB, PVC, DOS, THF, HEPES, inorganic salts, and horse serum.
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Optode membranes were prepared by dissolving components in THF and casting into microwells. After solvent evaporation, membranes were photographed under controlled illumination after contact with samples. Images were processed in RGB color space, with signals referenced to standards. Calibration curves were generated for pH response.
5：Data Analysis Methods:
Optical signals (R, G, B components) were averaged using ImageJ. Referenced intensities (Zref = Zraw/Zst) and ratios (Rref/Bref) were calculated. Data were fitted with sigmoid or linear functions in OriginPro 9.0, with error analysis based on standard deviations.
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