Biological Imaging: Microscopy Principles and Techniques
Academic Year 2026/2027 - Teacher: ROBERTA PECORAROExpected Learning Outcomes
Knowledge and understanding
- Knowledge of the physical and biological principles underlying light and electron microscopy
- Understanding of sample preparation protocols for light and electron microscopy.
- Knowledge of key immunohistochemistry and immunofluorescence techniques and their applications in biological research.
- Understanding of the principles of microscopic image acquisition and interpretation.
Course Structure
The course consists of lectures and practical laboratory activities. Should the course be delivered in a hybrid or remote format, necessary adjustments to the previously stated arrangements may be introduced to ensure the planned curriculum as outlined in the syllabus.
Required Prerequisites
Knowledge of the structural organization of the eukaryotic cell and the histo-morphological organization of animal tissues.
Attendance of Lessons
Attendance of the courses is mandatory. Attendance is understood to have been acquired if the student has attended at least 60% of the curricular hours provided for by the discipline.
Detailed Course Content
Principles of light microscopy: resolution, contrast, magnification, and illumination systems. Light microscopy techniques: bright-field, phase-contrast, and fluorescence. Principles of electron microscopy: structure and operation of the electron microscope. Transmission electron microscopy (TEM): sample preparation, fixation, embedding, ultramicrotomy, and contrasting. Scanning electron microscopy (SEM): sample preparation, metal coating, and image acquisition. Principles of immunohistochemistry: antigens, antibodies, specificity, and sensitivity. Immunolabeling techniques: enzymatic immunohistochemistry and immunofluorescence. Immunolocalization techniques in electron microscopy. Qualitative and quantitative analysis of microscopic images. Applications of microscopy and immunohistochemistry techniques in biological and biomedical research. Activities include lectures and practical laboratory sessions where students will carry out electron and light microscopy as well as immunohistochemistry protocols, acquiring skills directly applicable to the professional world and scientific research.
Textbook Information
Colombo - Olmo. Biologia: Cellula e Tessuti. 2° Ed. Edi-Ermes
Notes provided by professor
Course Planning
| Subjects | Text References | |
|---|---|---|
| 1 | Principles of light microscopy: resolution, contrast, magnification, and illumination systems. | |
| 2 | Light microscopy techniques: bright-field, phase-contrast, and fluorescence. | |
| 3 | Principles of electron microscopy: structure and operation of the electron microscope. Transmission electron microscopy (TEM): sample preparation, fixation, embedding, ultramicrotomy, and contrasting. | |
| 4 | Scanning electron microscopy (SEM): sample preparation, metal coating, and image acquisition. | |
| 5 | Principles of immunohistochemistry: antigens, antibodies, specificity, and sensitivity. | |
| 6 | Immunolabeling techniques: enzymatic immunohistochemistry and immunofluorescence. | |
| 7 | Immunolocalization techniques in electron microscopy. | |
| 8 | Qualitative and quantitative analysis of microscopic images. Applications of microscopy and immunohistochemistry techniques in biological and biomedical research. |