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Course includes study material and exam.

Description: Learn how to categorize different types of fixing agents; to elaborate on concepts in fixation and preservation of cellular elements for different types of analysis; and to utilize principles of tissue/cell preservation and fixation in order to effectively demonstrate specific components of cells and tissues. 

Start Date: Upon registration

Completion: Up to 52 weeks

Learning Outcomes:

• Describe basic structure and reactions of protein in relation to fixation
• Define purpose/function of fixation

o   Identify chemical structure of acetone, formaldehyde, ethanol, formic acid, picric acid, methanol, acetic acid, mercuric chloride, glutaraldehyde, osmium tetroxide, potassium dichromate, glyoxal

o   Describe features of fixing agents:

o   B-Plus™ (zinc based fixatives)

o   Bouin's, Hollande

o   Ethanol and alcohol solutions

o   Formaldehyde, NBF

o   Glutaraldehyde

o   Michel transport medium

o   Zenker, Helly, B-5 and mercury-based fixatives

o   Proprietary reagents

Preservcyt^TM

Cytolyt^TM

IBF^TM

-AZF^TM

• Define preservation and differentiate from fixation.
• Identify fixation/fixative artifacts
• Describe the effects and optimization of temperature, volume of fixative, osmolality, concentration, time of exposure and [H+] concentration on fixation of tissue
• Describe how the size of tissue affects fixation
• State the optimal thickness of tissue for paraffin processing for light microscopy
• State the optimal thickness of tissue for electron microscopy
• Differentiate between non-coagulant and coagulant fixative
• Describe the mode of action and general classification, advantages, disadvantages and safety precautions associated with:

o   Formaldehyde

o   Ethanol and alcohol solutions

o   Picric acid

o   Osmium tetroxide

o   Mercuric chloride

o   Zinc compounds

• Define

o   Tolerant fixative

o   Intolerant fixative

o   Simple fixative

o   Compound fixative

o   Coagulant fixative

o   Non-coagulant fixative

o   Additive fixative

o   Non-additive fixative

• Define autolysis: cytolysis, karyorrhexis, karyolysis
• Define putrefaction
• Describe conditions/analyses when/where fresh, unfixed tissue is required
• Define/Describe the use of post-fixation
• Discuss the theories regarding mechanisms of various types of fixatives
• Describe handling, disposal and storage of reagents used for fixatives

o   Use of personal protective equipment (PPE)

o   Exposure limits (OEL)

Author: Masaye Tanaka,  ART, BSc, MEd

Version Date: October 2016

PEP hours: 10

CPS credits: 0

*Note: PEP hours and/or CPS credits will only be awarded upon successful completion of Final Exam.