Fixatives for Electron Microscopes
Fixation is a process in which tissue sections collected from dead or living organisms are preserved as close as possible to their original state when they were alive, to prepare microscopic specimens for biological or pathological observation. The purposes are as follows:
- To stabilize proteins, the major components of tissue sections, as quickly as possible in order to arrest autolysis
- To minimize the alteration and deformation of tissue sections caused by chemicals, heat, etc. in the subsequent specimen preparation process
While many fixation methods are currently available, it is necessary to select the most appropriate method depending on the microscope used and the target structure or substance.
Fixative solutions for electron microscopes
| Fixative solution | Concentration used | Feature | Defect | Target tissue |
|---|---|---|---|---|
| Osmic acid | 1～2 ％ | -Washing with water and dehydration are completed quickly. -Only slight deformation occurs during fixation. -Embedding agents penetrate easily, and various resins can be used. -Phospholipids are effectively fixed. |
-Fixative solutions penetrate poorly. -50% of proteins and saturated fatty acids are lost. |
-Intracellular ultrastructure -Unsaturated fatty acids -DNA particles |
| Potassium permanganate | 0.6～3 ％ | Fixative solutions and embedding agents penetrate moderately well. | Major deformation occurs during fixation. Mucopolysaccharides, proteins, and saturated fatty acids/phospholipids are partially lost. No ribosomes are fixed. |
-Intracellular membrane structure -Glycogen granules -Plant tissues |
| Glutaraldehyde | 1.5～4 ％ | Intracellular structures and glycogen granules exhibit negative staining, and nuclei and intracellular matrix exhibit positive staining. | Washing with water and dehydration takes time. Embedding agents (other than methacrylic resins) penetrate poorly. Lipids are not sufficiently fixed. Low-concentration proteins are not fixed. |
Polysaccharides |
| Tannic acid-aldehyde fixation | 1 ％, 2 ％ | Soluble proteins (functional proteins such as hormones and enzymes) and polypeptides are effectively fixed by mixing with aldehyde fixative solutions. | - | -Soluble proteins -Polypeptides |
| Neutral buffered formalin | 4 ％ | Soluble proteins (functional proteins such as hormones and enzymes) and polypeptides are effectively fixed. | - | -Soluble proteins -Polypeptides |
| Paraformaldehyde | 4 ％ | Glycoproteins are fixed, antigenicity is retained, and tissue penetration is rapid. | Fixation is weak. | Protein antigens |
| Periodate-lysine-paraformaldehyde (PLP) | 2％ | Glycoproteins and sugars are effectively fixed. Antigenicity is retained. |
- | Glycoprotein antigens |
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Fixative Solutions for Electron Microscopes
Fixative Solutions for Electron Microscopes (Ampule type)
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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