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List 5 different specific indications for a bone marrow biopsy.
- a diagnostic aid
- an unexplained increase in blood cells
- an unexplained decrease in blood cells
- when we see abnormal cells in peripheral blood
- to look for mast cells if MCT
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Do we use anesthesia to collect bone marrow? Why or why not?
- yes, local or general
- for aseptic technique
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What places on a small animal can we collect bone marrow?
- iliac crest
- trochanteric fossa
- proximal humerus
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What places on a large animal can we collect bone marrow?
posterior rib or iliac crest
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How do we collect the sample?
- place needle in lumen of the bone
- we want bone spicules
- use an anticoagulant
- either make 2 - 5 slides and stain one or place part in 10% formalin to send off for biopsy
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Describe the cytologic findings of a normal marrow sample.
- cells appear normal: size, N:C ratio, staining
- M:E ratio is within normal limits
- mature cells > immature cells
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List in order the megakaryocyte series.
- megakaryoblast
- promegakaryocyte
- megakaryocyte
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List in order the erythroid series.
- rubriblast
- prorubricyte
- rubricyte
- metarubricyte
- reticulocyte
- mature erythrocyte
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List in order the granulocytic series.
- myeloblast
- progranulocyte
- myelocyte
- metamyelocyte
- band
- mature granulocyte
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How do the cells of the erythrocyte series differ in appearance from those of the granulocyte series?
- in the erythrocyte series, the cells and nucleus are round and have basophilic cytoplasm
- in the granulocyte series, the nucleus goes from round to kidney shaped and the cytoplasm is pale blue
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Define plasma cell.
comes from lymphocytes and produce antibodies
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Define osteoclast.
breaks down bone.
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Define promonocyte.
Second stage in monocytopoiesis
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Define osteoblast.
makes bone
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Define Mott cell.
plasma cell full of russell bodies
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What is the M:E ratio and what does it tell the VMD? List 2 causes of a high ratio. List 2 causes of a low ratio.
- myeloid vs. erythroid
- high ratio: greater than 3...myeloid hyperplasia or erythroid hypoplasia
- low ratio: below 0.5...myeloid hypoplasia or erythroid hyperplasia
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Name 3 myeloproliferative disorders that affect the bone marrow.
- multiple myelonma
- lymphoma/small cell lymphoma
- mast cell tumor
- leukemia
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Where are blood cells produced?
in the medullary cavity of long bones
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What are the 5 common stem cells produced in the bone marrow?
- erythropoiesis
- granulopoiesis
- monocytopoiesis
- lymphopoiesis
- thrombocytopoiesis
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What does a bone marrow needle look like?
- large hub
- stainless steel
- 11 - 16 gauge
- short bevel
- stylet
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What supplies do we need to perform a bone marrow aspirate?
- #10 blade (to nick skin)
- 2 - 3 (12 ml syringes)
- bone marrow needle with stylet
- local or general anesthesia
- about 20 clean slides
- EDTA
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Describe how to aspirate a bone marrow sample.
- attach syringe to needle
- apply negative pressure - quickly
- release as soon as you see red stuff
- repeat until you see material in the hub and then stop
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How do we evaluate a bone marrow slide?
- scan on low then on high
- evaluate cells under oil
- identify/classify 500 cells
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What are the three different categories we should sort our 500 cells into?
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Other than counting cells, what is the other thing we do when we are evaluating a bone marrow slide?
calculate M:E ratio
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What kind of stain do we use for bone marrow slides?
diff quik
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Describe what the cells look like in the red cell series.
- round cells
- round nuclei
- narrow rim of bright blue cytoplasm
- get smaller as they mature
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What do the cells in the red cell series look similar to?
lymphocytes
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As the RBC matures, how does the cytoplasm change?
the cytoplasms size, shape, and color changes
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As the RBC matures, how does the nucleus change?
shape, color, and size
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What is pyknosis?
