lecture 4 Flashcards
why are reticulocytes called reticulocytes?
They are called reticulocytes because of a reticular (mesh-like) network of ribosomal RNA
what are the dark intracellular features found inside the reticulocytes?
vacuoles that contain remnants of organelles (ER, golgi, mitochondria) that are released from the cell as it matures
the ribosomal RNA of a reticulocyte is stained with which type of stain?
new methylene blue
what is the normal range of reticulocyte in men and women?
men: 0.5-1.5%
women: 0.5-2.5%
low reticulocyte count is seen in people with:
cirrhosis, folic acid deficiency, and bone marrow failure
high reticulocyte count is seen in people with:
bone marrow is responding to the need for increased RBC production (person who has recently donated blood or responding to treatment for anemia)
what is the procedure for reticulocyte count?
- put a drop of a supravital stain BCB (Brilliant Cresyl blue) on a slide and spread it
- slide will become blue
- air dry for 1 min
- put a drop of a supravital stain BCB (Brilliant Cresyl blue) on a slide and spread it
- Using a capillary tube, place a drop of anticoagulated blood on each side of the slide
- cover it with a cover slip (push a little on the cover slip + air dry for 10 mins)
- Using a capillary tube, place a drop of anticoagulated blood on each side of the slide
- using low-power objective (10x), find an area in the thin portion of the smear in which the red cells are evenly distributed and are not touching each other
- Carefully change to oil immersion (100x) and further locate an area in which there are approx 200 RBC per oil immersion field
- Select 5 fields.
- same is done for the second side of the slide
- using low-power objective (10x), find an area in the thin portion of the smear in which the red cells are evenly distributed and are not touching each other
- Count reticulocytes in 5 fields.
- Record the number of retics seen
- calculate the % of reticulocytes using formula
- Count reticulocytes in 5 fields.
what is the formula to calculate the % reticulocytes?
% Reticulocytes= number of reticulocytes counted in 1000 red cells /10
what are the stages of erythropoesis?
- myeloid stem cell
- proerythroblast = pronormoblast
- basophilic (early) erythroblast
- polychromatic (intermediate) erythroblast
- orthochromatic (late) erythroblast
- reticulocyte (polychromatic reticulocyte)
- erythrocyte
what are the major changes during the process of development of the RBC(s)?
- size of cell becomes smaller
- size of nucleus becomes smaller
- chromatin becomes denser
- basophilia gradually decreases until in the mature RBC there is no RNA.
proerythroblast key points
- first recognizable cell
- large cell
- large nucleus
- contains very fine-chromatin
- few nucleoli (1-4)
- cytoplasm is abundant + intensely basophilic (due to presence of a large amount of RNA)
- appears as a band around the nucleus
- can either undergo mitosis to make another proeythroblast or can mature to give the Basophilic/early erythroblast
Early erythroblast key points
- becomes smaller in size
- nucleus becomes smaller
- chromatin pattern becomes coarser (condensed)
- no more nucleoli
- cytoplasm is still intensely basophilic but less than
proerythroblast - can also divide to give another early erythroblast
or can mature to a polychromatophilic (intermediate) erythroblast.
Intermediate erythroblast key points
- cell becomes smaller
- nucleus becomes smaller
- chromatin more condensed
- cytoplasm is less basophilic and appears grayish to pinkish
- Hb starts to add up (production of Hb)
- can divide or mature to an orthochromic (late) erythroblast.
Late erythroblast key points
- cell smaller
- smaller nucleus
- nucleus is completely condensed (pyknotic nucleus)
- cytoplasm becomes more abundant + more pinkish
- no more able of further mitosis (because the nucleus is so condensed that the DNA material cannot replicate)
- nucleus is extruded, taken by phagocytes of the B.M
- cell becomes anucleated
what is the major modification in a basophilic erythroblast?
no more nucleoli