Oil immersion field Information and Courses from MediaLab, Inc.

The actual cell count is performed using the oil (100x) objective. This oil immersion field shows a representative counting field. Four granulocytes, a prorubricyte, and two rubricytes are completely visible here. 100 to 500 nucleated cells are generally counted,depending on the cellularity of the smear, and only cells completely visible in the field should be included in the count. The morphology of erythroid and myeloid precursor cells is described in detail in "Introduction to Hematopoiesis" and will not be repeated here.

Epithelial cells in large numbers within sputum smears means that the specimen is predominantly oral saliva, rather than true sputum from the lung. Epithelial cells in urine smears indicate that the sample has been contaminated by organisms found on the vulva or distal urethra. Bacteria found near or on epithelial cells are usually normal contaminating bacterial flora.White blood cells indicate inflammation and possible infection. The direct smear examination should focus within and around these cells.Red blood cells in a direct smear are not usually significant.Yeast may be present as normal flora in upper respiratory tract or genital tract. They may be significant if they predominate, or if budding yeast forms are seen.Hyphae are more likely to indicate the presence of fungal infection, but this determination requires correlation with clinical findings.Bacteria found in spinal fluid, blood, tissue and specimens from other sterile sites are always significant.Body fluids which are normally sterile must be examined carefully. If only one organism per oil immersion field is identified, then there are about 105 organisms per mL present in the sample! Bacteria observed in specimens from the throat, genital tract and other areas containing normal flora suggest infection only if their composition and type varies significantly from the norm.

In a study on the reporting of red blood cell morphology abnormalities conducted in Ontario, Canada (Hookey L, Dexter D, Lee DH, Laboratory Hematology 7:83-88, 2001), fewer than 50% of 33 participants used the same term to describe the quantitative frequency of peripheral blood abnormalities. Seven blood smears, each containing one of several abnormal erythrocytes-- schistocytes, teardrop cells, acanthocytes, and Howell-Jolly bodies--were evaluated by 32 participants. The participants were asked to document their evaluations from a list of quantitative terms. There was a heterogeneity in the use of terms "rare," "slight," "occasional," "few," "mild", "present," "moderate," "many," and "marked." Choices of terms were subjective without points of reference. Guidelines for establishing standardized qualitative estimations of abnormal erythrocytes in the peripheral smear are presented as follows: 1+ = 2 - 4/Oil Immersion Field (OIF) 2+ = 5 - 7/OIF 3+ = 8 - 10/OIF 4+ = >10/OIF. The terms "few," "moderate," "many," and "marked" may be substituted for the 1+ - 4+ grading system, but only when their specific points of reference are universally understood in tandem with the above guidelines. A comment should be triggered if any erythrocyte abnormalities are seen in numbers >3/OIF including, but not limited to, polychromasia, basophilic stippling, nucleated RBC's, and Howell-Jolly bodies. Rouleaux or RBC agglutination are important findings and must be documented.

The large cell in the center of this field is slightly blue/gray and is an example of a polychromatophilic red cell.Increased numbers of these cells, (approximately 2 or greater per oil immersion field,) indicate increased red cell output by the bone marrow. Polychromatophilic cells are larger and younger than mature red cells, and may be larger than 9 micron in diameter. Under normal conditions, these young red cells remain in the bone marrow one or two days before release into the bloodstream. However, when the bone marrow is stressed due to blood loss or other conditions, these cells are prematurely released into the blood, resulting in a blood smear with polychromasia. These red cells are often referred to as shift cells. If stained with a supravital stain, they would be identified as reticulocytes.