Reticulocytes are immature red blood cells, typically composing about 1% of the red cells in the human body. Reticulocytes develop and mature in the red bone marrow and then circulate for about a day in the blood stream before developing into mature red blood cells. Like mature red blood cells, reticulocytes do not have a cell nucleus. They are called reticulocytes because of a reticular (mesh-like) network of ribosomal RNA that becomes visible under a microscope with certain stains such as new methylene blue.
To accurately measure reticulocyte counts, automated counters that use lasers mark cell samples with fluorescent dye that marks RNA and DNA (such as thiazole orange or polymethine). This distinguishes reticulocytes as the middle ground of dye response to laser light, between red blood cells (which have neither RNA nor DNA) and lymphocytes (which have a large amount of DNA, unlike reticulocytes).
The specimen requirement for a reticulocyte count is EDTA anti-coagulated whole blood (lavender-top bottle if using the Vacutainer, Vacuette or Monoject systems; red-top if using the S-Monovette system).
Reticulocytes appear slightly bluer than other red cells when looked at with the normal Romanowsky stain. Reticulocytes are also slightly larger, which can be picked up as a high MCV (mean corpuscular volume) with a full blood count done by a trained medical scientist, who has specialized in hematology, or a machine.
Flowcytometry for mouse reticulocytes: One can use a cell-permeable thiazole orange dye (see above) to stain for reticulocytes' residual RNA in conjunction with DRAQ5 DNA-only dye (reticulocytes have no DNA and are, thus, DRAQ5-negative) and Ter119 (glycophorin-A) that is a marker of erythroid lineage. (Thiazole orange dye binds to nucleic acids of both DNA and RNA).
The normal range of values for reticulocytes in the blood depends on the clinical situation and the lab but is usually 0.5% to 1.5%. However, if a person has anemia, the reticulocyte percentage should be higher than "normal" if the bone marrow's ability to produce new blood cells remains intact. Thus, calculating the reticulocyte production index is an important step in understanding whether or not the reticulocyte count is appropriate to the situation. This is often a more important question than whether the percentage is in the normal range; for instance, if someone is anemic but has a reticulocyte percentage of only 1%, the bone marrow is likely not producing new blood cells at a rate that will correct the anemia.
The number of reticulocytes is a good indicator of bone marrow activity because it represents recent production. This means that the reticulocyte count and the reticulocyte production index that can be calculated from it can be used to determine whether a production problem is contributing to the anemia and can also be used to monitor the progress of treatment for anemia.
When there is an increased production of red blood cells to overcome chronic or severe loss of mature red blood cells, such as in a haemolytic anemia, people often have a markedly high number and percentage of reticulocytes. A very high number of reticulocytes in the blood can be described as reticulocytosis.
Abnormally low numbers of reticulocytes can be attributed to chemotherapy, aplastic anemia, pernicious anemia, bone marrow malignancies, problems of erythropoietin production, various vitamin or mineral deficiencies (B9, B12, iron), disease states (anemia of chronic disease) and other causes of anemia due to poor RBC production.
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