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Platelet Electron Microscopy: Utilizing LEAN Methodology to Optimize Laboratory Workflow
1Norton Children's Hospital, Louisville, Kentucky.
Background:
Platelet electron microscopy (PEM) is the gold standard methodology for diagnosing storage pool disorder, defined as a paucity of delta granules, alpha granules, or both. PEM literature is limited with few published resources and without well-developed interlaboratory standardization for the preparation and examination of platelet samples.
Methods:
Whole mount (WM) dense body (DB) counts for 300 pediatric cases were reviewed to determine whether counting fewer platelets could yield the same results. For 6 cases, DB average was determined on the day of WM preparation and on 2 consecutive days. Both WM and thin section (TS) preparations were examined for all cases.
Results:
Employing LEAN methodology, an algorithm was developed to markedly decrease the number of platelets counted and still ensure accurate results. WMs decay with time; a statistically significant difference in DB counts was noted between day 0 and day 1 (p < .1). Twelve of 300 cases required both WM and TS preparations for a complete diagnosis.
Conclusion:
It is possible to maintain accuracy and decrease 100 platelet DB counts by >75%. WMs must be counted on the day they are prepared to avoid false paucity of DB secondary to sample decay. An accurate evaluation of platelet morphology requires both the WM and TS techniques.
Insights
Platelet electron microscopy (PEM) can be optimized by reducing platelet counts. Whole mount preparations must be analyzed promptly to ensure accurate dense body (DB) counts for diagnosing storage pool disorders.
Area of Science:
- Hematology
- Microscopy
- Diagnostic Pathology
Background:
- Platelet electron microscopy (PEM) is the gold standard for diagnosing storage pool disorders.
- Limited literature and lack of standardization hinder PEM interlaboratory consistency.
Purpose of the Study:
- To optimize PEM by reducing the number of platelets analyzed.
- To establish standardized methods for accurate platelet morphology evaluation.
Main Methods:
- Reviewed 300 pediatric whole mount (WM) cases for dense body (DB) counts.
- Assessed DB counts on consecutive days for 6 cases.
- Examined both WM and thin section (TS) preparations for all cases.
Main Results:
- Developed a LEAN methodology algorithm to decrease platelet counts while maintaining accuracy.
- Observed statistically significant decay in WM DB counts over time (p < .1).
- 12 of 300 cases required both WM and TS for diagnosis.
Conclusions:
- Accuracy can be maintained while decreasing platelet DB counts by over 75%.
- Prompt analysis of WM is crucial to prevent false-negative results due to sample decay.
- Accurate platelet morphology assessment necessitates both WM and TS techniques.
