Kimberley Pyneandy (1021635) 28th September, 17

Histotechnology (DMT 3105) Lab #2

Reaction of Fixatives with Carbohydrates

The purpose of fixation is to preserve tissue structure with minimal alteration during
dehydration, embedding, cutting, staining, and viewing in the electron microscope.
(Hunter, Maloney, & Bendayan, 1993)
There are many effects on carbohydrates from the reaction of fixatives. It could be
thought of as a preservation of carbohydrates since fixatives do not readily impact it. In
glycogen, a polysaccharide, it has low molecular weight and is more soluble in aqueous
media but not easy to fix. (Hans, Anderson, Hasselager, Moller, & Prento, 1991)
Artifacts are associated with poor fixing of glycogen, e.g., streaming artifact, which is in
the diffusion pathway of the fixative through the tissue. This streaming artifact is very
prominent when using fixatives with ethanol at room temperature. To combat the low
molecular weight of glycogen, it can be coated with nitrocellulose before staining. (Hans,
Anderson, Hasselager, Moller, & Prento, 1991)
Most formaldehyde fixatives work best for glycogen with higher molecular weights
(liver). Bouins fixative and Gendres fluid can be used to get good preservation of the
samples along with good results. However, the streaming artifact is produced when using
the Bouins fixative. Also freeze drying, and the use of fixation with formaldehyde gas
can be used since it showcases good results. (Hans, Anderson, Hasselager, Moller, &
Prento, 1991)
Being mainly proteins, Glycoproteins can be fixed using many good protein fixatives.
The solubility in water and to some extent in 70% ethanol is great for proteoglycans,
which causes the precipitation in 100% ethanol but still partially soluble in water. (Hans,
Anderson, Hasselager, Moller, & Prento, 1991)
The sensitivity of the test choice used for the demonstration of a particular carbohydrate
is a limiting factor and dictates the degree of efficiency. (Hunter, Maloney, & Bendayan,
1993)
The reaction of primary fixatives with carbohydrates using various fixatives, for example:
Aldehydes e.g. formaldehyde and glutaraldehyde preserves glycogen without being fixed,
chromium trioxide oxidizes glycol groups to aldehydes, ethanol and picric acid
precipitates glycogen without fixing the specimen. Potassium dichromate, mercuric
chloride, osmium tetroxide and acetic acid has no effect on carbohydrates. (Hans,
Anderson, Hasselager, Moller, & Prento, 1991)

Bibliography
Hans, L., Anderson, A., Hasselager, E., Moller, M., & Prento, B. (1991). Theory and
Strategy in Histochemistry: A Guide to the Selction and Understanding of Techniques.
Springer-Verlag.
Hunter, E., Maloney, P., & Bendayan, M. (1993). Practical Electron Microscopy: A
Beginner's Illustrated Guide (2nd Edition ed.). Cambridge University Press .
McManus, J., & Mowry, R. M. (1958). Effects of Fixation on Carbohydrates
Histochemistry. University of Alabama Medical Center, Department of Pathology ,
Birmingham.