- dead nucleus
- whats in a nucleated red blood cell
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As the RBC matures, how does the chromatin change?
chromatin goes from vesicular to clumpy.
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What does a vesicular chromatin look like?
lacey
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Describe the look of a rubriblast.
- large cell
- narrow rim of bright blue cytoplasm
- round nucleus
- 1 - 2 nucleoli in nucleus
- lacey chromatin
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What is the nucleoli involved in?
protein synthesis
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If a cell has characteristics of 2 different stages of the cell, what stage do we classify it as?
the more mature stage
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Describe a prorubricyte.
same as a rubriblast EXCEPT no nucleolus present and has a perinuclear clear zone
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Describe a rubricyte.
- smaller cell
- round nucleus
- clumpy, coarse chromatin
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Describe a metarubricyte.
- even smaller than a rubricyte
- cytoplasm blue to pinkish
- nucleus small and dark and eccentric
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What stage of the red cell series does mitosis stop?
metarubricyte
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What does an eccentric nucleus mean?
off to the side of the cell
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Describe what a myeloblast looks like.
- large cell
- lots of pale blue cytoplasm
- round - oval nucleus
- 1 - 2 nucleoli
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Describe a progranulocyte.
- larger than a myeloblast, but can be smaller at times
- no nucleoli
- perinuclear clear zone
- small pinkish cytoplasmic granules
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What are small pinkish cytoplasmic granules called?
azurophilic granules
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Describe a myelocyte.
- smaller cell
- larger distince cytoplasmic granules
- nucleus is round to oval
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Describe a metamyelocyte
- indented, kidney shaped nucleus
- may see granules in cytoplasm
- mature into bands
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Describe a band.
nucleus has parallel sides
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What is the series of cells in monocytopoiesis?
- monoblast
- promonocyte
- monocyte
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Do we cell a lot of monocytes in bone marrow?
no
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What is the series of cells in lymphopoiesis?
- lymphoblast
- prolymphocyte
- lymphocyte
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What does a lymphoblast look like?
large lymph with nucleolus
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What does a prolymphocyte look like?
smaller than lymphoblast with no nucleolus
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If we see large numbers of lymphocytes in bone marrow, what could it indicate?
lymphoma or lympho-prolierative leukemia
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What can we confuse lymphocytes with?
rubricytes
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What are "free nucleus"?
skipocytes in bone marrow, can not count these as cell
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What breaks of megakaryocytes?
thrombocytes
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Describe a megakaryoblast.
- large cell
- multinuclear mass
- small amount of cytoplasm
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Describe a promegakaryocyte.
- large cell
- multinuclear mass
- more cytoplasm
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Describe a megakaryocyte.
- largest cell of the series
- multinuclear mass
- large amount of cytoplasm
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What is the difference in appearance between a osteoclast and a megakaryocyte?
- megakaryocyte is multinuclear but they are clumped together
- osteoclast has different distincy nuclei
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Which is seen more frequently in bone marrow, osteoclast or osteoblast?
osteoblast
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What does an osteoblast look like?
- oval, eccentric nucleus
- nucleoli
- large amount of basophilic cytoplasm
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What does a plasma cell look like?
- round cell
- round eccentric nucleus with clumpy chromatin
- perinuclear pale zone
- large amounts of basophilic cytoplasm full of vacuoles and russell bodies
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What turns into plasma cells?
B-lymphocytes
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What are russell bodies and where do we see them?
- antibodies
- in the cytoplasm of plasma cells
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What should always be done the same day as a bone marrow aspirate?
CBC
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How do we do a M:E ratio?
- add up # of white cells
- add up # of red cells
- divide and make a ratio
example: 275 white cells and 225 red cells...ratio 275/225 = 1.2:1
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What is the normal limit for a M:E ratio?
