Monocyte Information and Courses from MediaLab, Inc.

There are several diluents that may be used for CSF counts. Normal saline should be used to make dilutions for total cell counts. Diluting fluids for WBC counts include:crystal violet/acetic acidgentian violet/acetic acidtoluidine blue 0 and saponinThese fluids stain the white cells and lyse the red cells. The red cell count can be obtained by subtracting the white cell count from the total count. Low power (10x) may be used for the total count while the high power objective (40x) is suggested for the white cell count, especially if the white cells are to be differentiated into segs, lymphs and monocytes.

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.

Two monocytes are present in this field.

The arrow in this slide is pointing to a monocyte. The nucleus has an open chromatin pattern which gives it a spongy appearance. There is another monocyte in the lower right corner of the field. The other two cells could be classified as macrophages (histiocytes) because the nucleus is oval or kidney bean-shaped and the cytoplasm is very irregular. After circulating in the blood for one to three days, monocytes enter the tissues. The tissue form of the monocyte is called a macrophage or histiocyte.

Several types of cells are present in this slide from a patient who has suffered a cerebral hemorrhage. Lymphs, segmented neutrophils, one monocyte and mature red cells can be easily identified.

A 63 year old man was seen in the emergency room with the complaints of sudden onset of fever, chills, and abdominal pain, accompanied by mild diarrhea. The blood pressure was 140/84, the pulse rate 82/minute, and the body temperature 39.8C. A blood sample was drawn for a complete blood count, and a blood culture.A second blood culture was drawn from the opposite arm, with 10 ml of blood being placed into each an aerobic and an anaerobic bottle, following customary practice.The complete blood count revealed a hemoglobin of 15.8 mg/dl, a hematocrit of 45%, and a white blood count of 4.2/L. The neutrophils were 39%, lymphocytes 45%, monocytes 10%, eosinophils 4% and basophils 2%. The platelet count was 255/L. The patient was admitted to the hospital for further work-up and empiric antibiotic therapy.Within 24 hours after admission, the body temperature had decreased to 38.2C, although the mild diarrhea persisted.A stool toxin test for Clostridium difficile was negative and neither enteric pathogens nor Campylobacter species were recovered in stool culture after 24 hours incubation. Fecal neutrophils were not seen on direct examination. The anaerobic blood culture became positive 36 hours after inoculation.

Monocytes are phagocytes which remove injured and dead cells, cell fragments, microorganisms and insoluble particles from the blood and body tissues.Monocytes also secrete substances that affect the function of other cells, especially lymphocytes.They are produced in the bone marrow, and when mature are released into the peripheral blood. Although they do serve a phagocytic role in the blood, their main site of action is the body tissues.The half-life for monocytes in the peripheral blood is approximately 8 hours. Monocytes migrate into the tissues, often to sites of inflammation, where they serve their primary purpose.Here they transform into fixed or free macrophages, and continue their function as avid phagocytes.When activated, macrophages may enlarge and have enhanced metabolism.

The second group of cells are the leukocytes or white blood cells (WBC's). The leukocytes can be divided into two groups: mononuclear and granulocytic cells. Leukocytes are involved in various ways with the body's defense mechanisms. The cell shown in the upper image is a mononuclear cell, in this case a monocyte. The cell shown in the lower image is a granulocyte, in this case a neutrophil. These cells will be presented in much more detail later.

The nucleus may be round, kidney-bean shaped, folded, indented, or horseshoe, and may show "brain-like" convolutions.

The monocyte cytoplasm is opaque rather than transparent. One would be unable to read imaginary print placed behind the cell.

Because monocytes are extremely motile cells, blunt pseudopods may be seen. These should not be confused with the apparent cytoplasmic projections produced when large lymphocytes are indented by surrounding cells.

At times it can be very difficult to differentiate monocytes from large lymphocytes.Monocytes may be mistaken for large lymphs when their cytoplasm stains too lightly, when the characteristic granules are indistinct, or when the nucleus is rounded or only slightly indented.Sometimes a cell will have the nucleus of a lymphocyte and the cytoplasm of a monocyte, or some other confusing combination of characteristics.In order to properly identify the cell, it is necessary to weigh all of the characteristics together to determine which cell type it most resembles.Even then it is occasionally necessary to judge the cell on the basis of the company it keeps.For instance, if there are many monocytes, but few large lymphocytes around, the confusing cell is probably a mono.

Urine sediment may also contain white blood cells (WBCs). Most of the WBCs in urine are segmented neutrophils. Since it is possible that lymphocytes, monocytes, and/or eosinophils may be present, the cells in urine can be stained if it is necessary to differentiate them. The segmented neutrophil just above center of the image to the right shows a distinct nucleus. When viewing urinary sediment under the microscope, the fine focus adjustment must be used to identify white blood cells. White blood cells swell in dilute alkaline urine and the cytoplasmic granules exhibit brownian movement resulting in “glitter cells.” These cells lyse rapidly. “Glitter cells” are most easily seen when viewed under phase-contrast microscopy.

