Clinical Chemistry

4.

Analytical Techniques and Instrumentation

4.1.

Spectrophotometry

4.1.1.

Basic Principles

4.1.1.1.

Electromagnetic Radiation

4.1.1.2.

Absorbance and Transmittance

4.1.1.3.

Wavelength Selection

4.1.1.4.

Optical Density

4.1.2.

Beer-Lambert Law

4.1.2.1.

Mathematical Expression

4.1.2.2.

Limitations and Deviations

4.1.2.3.

Practical Applications

4.1.3.

Instrumentation

4.1.3.1.

4.1.3.1.1.

4.1.3.1.2.

Deuterium Lamps

4.1.3.1.3.

4.1.3.1.4.

Maintenance and Calibration

4.1.3.2.

4.1.3.2.1.

4.1.3.2.2.

Diffraction Gratings

4.1.3.2.3.

4.1.3.2.4.

Function and Adjustment

4.1.3.2.5.

4.1.3.3.

Sample Cell (Cuvette)

4.1.3.3.1.

Materials and Path Length

4.1.3.3.2.

Cleaning and Handling

4.1.3.3.3.

Temperature Control

4.1.3.4.

4.1.3.4.1.

4.1.3.4.2.

Photomultiplier Tubes

4.1.3.4.3.

Signal Processing

4.1.3.4.4.

Noise Reduction

4.1.4.

Types of Spectrophotometry

4.1.4.1.

Absorbance Spectrophotometry

4.1.4.1.1.

4.1.4.1.2.

4.1.4.1.3.

4.1.4.2.

Reflectance Spectrophotometry

4.1.4.2.1.

4.1.4.3.

Atomic Absorption Spectrophotometry

4.1.4.3.1.

4.1.4.3.2.

Flame Atomization

4.1.4.3.3.

Graphite Furnace

4.1.4.4.

4.1.4.4.1.

Principles of Fluorescence

4.1.4.4.2.

Excitation and Emission

4.1.4.4.3.

Applications in Clinical Chemistry

4.1.4.4.4.

Advantages and Limitations

4.1.4.5.

Chemiluminescence

4.1.4.5.1.

Principles and Applications

4.1.4.5.2.

Enhanced Chemiluminescence

4.1.4.6.

Turbidimetry and Nephelometry

4.1.4.6.1.

Principles and Differences

4.1.4.6.2.

Clinical Applications

4.1.4.6.3.

Particle Size Effects

4.2.

4.2.1.

Principles of Antigen-Antibody Reactions

4.2.1.1.

Antigen Structure

4.2.1.2.

Antibody Structure

4.2.1.3.

Affinity and Specificity

4.2.1.4.

Cross-Reactivity

4.2.1.5.

4.2.2.

Labels and Detection Systems

4.2.2.1.

4.2.2.1.1.

Horseradish Peroxidase

4.2.2.1.2.

Alkaline Phosphatase

4.2.2.1.3.

Beta-Galactosidase

4.2.2.2.

Radioactive Labels

4.2.2.2.1.

4.2.2.2.2.

4.2.2.3.

Fluorescent Labels

4.2.2.3.1.

4.2.2.3.2.

4.2.2.3.3.

4.2.2.4.

Chemiluminescent Labels

4.2.2.4.1.

Acridinium Esters

4.2.2.4.2.

4.2.3.

Competitive vs. Non-competitive Assays

4.2.3.1.

Mechanisms and Examples

4.2.3.2.

Advantages and Limitations

4.2.3.3.

Dose-Response Curves

4.2.4.

Homogeneous vs. Heterogeneous Assays

4.2.4.1.

Mechanisms and Examples

4.2.4.2.

Advantages and Limitations

4.2.4.3.

Separation Methods

4.2.5.

Common Immunoassay Formats

4.2.5.1.

Enzyme-Linked Immunosorbent Assay (ELISA)

4.2.5.1.1.

4.2.5.1.2.

4.2.5.1.3.

4.2.5.1.4.

Competitive ELISA

4.2.5.2.

Radioimmunoassay (RIA)

4.2.5.2.1.

4.2.5.2.2.

Safety Considerations

4.2.5.3.

Enzyme Multiplied Immunoassay Technique (EMIT)

4.2.5.3.1.

4.2.5.4.

Fluorescent Polarization Immunoassay (FPIA)

4.2.5.4.1.

4.2.5.5.

Chemiluminescent Immunoassays (CLIA)

4.2.5.5.1.

4.2.5.6.

Lateral Flow Assays

4.2.5.6.1.

4.2.5.6.2.

Point-of-Care Applications

4.3.

Electrophoresis

4.3.1.

Basic Principles

4.3.1.1.

Migration of Charged Particles

4.3.1.2.

