Development of image analysis methods for analyses of chromatographic and electrophoretic patterns from chemical and biochemical studies

Abstract

Owing to the rapid development of microelectronic, image analysis is now widely applied to the fields of sciences, industry and medical treatment. The aim of this research work is to apply image analysis technique and develop new method, such as intensity volume to study Chromatographic and Electrophoretic Patterns from chemical and biochemical studies. Thin layer chromatography (TLC) is the one of popular techniques in separation chemistry. However, quantitative analysis is regarded as the major weakness of TLC. An image analysis system which consists of an optical scanner and an IBM PC compatible as well as a software package, TLCQA, were developed. Besides, a new quantiation method based on the intensity volume of a spot in the chromatogram for quantitative analysis was developed. With this method, both qualitative and quantitative analyses can be conducted with high accuracy for the thin layer chromatographic technique. Nowadays, DNA analysis is routinely used in public and commercial forensic studies for investigating criminal cases, paternity disputes and others. An image system with the software, DNAFIN, was developed in this work to capture and process the images of silver stained gels from the DNA study automatically. Methods are developed to match the bands of standards and samples, sex identification and give an objective information to the operator for the judgment of stutter bands. The system definitely can assist the experts in DNA analysis. In DNA study, nucleic acid blotting and hybridization techniques are usually employed. Dot blotting is a widely applied in this study because of its fast and convenient. However, the quantitation of the intensity of hybridization in dot blotting always faces a great problem. A novel method was developed to quantify the dots and compare the difference between the corresponding dots in two different membranes. In this research, terminal differentiation of rat cardiac myocytes was selected as an example to test the performance of the method. Image analysis technique was also employed to study one of the most valued herbs in Chinese medicines (CMs), ginseng. TLC is one of the popular techniques to study the qualitative and quantitative analyses of CMs. Therefore, the method mentioned before was modified and used for studying the TLC patterns of the different types and products of ginsengs as well as American ginseng. As a result, a simple and accurate method was developed for quality control of ginseng products. Finally, a multimedia database, ChemBase, was developed for collecting, searching and re-organizing existing information of CMs. In addition, ChemBase provides both Chinese and English version with different searching methods. Besides the general information, chemical analysis information, such as thin layer chromatograms, were collected in ChemBase and they can be further analysed by image analysis method. Hopefully, a standard and useful chemical record of CMs can be set up.