Lab-on-a-chip Technics in Laboratory Diagnostics

Data

Official data in SubjectManager for the following academic year: 2024-2025

Topic

Microchip capillary electrophoresis is an interesting approach for an increasing number of analytical problems in environmental applications and health science. The determination of size, quality and concentration of biomolecules such as DNA, RNA and proteins is one of the fundamental steps in life science research. Traditional methods for this type of analysis, such as gel electrophoresis or capillary electrophoresis, can now be complemented by an analytical technique, Lab-on-a-Chip technology. This technology enables downscaling and integration of several experimental steps (injection, labeling, dilution, separation, detection) into one process, in combination with automated data analysis. Lab-on-a-Chip technology has several advantages compared with conventional techniques, such as minimal sample requirement, rapid analysis times, ease-of-use, minimized exposure to hazardous materials and reduced waste generation.

The course deals with the theoretical background and application of the microfluidic methods, such as electrophoresis for detection of proteins, endotoxins, DNA, RNA and flow cytometry for detection of cells (bacteria, fungi, cancer cells, etc.).

Lectures

• 1. The principle and practice of capillary zone electrophoresis, theory of electrophoretic migration - Makszin Lilla
• 2. The principle and practice of gel electrophoresis, polyacrylamide gel electrophoresis (2-D PAGE, SDS-PAGE) methods - Makszin Lilla
• 3. The principle and practice of microchip electrophoresis - Makszin Lilla
• 4. The principle and practice of flow cytometry on microchip - Makszin Lilla
• 5. The structure and function of microchip electrophoresis device (Agilent Bioanalyzer 2100) (different types of detectors, injection methods) - Makszin Lilla
• 6. Preparation of microfluidic chips, properties and advantages - Makszin Lilla
• 7. Laboratory diagnostic applications of microfluidic chips 1. (electrophoretic separation of proteins, DNA, RNA) - Makszin Lilla
• 8. Laboratory diagnostic applications of microfluidic chips 2: (flow cytometry analysis of bacteria, fungi, cancer cells) - Makszin Lilla
• 9. Qualitative evaluation of „lab-on-a-chip” methods by help of Agilent 2100 Expert software - Makszin Lilla
• 10. Quantitative evaluation of „lab-on-a-chip” methods by help of Agilent 2100 Expert software - Makszin Lilla

Practices

• 1. Preparation of samples (serum proteins, bacterial proteins, endotoxins, etc.) - Makszin Lilla
• 2. Preparation of samples; covalent labeling with fluorescent dye - Makszin Lilla
• 3. Preparation of samples; denaturation - Makszin Lilla
• 4. Microchip electrophoretic measurement, separation with Agilent 2100 Bioanalyzer - Makszin Lilla
• 5. Qualitative analysis of samples - Makszin Lilla
• 6. Quantitative analysis of samples, evaluation by help of 2100 Expert software - Makszin Lilla
• 7. Preparation of cell samples (bacteria, fungi, HeLa cells, etc.) - Makszin Lilla
• 8. Purification of cells - Makszin Lilla
• 9. Preparation of samples; labeling with fluorescent dyes - Makszin Lilla
• 10. Purification of labeled cells - Makszin Lilla
• 11. Microfluidic measurement, cell counting and evaluation with Agilent 2100 Bioanalyzer - Makszin Lilla
• 12. Final evaluation, mark - Makszin Lilla

Seminars

Reading material

Obligatory literature

Literature developed by the Department

It is going to be available on the website and in Neptun.

Notes

Recommended literature

Instrumental analysis lecture and practise; microchip electrophoresis

Ann Van Schepdael: Microchip Capillary Electrophoresis Protocols, Springer, 2015

G2938-90062 Agilent Protein 80 Reagent Kit GuideUser Manual 9/19/2006

G2938-90310 Agilent (2008) Agilent High Sensitivity Protein 250 Kit Guide

G2946-90004 Agilent 2100 Bioanalyzer 2100 Expert User's Guide User Manual 6/21/2005

Conditions for acceptance of the semester

Active participation and written test.

Mid-term exams

-

Making up for missed classes

By personal consultation.

Exam topics/questions

The required information for the test will be explained in the course.

Examiners

Instructor / tutor of practices and seminars

• Makszin Lilla