Hematology: Platelet Clumping Issues
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Questions and Answers

What is the primary reason for performing a blood count using a 3.2% sodium citrate tube instead of using EDTA?

  • To ensure accurate measurement of platelet and leukocyte counts (correct)
  • To increase the platelet count significantly
  • To decrease the volume of blood needed for the test
  • To prevent artifacts during slide preparation

Which of the following describes a characteristic of pseudothrombocytopenia?

  • Low platelet counts that are misreported as elevated white blood cell counts (correct)
  • Elevated platelet counts resulting from sample contamination
  • High platelet count falsely reported due to inferior laboratory techniques
  • Inaccurate leukocyte counts due to overlapping platelet clumps

What is the effect of an incorrect angle when using the two-glass slide method for blood film preparation?

  • It has no effect on smear quality.
  • Both high and low angles result in unusable smears.
  • A high angle results in a thin smear, while a low angle results in a thick smear.
  • A low angle produces a thin smear, while a high angle results in a thick smear. (correct)

What is a disadvantage of using anticoagulant-free blood from a finger prick?

<p>The limited amount of blood makes it challenging to prepare multiple films. (C)</p> Signup and view all the answers

What is the ideal length of a blood smear film when using the wedge method?

<p>2/3 to 3/4 the length of the film (B)</p> Signup and view all the answers

What does an increase in hematocrit require when preparing a blood smear?

<p>Decrease the angle as low as 25° (C)</p> Signup and view all the answers

What is a common issue found with blood smears that have uneven distribution of white blood cells?

<p>Concentration of monocytes in the feathered edge (D)</p> Signup and view all the answers

Which of the following best describes the ideal shape of a blood smear?

<p>Finger-shaped with clear edges (C)</p> Signup and view all the answers

What is the effect of spreading the blood smear too slowly?

<p>Results in a thinner smear with poor distribution of WBCs (D)</p> Signup and view all the answers

What is one advantage of using the centrifugal (spinner) type preparation method?

<p>Creates a consistent preparation of blood cells. (A)</p> Signup and view all the answers

Which solution is NOT necessary for blood film staining?

<p>Distilled alcohol (C)</p> Signup and view all the answers

In Romanowsky-type stains, which component is primarily responsible for coloring the nucleus blue/purple?

<p>Methylene blue (D)</p> Signup and view all the answers

What is the main purpose of using fixative in blood film preparation?

<p>To preserve cellular morphology. (C)</p> Signup and view all the answers

Which automated instrument is primarily used for both slide making and staining?

<p>Coulter LH (D)</p> Signup and view all the answers

Why is it important to use non-anticoagulated blood for certain automated methods?

<p>It allows for a more even distribution of blood cells. (D)</p> Signup and view all the answers

When manually preparing a blood smear, what is the ideal time to leave the Wright's stain on the slide?

<p>1-3 minutes (D)</p> Signup and view all the answers

Which of the following statements is true regarding the Sysmex SP-10?

<p>It is used for both slide preparation and staining. (C)</p> Signup and view all the answers

What is a likely cause of eosinophil granules appearing gray in a blood smear?

<p>Buffer was too basic (D)</p> Signup and view all the answers

Which unusual finding in a blood smear could indicate a patient with increased lipid levels?

<p>Blue specks at the feather edge (B)</p> Signup and view all the answers

What appearance would indicate that the red blood cells in a blood smear might be improperly stained?

<p>Pale or red in color (C)</p> Signup and view all the answers

When examining a blood smear, what would a grainy appearance typically indicate?

<p>Improper slide preparation (B)</p> Signup and view all the answers

Which of the following is NOT a common problem encountered in blood smear staining?

<p>High fluorescence (D)</p> Signup and view all the answers

What should be avoided when using the 40x high-dry objective for WBC count estimation?

<p>Viewing the thick portions of the smear (A)</p> Signup and view all the answers

Which of the following factors influences the rejection of a blood smear film during microscopic examination?

<p>Presence of more than four times the usual number of WBCs at the lateral edges (A)</p> Signup and view all the answers

What is the primary use of the 100x oil immersion objective during a blood smear examination?

<p>To analyze nuclear details of leukocytes (D)</p> Signup and view all the answers

When estimating the WBC count using the 40x high-dry objective, how is the final count calculated?