Factors Affecting Separation

4.3.1.3.

4.3.1.4.

4.3.2.

4.3.2.1.

4.3.2.2.

Polyacrylamide Gel

4.3.2.3.

Cellulose Acetate

4.3.2.4.

Capillary Electrophoresis

4.3.2.4.1.

4.3.3.

Instrumentation

4.3.3.1.

4.3.3.1.1.

Voltage Control

4.3.3.1.2.

Current Monitoring

4.3.3.2.

Gel Casting and Loading

4.3.3.2.1.

Sample Application

4.3.3.2.2.

4.3.3.3.

Staining and Visualization

4.3.3.3.1.

4.3.3.3.2.

Nucleic Acid Stains

4.3.3.3.3.

4.3.4.

4.3.4.1.

Serum Protein Electrophoresis (SPEP)

4.3.4.1.1.

Normal Patterns

4.3.4.1.2.

Abnormal Patterns

4.3.4.2.

Hemoglobin Electrophoresis

4.3.4.2.1.

Normal Variants

4.3.4.2.2.

Abnormal Variants

4.3.4.3.

Isoenzyme Analysis

4.3.4.3.1.

4.3.4.3.2.

4.3.4.4.

Immunoelectrophoresis

4.3.4.4.1.

4.4.

4.4.1.

Basic Principles

4.4.1.1.

Partitioning Between Phases

4.4.1.2.

4.4.1.3.

4.4.1.4.

4.4.2.

Types of Chromatography

4.4.2.1.

Gas Chromatography (GC)

4.4.2.1.1.

Components and Operation

4.4.2.1.2.

Stationary Phases

4.4.2.1.3.

4.4.2.1.4.

Sample Preparation

4.4.2.2.

High-Performance Liquid Chromatography (HPLC)

4.4.2.2.1.

Components and Operation

4.4.2.2.2.

Stationary Phases

4.4.2.2.3.

4.4.2.2.4.

Sample Preparation

4.4.2.3.

Thin Layer Chromatography (TLC)

4.4.2.3.1.

4.4.2.4.

Ion Exchange Chromatography

4.4.2.4.1.

4.4.3.

Mass Spectrometry (MS)

4.4.3.1.

Principles of Mass Spectrometry

4.4.3.2.

Ionization Methods

4.4.3.2.1.

Electron Impact

4.4.3.2.2.

Chemical Ionization

4.4.3.2.3.

Electrospray Ionization

4.4.3.3.

4.4.3.3.1.

4.4.3.3.2.

4.4.3.3.3.

4.4.3.4.

4.4.3.4.1.

Electron Multiplier

4.4.3.4.2.

Photomultiplier

4.4.3.5.

4.4.3.5.1.

4.4.3.6.

4.4.3.6.1.

4.4.3.7.

Tandem MS (MS/MS)

4.4.3.7.1.

4.5.

Electrochemistry and Chemical Sensors

4.5.1.

4.5.1.1.

Principles of Potentiometry

4.5.1.2.

Reference Electrodes

4.5.1.3.

Ion-Selective Electrodes (ISE)

4.5.1.3.1.

Glass Electrodes

4.5.1.3.2.

Solid-State Electrodes

4.5.1.3.3.

Liquid Membrane Electrodes

4.5.1.3.4.

Applications in Electrolyte Measurement

4.5.2.

4.5.2.1.

Principles and Applications

4.5.2.2.

Glucose Sensors

4.5.2.3.

Oxygen Electrodes

4.5.3.

4.5.3.1.

Principles and Applications

4.5.3.2.

Chloride Analysis

4.5.4.

4.5.4.1.

Principles and Applications

4.5.5.

4.5.5.1.

Enzyme-Based Biosensors

4.5.5.2.

4.5.5.3.

4.5.5.4.

Clinical Applications

4.6.

Automation in the Clinical Laboratory

4.6.1.

Principles of Automation

4.6.1.1.

Workflow Integration

4.6.1.2.

Error Reduction

4.6.1.3.

Throughput Enhancement

4.6.1.4.

Cost Considerations

4.6.2.

Continuous Flow Analysis

4.6.2.1.

4.6.2.2.

4.6.2.3.

Flow Injection Analysis

4.6.3.

Discrete Analysis

4.6.3.1.

4.6.3.2.

4.6.3.3.

Batch Processing

4.6.4.

Modular Analyzers

4.6.4.1.

Types and Configurations

4.6.4.2.

Integration with Laboratory Information Systems

4.6.4.3.

Maintenance Requirements

4.6.5.

Point-of-Care Testing (POCT)

4.6.5.1.

Types of POCT Devices

4.6.5.2.

Quality Control in POCT

4.6.5.3.

Connectivity and Data Management

4.6.5.4.

Regulatory Considerations

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