HISTOCHEMISTRY, IMMUNOHISTOCHEMISTRY AND ULTRASTRUCTURAL ANALYSIS

Degree course: 
Corso di First cycle degree in Biomedical Laboratory techniques
Academic year when starting the degree: 
2022/2023
Year: 
2
Academic year in which the course will be held: 
2023/2024
Course type: 
Compulsory subjects, characteristic of the class
Seat of the course: 
Varese - Università degli Studi dell'Insubria
Credits: 
1
Period: 
First Semester
Standard lectures hours: 
14
Detail of lecture’s hours: 
Lesson (14 hours)
Requirements: 

The student, to access the course, must have knowledge and skills acquired in previous teachings, concerning human anatomy, physiology, general pathology, immunology, microbiology.

Final test will consist in an oral interview or a written exam that will focus on open questions or quiz on the topics covered in class.
The evaluation will take into consideration:
- detail, precision and significance in the theoretical contents acquired
- ability to present, argue and synthesize the leaned concepts with an appropriate language
- relevance and correctness
The personal re-elaboration of the learned contents and the ability to apply them to practical contexts will constitute a criterion of merit

Assessment: 
Voto Finale

The teaching activity aims to provide students with the basic tools to understand the techniques used in a Pathology Service.

Histochemical stainings for carbohydrates, nucleic acids, proteins, lipids, minerals.
Immunohistochemical methods and techniques.
Electron microscopy methods and techniques.

 HISTOCHEMISTRY
• Stainings
 General stainings in histochemistry
 Histochemical dyes: mechanisms of action
 General topographic stainings
• Carbohydrates
 Generalities
 Stainings:
o PAS
o Alcian blue
o Alcian-PAS
o Metachromatics methods
• Nucleic acids
 Generalities
 Stainings:
o Feulgen’s reaction
o Methyl-Piranine green
• Proteins
 Generalities
 Stainings:
o van Gieson’s trichromic stain
o Mallory’s trichromic stain
o Masson’s trichromic stain
o Orcein staining
o Weigert’s staining
• Lipids
 Generalities
 Stainings:
o Oil Red-O
o Sudan black B
o Blue Nilus
o OTAN
o Schiff’s method
o Schultz’s method
• Biogenic amines
 Generalities
 Stainings:
o Chromaffin reaction
o Masson-Fontana staining
o Grimelius’ method
• Pigments
 Generalities
 Stainings:
o Schmorl’s method
o Perls’ method
• Minerals
 von Kossa method for calcium
 Perls’ method for iron
 Rubeanic acid method for copper
• Enzymes
 Generalities
• Tessuto nervoso
 Generalities
 Stainings:
o Cajal’s method
o Weigert’s method
o PTAH
• Muscolar tissue
 Generalities
 Stainings:
o nitro-BT method
• Hemopoietic tissue
 Generalities
 Stainings:
o giemsa
o PAS
o Silver impregnation method for reticular fibres
o Perls
• Amiloidosis
 Generalities
 Congo red
• Bacteria, fungi and othes microbic organisms
 Bacteria
o Gram
o Ziehl-Nielsen
o Warthin-Starry
 Fungi
o PAS
o Grocott
 Viruses
 Protozoa
o Giemsa
 IMMUNOHISTOCHEMISTRY
• Definition
• Antigens and antibodies
• Monoclonal and policlonal atibodies
• Techniques and methods in immunohistochemistry
 immunofluorescence
 enzymatic immunohistochemistry
 immunogold
• Multiple antigens colocalizations
 Serial sections
 Multiple stains with eluition
 Multiple stains without eluition
• Controls in immunohistochemistry
• Merkers in immunohistochemistry
 ULTRASTRUCTURAL ANALYSIS
• Electron microscopy
 History
 Transmission electron microscopy
 Scanning electron microscopy
• Inclusion
 Sampling
 Fixation
 Epoxydic resins
 Hydrophylic resins
 Grids and support films
• Sectioning
 Semi-thin sections
 Ultra-thin sectioning
• Staining
 Lead salts, uranyl salts
Ultrastructural immunocytochemistry.

The teaching is proposed through lectures conducted by the teacher with the use of support materials (slides, example models, documents, bibliographic materials ...).
Students are required to systematically proceed with the independent study of the provided material, prepare the activities to be carried out in the classroom (if needed), and actively participate in the proposed discussions.
Students are always invited to contact the teacher in case of need for further information or clarification.

Prof. Facco is available for students. Please contact using the following e-mail: carla.facco@asst-settelaghi.it

Professors

FACCO CARLA