White Blood Cells

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Presentation Transcript

White Blood Cells:

White Blood Cells Chapter 2

“For the life of all flesh is the blood thereof” Leviticus 17:14:

“For the life of all flesh is the blood thereof” Leviticus 17:14

Learning Objectives:

Learning Objectives Understand the importance of a WBC count and differential Describe the procedure for a WBC manual count Describe the procedure for a WBC differential count Identify types of WBC’s Understand the function of each type of WBC

The Complete Blood Count (CBC):

The Complete Blood Count (CBC) Total RBC count PCV Plasma Protein Concentration Blood film examination: differential WBC count , erythrocyte and leukocyte morphology , platelet estimation Reticulocyte count Hemoglobin concentration Erythrocyte indices

Manual WBC count:

Manual WBC count Unopette system (Becton, Dickinson) Pipette, reservoir Provides a specific dilution ratio Several different models with different dilution ratios NO LONGER MANUFACTURED

Manual WBC count:

Manual WBC count Leukocheck Replaces the Unopette Same procedure and dilution

Hemocytometer:

Hemocytometer Counting chamber Determine # of cells/ µl or mm 3 Neubauer rulings

Neubauer ruling (grid):

Neubauer ruling (grid) 2 identical sets of grids Each grid holds a precise amount of sample Can calculate the # of cells per µl of blood

Hemocytometer:

Hemocytometer

Manual WBC count - Procedure:

Manual WBC count - Procedure Procedure Blood added to diluent (lysing solution) Wait 10 minutes Invert to mix cells Convert to dropper Apply coverslip Charge chambers (fill wells) Place on microscope under 10x objective Count cells 9 squares or 4 corner squares dependent upon dilution (Unopette or Leukocheck)

WBC count video:

WBC count video

Double Ruling Method:

Double Ruling Method Counted if they are touching the top or the left line Not counted if they are touching the bottom or right line

Calculations:

Calculations Number of cells counted from both sides are added and divided by two Divided by dilution ratio and the volume factor that represents the volume of nine squares Example: 80 +76 78 average 78 divided by 1/100 dilution x 0.9 = 8.6 x 10 3 µl = 8,600 WBC/µl blood

Calculations:

Calculations OR Mathematic approximation of this number would be multiplying the average by 110 Example: 78 x 110 = 8580 WBC/ µl blood

Preparation of Blood Films:

Preparation of Blood Films Used to perform the differential WBC count ; estimate platelet numbers; and evaluate the morphologic features of WBCs , RBCs, and platelets

Preparation of Blood Films:

Preparation of Blood Films Wedge smears Drop of blood Back slide into drop and push forward at an angle Air dry

Blood Films:

Blood Films

Blood Films:

Blood Films

Blood Films:

Blood Films

Preparation of Blood Films:

Preparation of Blood Films Staining Blood Films Distinguish individual cells Identify abnormal cellular characteristics Romanowsky-type stains Wright’s stain Wright-Giemsa stain Diff-Quick, Stat Stain, CAMCO Quick Stain, Protocol Hema3

Stains:

Stains Fixative – 95% methanol Eosin Methylene blue Staining problems

Performing the Differential Cell Count:

Performing the Differential Cell Count Automated Cell counters? Consistent method Scan entire slide Identify feathered edge and then look for monolayer Focus on one field in the monolayer with oil immersion Count 100 WBCs, identify, and record as percentage.

Areas of slide:

Areas of slide

Definitions:

Definitions Relative WBC count - % of each type of WBC Absolute value – multiply total WBC count by % of each cell type Example – 80% neutrophils x 6000 WBC/ µl = 4800 neutrophils/µl blood

Definitions:

Definitions -penia – Decreased number of cells in the blood -philia or –cytosis – Increased number of cells in the blood Left shift – Increased number of immature neutrophils in the blood Leukemia – Neoplastic cells in the blood or bone marrow Leukemoid response – Can be mistaken for leukemia

Performing Count:

Performing Count Cell Counter WBC Bands Neutrophils Lymphocytes Monocytes Eosinophils Basophils

Leukocytes:

Leukocytes Granulocytes Neutrophils Eosinophils Basophils Agranulocytes Lymphocytes Monocytes

Neutrophils:

Neutrophils 1 st line of defense in infection Initiation of inflammatory response Phagocytosis

Neutrophils:

Neutrophils

Band Neutrophil:

Band Neutrophil

Hyposegmented:

Hyposegmented

Hypersegmentation:

Hypersegmentation

Dohle Body:

Dohle Body

Toxic Granulation:

Toxic Granulation

Bar Body:

Bar Body

Basophilic Cytoplasm:

Basophilic Cytoplasm

Ehrlichia Morula:

Ehrlichia Morula

Eosinophil:

Eosinophil Allergies Parasitic infections

Eosinophil:

Eosinophil

Vacuolated Eosinophils:

Vacuolated Eosinophils

Feline:

Feline

Equine:

Equine E

Domestic Ruminant:

Domestic Ruminant

Bird & Reptiles:

Bird & Reptiles E E

Basophil:

Basophil Similar to mast cells Acute inflammatory reaction Allergies and parasites

Basophils:

Basophils Feline Canine

Cat:

Cat

Equine:

Equine

Domestic Ruminants:

Domestic Ruminants

Lymphocytes:

Lymphocytes Immune response B-lymphocytes produce antibodies T-lymphocytes involved in cell-mediated immunity

Lymphocytes:

Lymphocytes

Reactive:

Reactive

Atypical:

Atypical

Azurophilic Granules:

Azurophilic Granules

Birds:

Birds

Monocytes:

Monocytes Macrophage when in the tissues Phagocytosis – handles those not handled by neutrophils May indicate chronic infections

Monocytes:

Monocytes

Monocytes:

Monocytes

Smudge Cells:

Smudge Cells

“The righteous man regardeth the life of his beast” Proverbs 12:10:

“ The righteous man regardeth the life of his beast” Proverbs 12:10