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BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013

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Page 1: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

BIOT 309: BLOOD SMEAR PRACTICAL

Feb., 2013

Page 2: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Microscopic Views

Bird Blood

Cat Blood

Dog Blood

Fish Blood

Frog Blood

Snake BloodHuman Blood

Horse Blood

Page 3: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Maslak, P. ASH Image Bank 2008;2008:8-00067. Copyright ©2008 American Society of Hematology.

ASH Teaching Slides: Blood Cells

Normal blood smear. The four larger cells shown are called granulocytes, a type of white blood cell.

Page 4: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Photomicrograph of a Blood Smear

Figure 10.2

Page 5: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Cells you will see

• Red blood cells, erythrocytes, no nucleus, transport O2

• Platelets, deliver fibrin• White blood cells, leukocytes: complete

cells with a nucleus and organelles, defenders against disease,

Page 6: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Cell Differentiation and Lineages

• Granulocytes–Granules in their

cytoplasm can be stained– Include neutrophils,

eosinophils, and basophils

Figure 10.4

Page 7: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Eosinophiles

• Function: Phagocytosis of antigen-antibody complexes; allergens

• Release enzymes to weaken or destroy parasites such as worms.

• Appearance: Nuceus has 2 large lobes

Page 8: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Basophil

• Appearance:

• Function: Secretes Histamine,( a vasodilator)• Secretes heparin ( an anticoagulant)

Page 9: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Neutrophils

Appearance• Nucleus usually with 3-5 lobes in S-C shaped

array

Functions: Phagocytosis of bacteriaRelease of antimicrobial chemicals

Page 10: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Precaution: You need the right size drop to get a good smear

• Too large drop = too thick smear too many cells• Too small drop = too thin smear too few cells

• Thick and thin also depend on (de)hydration, hematocrit (anemia)

Peripheral Smear Preparation

Page 11: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

2. The pusher slide is held securely with the dominant hand in a 30-45 deg angle.- quick, swift and smooth gliding motion to the other side of the slide creating a wedge smear

Peripheral Smear Preparation

Page 12: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Peripheral Smear Preparation

Page 13: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Precautions: Ensure that the whole drop of blood is picked

up and spread Too slow a slide push will accentuate poor

leukocyte distribution, larger cells are pushed at the end of the slide

Maintain an even gentle pressure on the slide – not doing so leads to ridges and valleys

Keep the same angle all the way to the end of the smear.

Peripheral Smear Preparation

Page 14: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Peripheral Smear Preparation

• Allow the blood smear to thoroughly dry – thicker will take longer

• Fix the blood smear with 70% methanol for 5 seconds; blot slide edge: let surface thoroughly dry

• Stain cells with Giemsa or Wright stain for 5’ - both are available; write in notebook which one you use – they color cells differently

• Dip slide in distilled water, 3 – 5 times• Blot edge; thoroughly dry

You are now ready to observe without and with microscope!!!

Page 15: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Feature of a Well Made Wedge Smear• Smear is 2/3 or ¾ the entire slide• Smear is finger shaped, very slightly rounded at

the feathery edge: widest area of examination• Lateral edges of the smear visible• Smear is smooth without irregularities, holes or

streaks• When held up in light: feathery edge should show

rainbow appearance• Entire whole drop of blood is picked up and

spread

Page 16: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood
Page 17: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

• See shape, internal morphology, comparative size, count cells, identify disease conditions

Page 18: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Peripheral Smear Examination• Visual – no microscope: record in lab notebook– Accurately draw both smears– Measure length and width in mm– Assess overall quality, i.e., • Does it have a feathery edge? Fingerprint shape? • Quality of the staining –also assess at

• Microscopic:• 10x Objective record in lab notebook– Observe types and distribution of the cells – use systematic

approach, i.e., serpentine• where do you find red blood cells; are they easily distinguished,

i.e., separated• Check lateral edges for WBC (white blood cells) distribution; do

you see them distinctly• Quick sketch of 2 different fields; label and describe cells

Page 19: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Choose the best areas for assessment

• Systematic search• Back and forth serpentine or battlement

track patterns are preferred – move the slide mount

Page 20: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Peripheral Smear Examination• Microscopic:• 40x Objective this week and next

1. Determine correct area to view – cells spread apart but not too much

Optimal Assessment Area:1. RBCs are uniformly and singly distributed2. Few RBC are touching or overlapping3. Normal biconcave appearance

Page 21: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

• Too thin

Peripheral Smear Examination

Too thick

Page 22: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Assessment at 40X

• Pick 5 fields that show different types of cells and sketch all cells; describe these cells in your own words

• Determine which cells you are seeing

Page 23: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Before you leave at 9:50

• Make sure your slides have your student ID#• Give to Younis; he will save for next week• Clean up your area

Page 24: BIOT 309: BLOOD SMEAR PRACTICAL Feb., 2013. Microscopic Views Bird Blood Cat Blood Dog Blood Fish Blood Frog Blood Snake Blood Human Blood Horse Blood

Final Assessment – Next Week

• Microscopic:• 100x objective, oil immersion