Histological Methods in Life Sciences

General description

The course focuses on the contemporary histological methods in biomedical research. These include histology, immunohistochemistry/immunofluorescence, FISH, and in vivo staining techniques. The course teaches how to efficiently collect and process fresh-frozen and formalin-fixed specimens, and how to conduct paraffin- and resin-embedding.
Its objective is to provide or improve practical skills of students in histological techniques, and empower their relevant application in biological and medical research.

Lectures
1. Organization and maintenance of histological laboratory

Why do we need histology? Development of histological techniques. Histological laboratory: general issues of location, organization and infrastructure. Equipment of the histological laboratory. Safety in the histological laboratory. Documentation and accounting. Features of procurement of equipment and reagents.

2. Harvesting, fixation and processing of animal tissues

Methods for collecting and fixing samples (grossing). Decalcification of samples. Methods of histoprocessing. Infiltration and embedding. Histoprocessing of unfixed material (fresh-frozen sectioning). Microtomy. 

3. Histological staining techniques

General principles of histological staining techniques. Basic techniques of histological staining. Hematoxylin: basic formulations. Hematoxylin-eosin staining: a routine method in histological practice. Staining methods for bacteria and fungi. Connective tissue staining. Methods of nervous tissue staining. Enzyme histochemistry.

 

4. Processing of hard tissues and implant materials

Bone tissue processing strategies. Histological processing of native (non-mineralised) bone. Grinding using the EXACT® system. Assessment of the bone-implant interface. Assessment of the implant surface. In vivo (vital) staining of the bone tissue. Decalcification. 
 

5. Immunohistochemistry 

General principles of immunohistochemical method. Production of specific antibodies. Antigen retrieval. Blocking of endogenous peroxidase. Blocking of nonspecific antibody binding. Visualization. Immunofluorescence technique.
 

6. In situ hybridization: FISH and CISH

History of the DNA hybridization method. Principle of the in situ hybridization method. Opportunities and challenges. In situ hybridization in the diagnosis of malignant tumors. Detection of ERBB2 (HER2/neu) amplification as a powerful prognostic tool in the treatment of breast cancer. Prospects for use.
 

7. Molecular genetics

DNA extraction. Polymerase chain reaction. Sequencing. Data analysis methods.
 

8. Data acquisition and image quantification techniques

The principle of operation of a light microscope. Modifications of the transmission light microscope.  The video demonstrates the operation of the AXIO Imager A2 microscope in light-optical and fluorescence modes, and provides an example of image processing in the ZEN program.

Final test
Рекомендована література
Підручники

•    Bancroft’s theory and practice of histological techniques, 2012. 7th ed. Elsevier. 
•    Lillie, R.D., Fulmer, H.M., 1976. Histopathologic technic and practical histochemistry, fourth ed. McGraw-Hill, New York.
•    Warford A., Nadège Presneau. Molecular Diagnostics. Oxford University Press, 2019

Онлайн-ресурси

•    Пошук гістологічних забарвлень онлайн: https://www.stainsfile.com/ 
•    Пошук імуногістохімічних і гістологічних методик онлайн https://www.ihcworld.com/ 

Level
Бакалавранти, магістранти, аспіранти
Lectures
8 Відеолекцій
Duration
16 годин
Language
Українська
Certificate
1 кредит ЄКТС
Lecturers

Associate professor of the Department of Pathological Anatomy, Forensic Medicine and Pathological Physiology at Dnipro State Medical University.