研究目的
To develop miniaturized pH holographic sensors for non-invasive, real-time monitoring of Lactobacillus casei Shirota growth in a microfluidic chip, addressing the limitations of existing bioreactor monitoring techniques at small scales.
研究成果
The miniaturized pH holographic sensors successfully monitored L. casei growth in a microfluidic chip with accuracy comparable to conventional pH meters. This non-consumptive, optical approach enables real-time, non-invasive monitoring, potentially extending to arrays for multiple analytes and applications in diagnostics and wearables.
研究不足
The miniaturization of sensors requires high signal-to-noise ratio and uniform holographic signals. The fabrication process is sensitive to UV intensity and collimation, and non-specific polymerization can occur. The sensors have a narrow angular window for signal detection, necessitating precise alignment. Ionic strength variations in the medium could affect pH measurements, though efforts were made to normalize this.
1:Experimental Design and Method Selection:
The study involved fabricating miniaturized pH holographic sensors using photolithography and holographic techniques. The sensors were based on a pH-sensitive hydrogel with embedded silver nanoparticles, and their response was monitored through changes in diffracted wavelength due to gel volume changes.
2:Sample Selection and Data Sources:
Lactobacillus casei Shirota was used as the model organism, cultured in MRS broth. Samples were taken at various time points for pH, optical density, and conductivity measurements.
3:List of Experimental Equipment and Materials:
Materials included HEMA, EDMA, TFMPA, HQ, AgClO4, DMPA, Metol, L-ascorbic acid, lithium bromide, photomasks, spacers, COC covers, optomechanical apparatus, pH and conductivity meters, monochromator, spectrophotometer, high-resolution camera, and white light interferometric profiler. Equipment was sourced from various suppliers such as Merck, Acros Organics, Mettler Toledo, PhotoniTech, Knight Photonics, iSolutions Technology, and Bruker.
4:Experimental Procedures and Operational Workflow:
The hydrogel was polymerized on a silanized glass slide using UV exposure through photomasks, followed by hologram fabrication with a 532 nm laser. The sensors were integrated into a PDMS-glass microfluidic chip. Sensor interrogation involved a customized setup with a Xenon light source, fiber optics, CMOS camera, and spectrometer to measure the holographic signal. Calibration was done using McIlvaine buffers, and L. casei growth was monitored by introducing aliquots into the chip.
5:Data Analysis Methods:
Data analysis included normalization of wavelength spectra, application of the Henderson-Hasselbalch equation for pH prediction, and statistical analysis of sensor responses. Color changes were analyzed using RGB-to-xyY transformations and plotted on CIE chromaticity diagrams.
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pH meter
Mettler Toledo
Measuring pH of samples for comparison with sensor readings
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conductivity meter
Mettler Toledo
Measuring ionic strength of buffers and samples
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monochromator
PhotoniTech
Part of spectrophotometer setup for optical density measurements
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spectrophotometer
Avaspec
Knight Photonics
Measuring optical density and holographic signals
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high-resolution camera
iSolutions Technology
Capturing images of holographic sensors for color analysis
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white light interferometric 3D surface profiler
Bruker
Obtaining 3D digital elevation models of sensors
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Xenon light source
Providing white light for sensor interrogation
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fiber optic cable
Transmitting light to and from the sensor
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CMOS camera
Capturing holographic signals and images
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spectrometer
Measuring wavelength spectra of holographic signals
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532 nm laser
Used in hologram fabrication process
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UV source
For photo-polymerization of hydrogels
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