1 - 3:1
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What is considered an elevated M:E ratio?
ratio greater than 3
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What is an elevated M:E ratio due to?
increase in WBCs or decrease in RBCs
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What are the two situations that can cause an elevated M:E ratio?
- myeloid hyperlasia
- erythroid hypoplasia
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What do we expect to see on the CBC with a myeloid hyperplasia?
leukocytosis
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What do we expect to see on the CBC with a erythroid hypoplasia?
anemia
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What is considered a decreased M:E ratio?
ratio below 0.5 (example: 1:2)
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What is a decreased M:E ratio due to?
decrease in WBCs or increase in RBCs
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What are the two situations that can cause a decrease in M:E ratio?
- myeloid hypoplasia
- erythroid hyperplasia
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What do we expect to see on a CBC with myeloid hypoplasia?
leukopenia/neutropenia
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What do we expect to see on a CBC with a erythroid hyperplasia?
responsive anemia
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What other findings might we see on a bone marrow aspirate?
- no RBCs or WBCs
- increase in abnormal cells
- undifferentiated large cells
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What is it called when there are no RBCs or WBCs in the bone marrow?
aplasia
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What is aplasia?
lack of cell production
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What can aplasia be due to?
- immunosuppressive drugs (radiation, chemo)
- viruses (parvo, ehrlichia)
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What will need to be done if there we find aplasia in the bone marrow?
bone marrow transplant
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What are cells can become abnormal and increased in the bone marrow and what does it usually indicate?
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If we see undifferentiated large cells in the bone marrow (we can not tell them apart), what does that usually indicate?
some type of neoplasia
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What are myeloproliferative disorders?
- cell production out of control (preneoplasia)
- immature cells > mature cells
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What is multiple myeloma?
neoplasia of plasma cells
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What will show up on radiographs on an animal with multiple myeloma?
dark holes in the bone
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What is the main clinical sign of multiple myeloma?
lame
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What will we see in the bone marrow in an animal with multiple myeloma
plasma cells in the bone marrow
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Describe a mast cell.
purple cytoplasmic granules that cover up the nucleus
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Which ones are bands
the cells with the "u" shaped nucleus
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What is wrong?
erythroid hyperplasia
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What is wrong?
granulocyte hyperplasia
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What is this a group of?
osteoblasts (very eccentric nucleus)
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Identify the rubriblast.
the large round cell on the left
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What are these kinds of cells?
leukemic cells
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What are these cells? What is the cell in the middle?
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What is this indicate?
lymphoma
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What is this?
mast cell tumor
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What kind of cell is this?
megakaryoblast: multinuclear, small amount of cytoplasm
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What kind of cell is this?
megakaryocyte: large amounts of cytoplasm, multinuclear
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Which one is a metamyelocyte?
- the one on the right
- indented-kidney shaped nucleus
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What kind of cell is this?
Mott cell: full of vacuoles, russell bodies
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What is this?
multiple myeloma
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Identify the myeloblast.
- large cell on the left
- lots of pale blue cytoplasm
- round - oval nucleus
- 1 - 2 nucleoli
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What are these?
myelocyte: large distincet cytoplasmic granules, nucleus is round to oval
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Identify the myeloids and the erythroids.
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What is this?
osteoblast
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What is this?
osteoclast
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What is this?
osteoclast
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What is the cell in the middle?
plasma cell: round cell, round nucleus, eccentric nucleus
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What is this?
progranulocyte: no nucleoli, perinuclear clear zone, small pinkish cytoplasmic granules
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What is this?
promegakaryocyte: multinuclear mass, more cytoplasm
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What is this?
prorubricyte: no nucleolus, perinuclear clear zone, lacey chromatin
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Which ones are RBCs and which ones are WBCs
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What is at the pointer?
rubriblast: narrow rim of bright blue cytoplasm, round nucleus, 1 - 2 nucleoli
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What is wrong?
small cell lymphoma
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What is happening in this picture?
thrombocyte release
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What are these?
thrombocytes
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