Hematology

EDTA inversions

8

Heparin inversions

8

3.2% Socium citrate inversions

3-4

Formulations of EDTA

K2 EDTA

Concentraions for EDTA

1.5 mg/mL of blood

Concentrations for Heparin

15-20 units/mL

Formulations of Heparin

Ammonium, Lithium, Sodium Citrate

Formulations for 3.2% Sodium citrate

3.8% sodium citrate, westergreen ESR

Concentrations for 3.2% sodium citrate

A:C - 1:9

Action of Heparin

Binds Thrombin

causes swelling of platelets

EDTA

Cellular Clumping = Increase WBC, Decrease Plt

Heparin

Activation of platelets

3.2% sodium citrate

Mean platelet Volume (MPV)

Increase in 20% on 1st hour EDTA

Uses od EDTA

Routine hema

Uses of Heparin

Flow cytometry, plasma chemistry determination, osmotic fragility test (OFT), Blood gas studies

Uses of 3.2% sodium citrate

coagulation studies

Blood smear must be made

within 3 hrs. collection

EDTA in RT

6 hours

EDTA in 4 deg C

24 hours

Heparin NOT TO BE USED IN

Blood smear preparations

Coagulation studies

Prevents platelet AGGREGATION

EDTA

Insuffiicient EDTA

Clots

Excessive EDTA

👎🏻HCT, ESR

👍🏼MCHC, PLT

Lithium Heparin

Least interference in chemistry testing

Bluish smear bg with Romanowsky stain due to pH

Heparin

What color of AC?

SPS (sodium polyanethol sulfonate)

ACD ( acid citrate dextrose)

Yellow

K2 EDTA Tan

Lead determination

K2 EDTA Royal blue

Toxicology

Nutrition chemistry

Trace element determination

K2 EDTA with Gel (white)

Molecular diagnostic test

Pink K2 EDTA

Blood banking

Whole blood hema determination

Black 3.8% sodium citrate A:C

4:1

Black 3.8% sodium citrate uses

westergren ESR

3.2% sodium citrate A:C

1:9

3.2% sodium citrate uses

Coagulation Test

Light blue Citrate, Theophylline, Adenosine, Dipyridamole (CTAD) uses

Coagulation test

PF-4 and BTG or platelet fuction assay

Platelet Satellitosis/ Satellitism/Rosette in EDTA

cause pseudothrombocytopenia and pseudoleukocytosis

Two Glass Slide Method/ Manual Wedge Technique

30-45 degree angle

Coverslip Technique using Beacom’s Method

uses Glass-slide coverslip method

Coverslip Technique using Ehrlich’s Method

Two-coverslip metthod

Automated Machines

Cella vision hemaprep

Centrifugal (spinner type)

Coulter LH

Sysmex SP-10

Coulter LH

Slide maker & stainer

Sysmex SP-10

Slide maker & stainer

Fixative for EDTA

Methanol

Stain for EDTA

Wright/Wright-Giemsa Stain

Buffer pH 6.4

0.05 M Sodium Phosphate

Aged distilled water pH

6.4-6.8

Manual staining with Wright stain for how many minutes

1-3 minutes

Automated staining stain batch how many minutes

takes 5-10 mins

Automated staining examples

Midas

Hema-tek

Coulter LH

Sysmex Sp-10

Quick staining how many minutes

I min.

Staining of quick staining

modified wright or wright-giemsa stain filtered into a Caplin jar or staining dish

Quick staining buffer

aged distilled water

Ideal blood smear (wedge method)

2/3 or ¾ lenght of film slide finger shaped

THICK TO THIN AREA

Well-stained blood smear macroscopic color?

pink to purple

Well-stained blood smear microscopic RBcs color?

orange to salmon pink

Well-stained blood smear WBC nuclei color?

purple to blue

Well-stained blood smear neutrophil cytoplasm color?

pink to tan

Well-stained blood smear eosinophil granules color?

Bright-orange

POLYPHYLETIC THEORY

each blood cell limeage derived from ITS OWN UNIQUE STEM CELL

MONOPHYLETIC THEORY (WIDELY ACCEPTED)

all blood cells are derived from a SINGLE PROGENITOR STEM CELL TO PLURIPOTENTIAL STEM CELL

Mesoblastic/ Megaloblastic stage chief site?

Yolk sac

Hepatic chief site?

Fetal liver with: spleen thymus, lymph nodes

Medullary/ Intramedullary/ Myeloid chief site?

BM

What stage occurs the Primitive erythroblast?

In mesoblastic/ megaloblastic

Formed during 1st 2-8 weeks of life?

Mesoblastic/ Megaloblastic

Produce Hb in Mesoblastic or Megaloblastic

Gower 1

Gower 2

Portland

Predominant during this phase

Fetal Hb ( Hb F)

1st fully develop organs in fetus

Thymus

Major site of T cell production

Thymus

Produce B cells

Kidney & spleen

HEMATOPOIESIS STARTS

before the 5th month of fetal development

chief site of hematopoiesis

end of 24th week of gestation

Megakaryocyte the largest cell in normal BM

30-160 um

Macrophages size

40-50 um

mast cell size

12-25 um

WATERBUG/ COMET appearsnce, synthsize new bone matrix

Osteoblast

Large, multinucleated with size of > 100 um

Osteoclast

Total RBC mass circulating in the precursor in the bone marrow, RBC precursors

Erythron

RBCs in the circulation

RBC mass

DYNAMICS OF RBC creation and destruction

Erythrokinetics

Defective RBC precursor

Ineffective erythropoiesis

Decrease in RBC precursors

Insufficient Eryhtopoiesis

Macrocytic, Normochromic

Vit. B12 deficiency

Folate deficiency

Microcytic, Hypochromic

Sideroblastic anemia

Thalassemia

Iron deficiency anemia

Normocytic, Normochromic

Acute leukemia

Renal disease

BLUENESS of a cell part d/t acidic components that attract basic stain

Basophilia

Correlates with QUANTITY OF RIBOSOMAL RNA

Basophilia