<p>Average no. of WBCs per field multiplied by 2,000 (B)</p> Signup and view all the answers

What indicates a potential problem when observing RBCs in the feathered edge while using the 40x objective?

<p>RBCs appear macrocytic and flattened (D)</p> Signup and view all the answers

What is the estimated platelet count per µL if the average observed platelets in ten fields using the 100x objective is 150?

<p>3,000,000/µL (C)</p> Signup and view all the answers

During microscopy, which condition would require a separate process for transferring observations from the 10x objective to the 100x objective?

<p>Incompatibility of findings across different magnifications (D)</p> Signup and view all the answers

What characteristic should be noticed when examining the ideal area of a blood smear under the 10x low power objective?

<p>RBCs near each other but not overlapping (D)</p> Signup and view all the answers

Flashcards

EDTA-induced platelet clumping

EDTA, an anticoagulant, causes platelets to clump together, making them appear fewer than they actually are.

Pseudothrombocytopenia

A condition where a low platelet count is falsely reported due to platelet clumping by EDTA, not a true decrease.

3.2% Sodium Citrate

An anticoagulant that does not cause platelet clumping, used when platelet clumps are suspected.

Platelet clumps

Aggregation of platelets, often seen with EDTA, making platelet count measurement inaccurate.

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Anticoagulant-free blood

Blood collected without adding an anticoagulant, used for tests that require observing the blood's state.

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Two-glass slide method

A method for creating a blood smear by carefully placing one slide on top of another, spreading the blood sample thinly onto one slide.

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Pusher slide (spreader)

A slide used to spread the blood sample thinly across another slide in the two-glass slide method for creating a blood smear.

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Film slide

The slide onto which a thin blood smear is prepared using the glass slide method.

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30°–45° angle

The precise angle between the pusher and film slide. Too high or low leads to bad smears.

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Thick smear

A blood film that is too thick to analyze properly.

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Thin smear

A blood film that is too thin to analyze properly. Not enough blood.

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Blood smear size

A blood smear should be 2-3mm in size, 1cm away from label.

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Optimal spreader speed

The spreader speed must be neither too fast nor too slow. Proper technique is learned through practice.

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Hematocrit effect on smear angle

Adjust the angle (25°) to account for patient's hematocrit (PCV). Higher hematocrit means lower angle for thinner smears.

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Large smear

A blood smear that is thicker and longer than ideal.

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Small smear

A blood smear that is thinner and shorter than ideal.

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Fast smear speed

A blood smear made using a fast spreader speed results in thick smears.

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Slow smear speed

A blood smear made using too slow a spreader speed will result in thin smears and poor WBC distribution

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Longitudinal scanning

Scanning the blood smear along its length.

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Battlement (Rodak) scanning

Scanning the blood smear in a back-and-forth serpentine pattern.

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Ideal blood smear (wedge method)

A blood smear showing a gradual transition from thick to thin, taking up 2/3 to 3/4 of the slide; finger-shaped, with visible lateral edges and no irregularities or holes.

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Uneven WBC distribution

An issue where white blood cells are not evenly distributed throughout the blood smear.

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Feather edge issues

Areas with more segmented neutrophils, monocytes, and eosinophils at the edges of the blood smear.

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Center of film issues

Areas with more small lymphocytes at the center of the blood smear.

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CellaVision HemaPrep

A semi-automated device for preparing blood films, requiring no power source.

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Centrifugal blood smear

Blood film prep using a spinner, ensuring even distribution of cells, consistent preparation, and fewer smudge cells.

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Coulter LH

Automated instrument for slide making and staining process.

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Sysmex SP-10

Automated blood film stainers and analyzers.

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Blood film staining purpose

Evaluates the cellular morphology of blood cells for laboratory analysis.

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Methanol (fixative)

Preservative for blood films, toxic and must be handled carefully.

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Wright stain

Common blood film stain, combining methylene blue and eosin.

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Romanowsky-type stain

Blood stain containing Methylene Blue and eosin, commonly used to view blood morphology.

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Methylene blue

Basic stain that colors cell nuclei and other structures blue/purple.

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Eosin

Acidic stain that colors cytoplasm orange-red.

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RBC color (stained)

Orange to salmon-pink when properly stained.

