Hematology Laboratory testing- special stains

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In the Kleihauer-Betke test, a maternal blood smear is treated with acid and then stained with counterstain. The fetal cells contain fetal hemoglobin, which is resistant to acid and will remain pink. Since the calculated volume of fetomaternal hemorrhage is an estimate, how many additional RhIg vials need to be added for the dose?

1

The attached picture is from a blood smear stained with brilliant cresyl blue, a supravital stain. Select which cell(s) would be identified as reticulocyte.

Cell A

What is the Kleihauer-Betke test used for?

Differentiate between maternal and fetal red cells

Sudan III will confirm the presence of:

Fat

What is the principle of the Kleihauer-Betke test?

Fetal cells present in the mother's blood smear contain fetal hemoglobin (Hgb F). When treated with acid the fetal cells are resistant and will stain pink with the counter stain.

A 41-year-old male arrived at the clinic with complaints of fatigue, malaise, and overall discomfort. Upon testing, the patient's CBC results were notably abnormal with 30% blasts present along with anemia and thrombocytopenia. Bone marrow and cytochemical staining were subsequently performed. The cytochemical stains noted negative reactions to Myeloperoxidase and Sudan Black B but a positive reaction to alpha-naphthyl acetate. Which of the following conditions is most likely seen?

Acute Monocytic Leukemia

The following stain reactions were resulted on a suspected case of leukemia: Myeloperoxidase + Sudan Black B + Chloroacetate Esterase + Nonspecific Esterases - Which of the following is the correct interpretation?

Acute Myelocytic Leukemia

Cells that stain positive with acid phosphatase, and are NOT inhibited with tartrate, are characteristically seen in:

Hairy cell leukemia

This image depicts a brilliant cresyl blue stained blood smear. What inclusion bodies are shown in the erythrocyte indicated by the arrow (B)?

Hb H inclusions

A medical laboratory scientist working in Hematology was performing Myeloperoxidase staining on a suspected AML case. They found positive reactions of the segmented neutrophils. What would be the next step?

Inaccurate result, only blast cells are to be examined for reaction

Cells, as shown in this iron-stained bone marrow preparation, are found in each of the following conditions EXCEPT:

Iron deficiency anemia

Which laboratory procedure may be used to assist in differentiating chronic myelocytic leukemia (CML) from a leukemoid reaction?

LAP stain

If a patient has a white blood count of 40,000/µL, what would be the MOST useful staining procedure to distinguish between bacterial infection and chronic myelogenous leukemia (CML)?

Leukocyte alkaline phosphatase (LAP)

Which of the following stains is used to differentiate a neutrophilic leukemoid reaction from chronic myelogenous leukemia (CML)?

Leukocyte alkaline phosphatase (LAP)

If a patient's WBC is 50,000/mm3, what test should be ordered to determine if this is a leukemoid reaction or a chronic myelocytic leukemia?

Leukocyte alkaline phosphatase stain

Laboratory results obtained on a 47-year-old patient were as follows: WBC 51.2 x 103/µL RBC 4.27 x 106/µL HGB 12.6 g/dL HCT 36.9% PLT 403 x 103/µL Differential: 50% segmented neutrophils 20% bands 12% lymphocytes 7% monocytes 1% eosinophils 9% metamyelocytes 1% myelocytes RBC morphology: normocytic, normochromic WBC morphology: moderate toxic granulation, slight Dohle body Which test would be MOST helpful in establishing a diagnosis in this case?

Leukocyte alkaline phosphatase stain (LAP stain)

Which of the following statements is true regarding the staining protocol used for bone marrow smears compared to that for peripheral blood smear?

Longer staining time should be used to allow thorough staining

The peroxidase stain is useful in differentiating between which cells?

Lymphocytic and granulocytic cells

Esterase stains are used to help differentiate which of the following?

Monoblasts from myeloblasts

In which stain will auer rods have the most intense reaction?

Myeloperoxidase

By utilizing a Prussian Blue stain, which of the following red blood cell inclusions would be identifiable if present? Note: Upper image = Wright-Giemsa stain Lower image = Prussian Blue stai

Pappenheimer (siderotic) bodies

What are the red cell inclusions in this split frame

Pappenheimer bodies

When a few small, purple inclusions are found in erythrocytes, they can be confirmed as containing iron by:

Performing a Prussian blue stain

In addition to a Romanowsky stain, which one of the following would be useful in the routine evaluation of a bone marrow?

Prussian blue

Which of these methods could be used to differentiate between Howell-Jolly bodies and Pappenheimer bodies and what results would be noted?

Prussian blue stain: Pappenheimer bodies stain blue but Howell-Jolly bodies do not stain blue

For which determination is the brilliant cresyl blue stain used MOST often?

Reticulocytes

What is the cell indicated by the arrow in this Prussian blue stain?

Ringed sideroblas

Which of the following terms most accurately identifies the cell indicated by the arrow in this bone marrow slide that was stained with Prussian blue iron stain?

Ringed sideroblast

The image is of a peripheral blood smear stained with Prussion blue stain. What are the inclusions seen in the RBC indicated by the arrow?

Siderotic granules

Methylene blue stain works by

Staining cellular components that have an acidic pH such as ribosomes, DNA, and RNA

What is the principle of the New Methylene Blue test?

Staining cytoplasmic RNA, mitochondria, and ribosomes

What is the principle of the Kleihauer-Betke test?

The Kleihauer-Betke test capitalizes on the resistance of fetal hemoglobin to acid treatment.

Which of the following is considered to be the stain of choice for evaluation of a peripheral blood smear to include red cell morphology as well as a white cell differential count?

Wright

What stain is most commonly used in the hematology laboratory?

Wright stain

Pappenheimer bodies (or siderotic granules) in erythrocytes can be detected using the following stain/s.

Wright's stain and Prussian blue stain


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