Wright-giemsa Information and Courses from MediaLab, Inc.
These are the MediaLab courses that cover Wright-giemsa and links to relevant pages within the course.
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| Stained Cytospin Preparations of CSF All white cells present in a cerebrospinal fluid must be identified.
If more than 10 cells/mm3 are present or there is difficulty identifying the few cells that are present, make a cytospin, a filtration, or a sedimentation preparation, stain with Wright-Giemsa, and perform differential count.
Cytospins made with a cytocentrifuge are preferred since they are easiest to make and interpret, but filtration and sedimentation methods can also be used to prepare a slide for subsequent staining.
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| Cytospin Technique In the cytospin procedure, use a high speed centrifuge to concentrate the cells on a slide in a uniform monolayer 6 mm in diameter. The monolayer distribution enhances the morphological appearance of the cells present.Allow the slides to dry in air for several minutes and then stain them with Wright-Giemsa stain. Cytospin slides may be placed in an automatic stainer, such as Hema-Tek, or stained manually.Perform a 100 or 200 cell differential and record the number of neutrophils, eosinophils, basophils, lymphocytes, monocytes, macrophages, and blasts cells.Pathologists must review any slide which has tumor cells, unidentified cells, or immature stages of cells, such as blasts.Since criteria for review may vary from one laboratory to another, be sure to check the requirements in your laboratory before reporting the differential. | View Page |
| Which of the following statements are true regarding spinal fluid differentials? | View Page |
| Which of the following is a routine staining technique used in the clinical laboratory: | View Page |
| Which of the following stains is not routinely used when examining peripheral blood smears ? | 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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| What is Erythrocyte inclusion? Erythrocyte inclusion is a term used to describe structures that may be present in red cells. The inclusions discussed in this course are: reticulocyte - Wright-Giemsa stain (polychromasia)basophilic stipplingHowell-Jolly bodiesPappenheimer bodies (Wright-Giemsa stain)siderotic granules (iron stain)Cabot rings | View Page |
| Biopsy Touch Preparation A touch prep is made by holding the biopsy plug with a forceps and touching the plug to one or more clean slides in several places. Imprints of the marrow remain on the slide. The slides are quickly air dried, fixed with methanol and stained with Wright-Giemsa or other cytochemical stains. Morphologic details of the cells are preserved with this type of preparation. | View Page |
| Romanowsky Stain "Romanowsky stain" is a general term which can include several specific stains or stain combinations. Wright-Giemsa is a Romanowsky type stain combination frequently used to stain bone marrow smears. | View Page |
| Examination of Wright-Giemsa Stained Bone Marrow Examination of Wright-Giemsa stained bone marrow preparation involves examination under low power (10X objective) high power (40-50X objective )and oil immersion (100X objective). Low power examination: Assess quality of smear, assess number of megakaryocytes.Assess myeloid to erythroid ratio.Evaluate morphology and do differential count. | View Page |
| Appearance of Cytoplasm Their cytoplasm is usually colorless or light blue. However, the color is usually masked by the large granules that are present. These granules take up the acid components of Wright-Giemsa stain, and are therefore reddish-orange. | View Page |
| Appearance of Cytoplasm The color of the cytoplasm is light pink to colorless, but as with eosinophils, the specific basophilic granules are prominent.These granules are large and, because they take up the basic components of Wright-Giemsa stain, are purplish-black. | 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 |
| Reticulocyte identification Reticulocytes are red blood cells prematurely released from the bone marrow. On a Wright-Giemsa stained blood smear, they appear as polychromatic macrocytes. Their presence in the peripheral blood may suggest hemolysis or bleeding. Their presence is expressed as a percentage of the red cell count: newly born= 3-7%; up to one week of age=1-3%; >one week =0.3-1.8%. Automated or manual methods may be used to enumerate reticulocytes. In clinical context, retics must be separated from debris, precipated stain, Pappenheimer bodies, Howell-Jolly bodies, and Heinz bodies. | View Page |
| Heinz body formation Heinz bodies are 1-3 um particles of denatured hemoglobin settling eccentrically, usually close to the red cell membrane. They are found in erythrocytes in unstable hemoglobin disorders, acute drug induced hemolysis, and following splenectomy. Their formation may be exaggerated by in-vitro incubation of a fresh blood sample with phenylhydrazine. Heinz bodies, as pictured here, are identified using a supra-vital stain, such as new methylene blue or cresyl violet. Bite cells, visible with Wright-Giemsa staining, are visual reminders that the spleen is functional and has pitted the aberrant chunk of hemoglobin from the circulating erythrocyte. | View Page |
| Hemolytic disease of the newborn Jaundice was recognized in a day-old infant. Notice particularly the size variation (anisocytosis) of the erythrocytes on the infant's peripheral smear. What does this observation mean? Does it provide immediate information that might serve as guidance in expediting diagnosis and treatment? Note that normal-sized red blood cells, microcytes, microspherocytes, macrocytes, and nucleated red blood cells are all present. Red cell variations are expected findings in healthy neonates, but the variations here are exaggerated. Hyposplenic functional features may appear, including acanthocytes, spherocytes, and possibly Howell-Jolly bodies, especially if hemolysis is particularly vigorous. A high (3-7%) reticulocyte count is not unusual during the first three or four days after birth, however, the marrow in this jaundiced infant is proliferating vigorously in response to hemolysis. A call for more red cells is urgent. Immature red cells (in the form of nucleated red cells) and red cells with stippling of RNA (basophilic stippling) are readily identified. Red cell maturation sequence has not been totally processed in the marrow nor is all residual red cell debris removed by the spleen. In the lower photograph are reticulocytes stained by supravital stain (new methylene blue). Basophilic stippling (specks of RNA) stains with both supravital stains and with routine Wright-Giemsa stain. | View Page |
| Staining Eosinophils Patients with hypersensitivity reactions, sometimes as a result of medications, may have eosinophils in their urine. Hansel's stain, which is specific for eosinophils, or Wright-Giemsa stain may be used to distinguish these cells from neutrophils. | View Page |