Light Scatter	Gate 1-CD45/14	Gate 2-CD45/14

Gate 1-CD3/DR	Gate 2-CD3/DR

Gate 1-CD10/TdT	Gate 2-CD10/TdT

Gate 1-CD56	Gate 2- CD56

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Explanation and interpretation of Flow cytometry results
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Histology on Neck Mass
Special stains on neck mass

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• The neck mass demonstrates a normal sized lymphocyte population (gate 1) which makes up the majority of cells in this specimen. There is also a highly variable cell population that ranges from small to large in size and is more granular (complex) than the lymphocyte population (gate 2).  This variable population is a very minor population and has some similarities to the light scatter patterns seen in samples which are starting to lose structural integrity. This population is certainly not the first population that the technologist would examine (if they did at all) (see light scatter).

 
• The CD45/14 contour plot (gate1) confirms that the majority (87%) of these cells are of leukocyte origin.   The majority (78%) of cells in gate 2 are not leukocyte in origin or at least do not express CD45.  At this point the cells could still be debris (from degrading leukocytes) or stem cells, plasma cells, or other cells of non leukocyte origin.  The light scatter is not that expected for stem cells or plasma cells which would be a much less diverse pattern.  This leaves debris or non leukocyte as the most likely cell.  There are no monocytes/promonocytes present (see CD45/14).

 
• The DR/CD3 contour plot of gate one confirms the lymphocyte nature of these cells (66% CD3 - T cell receptor) and gate two again confirms the lack of T cells as well as B cells and monocytes (B and monocytes normally express HLA-DR). (see CD3/DR.

 
• The CD10/TdT plot confirms the lack of early precursor B and T cells as well as stem cells in this specimen (both gates are negative). (see CD10/TdT).

 
• Finally, the last contour plot is that of CD56 (homotrophic adhesion molecule) which is found on NK cells, some T cells, and neuronal cells. This antibody was added due to the fact that the pathologist found a few rosette clusters suggesting neuroblastoma.  In this case the large cell gate #2 was negative for CD45 (NK and T cells are positive for CD45) but positive for CD56. This suggests that these cells are, in fact, neuronal in origin. (see CD56).

 
• All of these results when taken together confirms the initial histological suspicion of neuroblastoma on the H&E slide. This case was usual in that the rosettes present were not abundant making the diagnosis difficult.  However, Flow cytometry and the special stains confirmed the neuronal nature of this specimen. 

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Histology on Neck Mass
Special stains on neck mass
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