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WBC nuclei color

Purple to blue when properly stained.

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Neutrophil cytoplasm color

Pink to tan when properly stained.

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Eosinophil granules color

Bright orange when properly stained.

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Blue/purple RBC

A sign of issues with staining, potentially due to an over-basic stain or incorrect buffer.

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Gray RBCs

Indicative of staining issues, potentially due to an inappropriate stain/buffer.

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Dark WBCs

A sign of staining issues, potentially due to an unsuitable staining solution.

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Gray Eosinophil granules

Poor staining, caused by problems in the staining process.

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Too pale/red RBCs

Indication of acidic buffer, potentially an over-acidic staining solution or buffer.

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Barely visible WBCs

A possible inadequate staining solution or rinsing.

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Bluer-than-normal blood film

Unusual finding, possibly elevated blood proteins (like in plasma cell myeloma).

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Grainy blood film

Unusual finding, possibly RBC agglutination.

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Holes in film

Unusual finding, possibly due to problems during preparation.

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Blue specks on film edges

Unusual finding, possibly elevated lipids or other elevated cell counts.

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10x Objective

Used for initial blood smear assessment (overall quality, color, cell distribution).

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Snowplow Effect

Excessive white blood cells concentrated at the edges of a blood smear.

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40x High-Dry Objective

Used for estimating total WBC count by counting leukocytes in 10 fields.

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WBC Count Calculation

Average WBC count per field x 2,000 = estimated WBC/µL (for 40x)

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50x Oil Immersion Objective

Higher magnification for detailed cell examination; different formula for WBC count estimation.

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Feathered Edge

Peripheral region of the blood smear where cells might be distorted or abnormal.

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Thick Part of Smear

Areas that are crowded with cells; avoid these regions on the blood smear.

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Rouleaux

Stacking of red blood cells; may affect WBC count reading.

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100x Oil Immersion Objective

Used for examining cell details (nucleus) and WBC differential, platelet count.

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Platelet Count (OIO)

Estimate platelet count by counting platelets in 10 oil immersion fields x 20,000.

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RBCs Per Oil Immersion Field

Approximate range (200-250) used for determining appropriate areas on the blood smear for assessment, affects platelet counting formulae.

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Study Notes

EDTA-Induced Platelet Clumping

  • EDTA (ethylenediaminetetraacetic acid) can cause platelets to clump together, appearing as large clumps, similar in size to white blood cells (WBCs).
  • This can cause a falsely low platelet count.

Pseudothrombocytopenia

  • A falsely low platelet count due to the effects of EDTA.
  • The incorrect counts occur because the anticoagulant EDTA causes clumping of platelets, making them appear fewer in numbers.

Pseudoleukocytosis

  • A falsely elevated white blood cell (WBC) count.
  • Large platelets might be misidentified as WBCs, leading to a higher WBC count as determined by a manual count.

Case Study

  • Initial platelet count was 90,000/mm³.
  • Initial leukocyte count was 15,000/mm³.
  • Platelet clumps were observed on blood smear.
  • Blood recollected using sodium citrate.
  • Second platelet count was 115,000/mm³.
  • Second WBC count was 11,000/mm³.
  • Correct final platelet count: 126,500/mm³.
  • Correct final WBC count: 12,100/mm³.

Anticoagulant-Free Blood (Finger Prick)

  • Advantages:
    • Performed at the patient's side, minimizing delays and complications.
    • Can sometimes prevent artifacts.
  • Disadvantages:
    • Platelets can clump, leading to inaccuracies.
    • Few well-prepared blood films can be made, when finger pricks have healed.

Blood Film Preparation Methods

  • Two-glass slide method (Manual Wedge Technique): Most frequently used
    • Materials: Pusher slide (aka: spreader), film slide
    • Procedure: The angle between the two slides should be between 30° and 45°, to create thiner or thicker smears.
    • Quality Control: The angle of the spreader determines smear thickness(too high leads to thicker smears, too low leads to thinner smear).

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Description

Explore the phenomena of EDTA-induced platelet clumping and its implications for blood count accuracy. This quiz covers pseudothrombocytopenia and pseudoleukocytosis through a detailed case study, illustrating the importance of proper anticoagulant use. Test your knowledge on interpreting laboratory results affected by these issues.

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