Prussian blue Information and Courses from MediaLab, Inc.
These are the MediaLab courses that cover Prussian blue and links to relevant pages within the course.
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| Which one of the following statements about serum ferritin are true: | View Page |
| When a few small, purple inclusions are found in erythrocytes, they can be confirmed as containing iron by: | View Page |
| The cell indicated by the arrow on this Prussian blue stained smear is a siderocyte. | View Page |
| This Prussian Blue stain shows a sideroblast. | View Page |
| More on Pappenheimer bodies Pappenheimer bodies, while visible on a Wright's stained smear, should be Perls' Prussian blue stain, which is specific for iron. Pappenheimer bodies are seen in certain types of anemia characterized by an increase in the storage of iron, such as sideroblastic anemia and thallassemia. These inclusions are also seen in the peripheral blood following a splenectomy. In a healthy person with a normal spleen, Pappenheimer bodies are destroyed before the erythrocytes enter the peripheral circulation. | View Page |
| Match the following terms with the correct definition. | View Page |
| What is the reason for using Perl's Prussian blue stain? | View Page |
| Siderocytes This slide is stained with Perls' Prussian blue. The cell indicated by an arrow is a siderocyte. The inclusions (siderosomes) in the cell may or may not have been visible on Wright's stain. If they were visible on a Wright's stained smear, they would be designated as Pappenheimer bodies. | View Page |
| This Prussian blue stained smear shows: | View Page |
| Siderotic Granules To verify that red cell inclusions contain iron, it is necessary to use an iron stain, such as Perl's Prussian blue. The iron-containing granules are called siderotic granules. A mature erythrocyte containing siderotic granules is referred to as a siderocyte, while an immature (nucleated) erythrocyte containing siderotic granules is known as a sideroblast. A Pappenheimer body is a siderotic granule which is visible on Wright stain. All Pappenheimer bodies are Prussian blue positive, but not all siderotic granules are visible on Wright's stain as Pappenheimer bodies.
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| Siderotic Granules This slide has been stained with Perls' Prussian blue stain. The siderotic granules present in some of the red cells are stained blue-green, indicating its iron composition. These cells are called siderocytes. | View Page |
| Siderocytes and Ring Sideroblasts. 20 to 60% of red cell precursors seen in bone marrow slides normally contain siderotic iron granules visible with Prussian Blue stain. The presence of sideroblasts and siderocytes indicates that the red cell precursors have an ample supply of iron. When a red cell precursor contains too much iron, the siderotic granules form a ring around the nucleus and the resulting cells are referred to as ring sideroblasts. The ring sideroblast is an abnormal (pathological) form of sideroblast. | View Page |
| More on Erythrocyte inclusions The appearance, composition and associated physiology is specific for each type of inclusion. Identification and quantification of these inclusions is important because their presence may indicate an abnormality in the red cell system. Each of the inclusions listed above can be seen in more than one condition. There are erythrocyte inclusions specific to disorders which cannot be seen with either Wright-Giemsa stain or Perls' Prussian blue iron stain.
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| Storage Iron Storage forms normally comprise approximately 27% of total body iron. Stored iron provides a source of iron when physiologic demand is high, such as in blood loss, pregnancy, and periods of rapid growth. Storage compounds include ferritin and hemosiderin. Ferritin is a protein-bound, water-soluble, mobilizable storage compound and is the major source of stored iron. Hemosiderin is a water-insoluble form that is less readily available for use. When the amount of total body iron is relatively low, storage iron consists predominately of ferritin. When iron stores are increased, hemosiderin predominates. Unlike ferritin, hemosiderin stains with the Prussian blue stain (Perls reaction) and may be observed in tissues. | View Page |
| Definitive Tests for Iron Overload Measuring the amount of iron deposited in the liver is considered definitive for iron overload. This may be done by liver biopsy, computed tomography (CT), or magnetic resonance imaging (MRI). Demonstrating iron in parenchymal liver cells helps determine disease severity. Liver sections obtained by biopsy are stained with Perls Prussian blue which stains iron present in parenchymal cells. A photomicrograph of this reaction is shown.Although liver biopsy may not be necessary for diagnosing hereditary hemochromatosis (HH), it offers the advantage of detecting liver fibrosis if present. Molecular tests for mutations associated with HH are considered the gold standard of current HH testing. Liver biopsy is not needed for diagnosing all patients suspected of having HH, but may be ordered in some cases. | View Page |
| Match each of the following: | View Page |
| Fixation and Processing of the Biopsy Specimen The biopsy specimen is usually fixed in Zenkers’ acetic solution (5% glacial acetic acid; 95% Zenkers) for 6-18 hours or B-5 fixative for one to two hours. Excessive time in either fixative makes the tissue brittle, then briefly decalcified.
The tissue is processed together with other tissues, and is embedded in paraffin and cut at 4 µm and stained with hematoxylin and eosin (H&E) and Perls’ Prussian blue iron stain, and other special stains as indicated. | View Page |
| Perl's Prussian Blue Stain Perls' Prussian blue stain is used to detect the presence of iron which has not been incorporated into hemoglobin. Some unincorporated iron should be found in normal marrow. Perls' Prussian blue has been used for all of the slides in this exercise. With this stain, iron appears as a blue to blue/green staining material on all types of preparations used. | View Page |
| Pappenheimer bodies Pappenheimer bodies are iron-containing granules that aggregate with mitochondria and are deposited in RBC or normoblast cytoplasm. Small and irregular, they are found only in pathological states as thalassemia and sideroblastic anemias(upper image). Wright-Giemsa stain defines the cytoplasmic content (protein), but Prussian blue staining is necessary to define the iron content, the essence of the Pappenheimer body (lower image). Pappenheimer bodies lie typically in small clusters (upper image) and tend to locate at the periphery of the red cell cytoplasm. A cluster is typically smaller than a single Howell-Jolly body. | View Page |