May-Hegglin bodies stain blue and appear to have a more definite shape than Dohle bodies. When examined under electron microscopy, they appear to be aggregates of thread-like structures in a crystal-like arrangement. May-Hegglin inclusions are RNA material believed to be derived from the endoplasmic reticulum. May-Hegglin bodies can be seen in monocytes and platelets as well as in all mature granulocytes. The platelets in May-Hegglin anomaly are very bizarre in appearance and thrombocytopenia is usually noted. When examining a slide that has bizarre platelets and blue-appearing bodies in the cytoplasm, thought should be given to the possibility of the May-Hegglin anomaly.

The neutrophils found in Chediak-Higashi can be differentiated from toxic granulation. In conditions causing toxic granulation, the granules are smaller and more numerous and only the neutrophils are affected. In Chediak-Higashi, eosinophils, basophils, lymphocytes and monocytes are affected. In eosinophils larger than normal eosinophilic granules may be seen, basophils may exhibit larger than normal basophilic granules, lymphocytes, large azurophilic granules. Larger pale granules/bodies may appear in monocytes.

Alder Reilly Anomaly is a rare autosomal recessive hereditary disorder in which the basic defect involves protein-carbohydrate complexes called mucopolysaccharides. The accumulation of partially degraded (broken down) protein-carbohydrate complexes within the lysosomes account for the larger than normal purple-staining granules seen in the granulocytes, monocytes and/or lymphocytes. The granules may occur in clusters, rather than diffusely, throughout the cytoplasm as in toxic granulation. These inclusions may be seen in the bone marrow more frequently than in peripheral blood. The physical characteristics associated with this disorder include gargoylism and dwarfism. The function of the cells involved is not affected. This morpholical change would be classified as pathological since the body is responding abnormally even though the function is not affected.

The following pages in this presentation includes a series of white blood cell abnormalities that may be identified in a peripheral blood smear. Many of the cases will simulate the practice of a peripheral smear review by a hematology morphologist. He/she must asses what responses in patient care may be triggered by the clinician attempting to interpret the reported findings on a peripheral smearObservations of white blood cell abnormalities in the peripheral blood smear should be reported so as to direct the physician to an immediate specific diagnosis, such as: (1) atypical lymphocytes suggesting infectious mononucleosis rather than leukemia, (2) toxic granules in neutrophils as in acute infections, or atypical granules suggesting a genetic disorder, (3) an unusual mix of cells, such as too many or too few neutrophils, monocytes, or other myeloid cells, and (4) the presence of giant platelets, myelocytes, or other cells suggesting a myelodysplastic syndrome.In summary, laboratory data should be presented to clinicians in a user friendly way to promote effective decision making. The design of the data base of information must be directed toward providing clinically helpful information clearly and quickly in order to facilitate appropriate action in terms of optimizing patient care outcomes.d

Large inclusions in leukocyte cytoplasm appear with Alder-Reilly syndrome. Inheritance patterns are not completely clear. The condition is characterized by larger than usual azurophilic and deeply violet staining granules clustered throughout the cytoplasm (even covering the nucleus)in all granulocytes. There are variations in which some lymphocytes and monocytes may be affected. These inclusions represent partially degraded mucopolysaccharides within lysosomes.Alder-Reilly bodies may be found independently of genetic mucopolysaccharidoses as an inherited anomaly (Jordan's anomaly). Cytoplasmic vacuoles of toxic origin are not present in Alder-Reilly cells. The background condition in Alder-Reilly syndrome is mucopolysaccharidosis with various types of bone and cartilage disorders, reported first in gargoylism, then in Hunter and Hurler syndromes. Accompanying conditions are hepatosplenomegaly, corneal opacities, and mental retardation. Reference: Brunning, Richard D. Morphologic Alterations in Nucleated Blood and Marrow Cells in Genetic Disorders. Human Pathol: 99-124, March, 1970

An 80 year old man was seen in the emergency room with sudden onset of right sided chest pain accentuated on inspiration. His cough was productive of yellow sputum, and he was short of breath.His temperature was 101.2F. A chest X-ray revealed right middle lobe pneumonia. His hemoglobin was 15.2 gm/dl, HCT 44%, and RBC 4.5 m/ml. The white blood count was 35,000/cuml, with 45% neutrophils, 20% bands, 5% lymphocytes, 3% eosinophils, 2% basophils, and 25% atypical monocytes as noted in the photograph.The atypical monocytes had abundant blue-grey cytoplasm with a few scattered vacuoles, which, in company with toxic neutrophils appeared to be a response to infection.The patient had a past history of tuberculosis which may account for the monocytosis.