Suspect Flagging NE Blasts Imm NE 2 Imm NE 1 LY Blasts Variant LY MO Blasts NRBCs Depending on the Mean and SD of each parameter in the V, C and S, the flagging is programmed.
Suspect messageDescription Abnormal HemoglobinPattern characteristic of specimen with abnormal hemoglobin clearing observed during retic analysis Cellular InterPattern consistent with NRBC detection during a CBC only cycle. This Cellular Interference Suspect msg is not associated with a Review (R) flag Dimorphic RedsEvidence of the presence of at least two populations of red cells Giant PlateletsPatterns characteristic of specimen containing Giant Platelets Imm GransPattern characteristic of specimen containing: a) metamyelocytes and myelocytes and/or promyelocytes, or b) myelocytes and/or promyelocytes without metamyelocytes. Left ShiftPattern is characteristic of specimen containing metamyelocytes, but without myelocytes, promyelocytes, or blasts LY BlastBlasts in the Lymphocyte region of the dataplot MO BlastBlasts in the Monocyte region of the dataplot NE BlastBlasts in the Neutrophil region of the dataplot NRBCCBC & Diff pattern is characteristic of specimen with NRBCs.This Suspect msg applies when the NRBC analysis is disabled but a CD/CDR cycle is run. RBC Frag/MicrocytesThe specimen may contain red cell fragments &/ some microcytic red cells Red Cell AgglutRed cells may be clumped or display rouleaux on peripheral smear Sickled CellsPattern characteristic of specimen containing irreversibly sickled cells Variant LYPattern characteristic of specimen with variant lymphs, including mature lymph such as those observed viral infections, as well as immature&/ abn lymph
System MessageDescription Abn Diff PatternUndefined abnormal Diff pattern observed during Diff analysis. Cellular InterPoor separation between a WBC population and interference. WBC correction was performed as a best estimation, and the WBC was flagged with R MCV Inter: PLTInterference with MCV, RBC, and RDW and RDW-SD due to PLT. MCV Inter: WBCLow confidence in MCV due to interference from high WBC level MO-NE OverlapPopulation labeled as neutrophils appeared in the monocyte region during Diff analysis NE-EO OverlapNeutrophil and eosinophil populations were shifted or overlapped during Diff analysis Platelet ClumpsPattern is characteristic of specimen containing platelet clumps. RBC-PLT OverlapInterference with larger platelets; may occur with the Giant Platelet Suspect message. RET-RBC OverlapThe retics could not be clearly separated from the mature red cells Excessive DebrisThe number of debris events too high compared to white events during Diff analysis Excessive Low DC Events Too many low DC events during NRBC analysis Flow Cell Clog: DHardware detected a flow cell was clogged during Diff analysis
Haemopoiesis Production of RBCs, WBCs & Platelets
Bone Marrow Is the Factory that produces and Designs blood corpuscles from precursor cells This production is at a regular baseline level to maintain normal levels replacing the dead corpuscles that completes their life span – Red cells (90-120 days), Platelets (7-10 days), Neutrophils (few min to not more than one day) When the need arises the production is increased. The Design is to Fit the function. It utilizes structural elements of the marrow environment
Red blood cells Function Carries Haemoglobin - Tranport blood gases - Maintain blood pH Proper structure, size and shape - Flexibility to get to every tissue and cell. - Biconcave shape allows collapse
Neutrophil Function 1 st line of defence against infection Good Phagocytic cell ability to move well pseudopodia forming phagosomes Lots of cytoplasm to – accomodate phagocytosed – pack with cytotoxic lysosomal granules
Eosinophils Function Designed like Neutrophil but unable to have pseudopodia, phagocytosis or phagosomes due to large granules David vs Goliath Kills large organisms even more than 10 times its size – by spitting the granules instead of phagocytosing→ increasing toxic oxygen radicles – frustrated phagocytosis Anti-allergic – destroys histamin
Basophils/Mast cells Function Causes allergy, - contains large granules that contain Histamin and heparin
Monocytes/Macrophage/Reticulum cell Function Phagocytosis Lot of cytoplasm and all capabilities for killing organisms
Lymphocyte Function Antibodies and Cytokines production Active protein sysnthesis Therefore the cell has more nuclei and very little cytoplasm Not to store but to secrete
Platelets (Thrombocytes) Function Provides a lot of necessary procoagulant surface for clotting in Haemostasis Obtained from megakaryocyte which has the same surface - but 1,000 platelets from 1 megakaryocyte provides 10,000 times the surface than one megakaryocyte
Time line Maturation time from Proerythroblast to Reticulocyte takes 5 days Maturation from Myeloblast to Neutrophil takes 7 days At times of need the days are shortened. Shortens to 1 day in case of Neutrophils – Toxic changes – immature granules – Dohle bodies – retains the color of immature cytoplasm – Macro
ROC ANALYSIS AND CUT OFFS FOR NEUTROPHIL PARAMETERS FOR MDS NEUTROPHIL PARAMETERS sensitvityspecificityCut offAUCP value NEU MEAN VOL 35.997.4<137.90.5820.2 NEU MN VOL SD 69.284.2>22.50.7180.0008 NEU MEAN COND 48.797.4<147.70.5320.67 NEU MN COND SD 74.492.1>7.040.8860.0001 NEU MEAN SCATTER 61.494.7<1420.7190.0006 NEU MN SC SD 87.284.2>11.60.8930.0001
MDS on Cell counter The mean cell scatter (MNES) was significantly lower. Distribution of Cell volume (DNEV), Conductivity (DNEC) as well as Scatter (DNES) of neutrophils in the MDS group were significantly higher. Thus the cell counter identifies Neutrophil hypogranularity which could be obvious on smears depending on the stain used. In addition it identifies Neutrophil Anisocytosis (Variation in Cell size – DNEV), Neutrophil Anisogranularity (Variation in Granularity – DNES) and also Variation in cytoplasmic/nuclear structure among Neutrophils (DNEC). These features are not obvious on smears.
S-10;n-20;L-60;vL-10; 2.0 So very steep due to lesser small plts but peak continues after 10 at same amplitude Even after 10 - like there are two population Very Large population is significant – Creating a major hiatus 1016140199 Case -1
S 10;N10;L60;VL20; 2.5 – same scenario like previous case 1016140179 Case -4
Platelet on smear 0.5 – mostly small plts Peak before 10fl. Major Hiatus created by clumps and very large platelets - so curve not fitted – but has not significantly impacted platelet count since they are few 1016140503 Case -5
Platelets on smear 0.8 – same scenario as previous case 1016140406 Case -7
S0;N0;L80;VL20; 0.5 MCV70.7 VL + small RBCs Lesser platelets with more counts 1016140671 Case -11
20 year old pregnant female with fever for 1 month
WBC CBC histogram showed the presence of population below 35fl.
VCSVCS VCSVCS Malaria diagnosis ok I II III IV Normal RPD I
ParameterRulesComments PLT< 50,000 (irrespective of MPV) 50,000- 100,000 MPV >11.5 Slide Review NO Slide Review HGB<7g/dlSlide Review MCHC>35.8Slide Review MCV <70 and/or MCH <26 Slide Review MCV <70 and/or MCH <26 RDW > 22No Slide Review Lymph%>50 (adults) ; >60(child)Slide Review NRBCAny valueSlide Review Retic% & Retic #> 10% & > 0.1Slide Review All Suspect and System FlagsSlide review for all the following Newer Flags based on Ref Range on CPD Lymphocyte Microcytosis Neutrophil Macrocytosis Lymphocyte anisocytosis Current Validation Criteria (currently in use after consensus)