Methods in cytochemistry

A.Y. 2021/2022
Overall hours
Learning objectives
The aim of the course is to provide students with the basic principles of cytochemistry and histochemistry. The preparation techniques for sample analysis used in cytochemistry, histochemistry, immunocytochemistry and in situ hybridization will be illustrated. Basic knowledge will also be provided for optical and electronic visualization of biological interest macromolecules. Theoretical lessons will be associated with seminars / workshops relating to examples of application of these methods to the study of cellular biology and instrument observation tools.
Expected learning outcomes
At the end of the course, the student will have acquired basic theoretical knowledge related to:
- preparation techniques of histological samples and their cyto-histochemical staining;
- main techniques of immunohistochemistry and in situ hybridization;
- main methods of in situ observation of biological macromolecules in cells and tissues.
Course syllabus and organization

Single session

Lesson period
Second semester
More specific information on the delivery modes of training activities for academic year 2021/22 will be provided over the coming months, based on the evolution of the public health situation.
Course syllabus
The instruments of microscopic observation. Principles of geometrical optics. Image formation. Magnifying lenses. Compound microscope. Object lighting.
Beyond geometrical optics: wave nature of light, interference and diffraction phenomena. Resolving power of an optical device.
"Special" techniques in optical microscopy: brightfield, darkfield, phase contrast, and polarized light. Fluorescence microscopy. Fluorescence spectroscopy. Fluorochromes. Applications of fluorescence microscopy (FRAP, FLIP, FRET, BRET). Principles and instruments of the confocal microscopy. Super-resolution microscopy. Electronic microscopy.
Biological sample preparation techniques in optical microscopy: sampling procedures, physical and chemical methods of fixation, inclusion in paraffin and plastic resins, sectioning and relative instruments. Section staining for observation optical microscope.
Dyes: structure and chemical classification. Chromophores and auxochromes. Metachromasia. Histomorphological stains. Cytochemical stains. Immunohistochemical stains. In situ hybridization. Biological sample preparation techniques for electronic microscopy analysis.
Flow cytometry: basic principles, technical equipments, sample preparation, data management and analysis. Applications of flow cytometry in the basic and applied research. Basic principles of CyTOF mass cytometry. Applications of CyTOF mass cytometry in the basic and applied research.
Prerequisites for admission
Basic knowledge of cytology, histology, and biological chemistry.
Teaching methods
The teaching course will consist of classroom lessons and seminars deepening the topics covered in the Course. The lessons will be comprised of PowerPoint slides provided by the teacher and supplemented, when needed, by chalkboard explanations. The students will also be provided with copies of recent articles published on international journals inherent to the covered topics. These articles will also be available in pdf format on the teacher web site.
Attendance at the teaching course is highly recommended.
Teaching Resources
A.G.E. Pearse, Istochimica. Teoria e pratica, Piccin.
Assessment methods and Criteria
The final exam will consist of a written test with open-ended questions on the covered topics. The time to complete the test is 2 hours.
BIO/16 - HUMAN ANATOMY - University credits: 6
Lessons: 36 hours
Lessons (seminars): 12 hours
Educational website(s)