研究目的
Investigating the use of cellulose derivative films as insulating materials or in composite electrodes for hot electron-induced electrochemiluminescence (HECL) and their application in bioaffinity assays.
研究成果
Cellulose derivative-based carbon black composite electrodes are effective for HECL excitation of various labels and can be used in bioaffinity assays like immunoassays, offering environmental benefits and good analytical performance with low detection limits and wide linear ranges.
研究不足
The adhesion of some cellulose derivatives to metal substrates was insufficient, leading to peeling or cracking; optimization of film thickness and manufacturing reproducibility was challenging; some electrodes showed variability in HECL performance.
1:Experimental Design and Method Selection:
The study involved preparing cellulose derivative films and composite electrodes with carbon black, using spin coating for electrode fabrication, and employing HECL measurements with a coulostatic pulse generator and photon counting for detection.
2:Sample Selection and Data Sources:
Cellulose derivatives (CA, CAP, CAB, EC) and carbon black were purchased from Sigma-Aldrich and Cabot; luminophores like Tb(III) chelate, Ru(bpy)3 2+, and fluorescein were used.
3:List of Experimental Equipment and Materials:
Spin coater (WS-650MZ-23NPPB, Laurell Technologies Corporation), SEM (Zeiss Sigma VP), AFM (Veeco Dimension 3100), coulostatic pulse generator (in-laboratory-built), photomultiplier tube (PerkinElmer MH1993), photon counter (SR400, Stanford Research System), spectrograph (Oriel Instruments MS125), EMCCD (Andor Technology DV465C-FI).
4:Experimental Procedures and Operational Workflow:
Electrodes were spin-coated onto brass or steel substrates, HECL measurements were conducted in a Teflon cell with specific pulse parameters, and immunoassays involved antibody coating and detection steps.
5:Data Analysis Methods:
Time-resolved HECL measurements, calibration curves, and statistical analysis of emission intensities and standard deviations.
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Scanning Electron Microscope
Zeiss Sigma VP
Zeiss
Used for characterizing the surface morphology of EC-CB and CAP-CB composite films.
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Atomic Force Microscope
Veeco Dimension 3100
Veeco
Used for characterizing the surface roughness of composite films in tapping mode.
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Photomultiplier Tube
MH1993
PerkinElmer
Used for optical detection of HECL emissions with interference filters.
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Photon Counter
SR400
Stanford Research System
Used for counting photons in HECL measurements, connected to a DC-300 MHz amplifier.
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EMCCD
DV465C-FI
Andor Technology
Used for measuring HECL emission spectra, attached to a spectrograph.
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Cellulose Acetate
SKU 419028
Sigma-Aldrich
Used as an insulating material in electrode films.
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Cellulose Acetate Propionate
SKU 340642
Sigma-Aldrich
Used as an insulating material in electrode films.
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Cellulose Acetate Butyrate
SKU 419052
Sigma-Aldrich
Used as an insulating material in electrode films.
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Ethyl Cellulose
SKU 247499
Sigma-Aldrich
Used as an insulating material in electrode films.
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Spin Coater
WS-650MZ-23NPPB
Laurell Technologies Corporation
Used for spin coating cellulose derivative films onto metal substrates to create electrodes.
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Multichannel Scaler
MCS-II
Nucleus
Used for multiscale data acquisition in HECL measurements.
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Spectrograph
MS125
Oriel Instruments
Used with EMCCD for spectral analysis of HECL emissions.
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Ultrasonic Homogenizer
Cole-Parmer
Used for mixing cellulose derivative and carbon black solutions.
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Carbon Black
Vulcan XC72
Cabot
Used as a conducting particle in composite electrodes to facilitate electron transport.
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