Preparation and Staining of Peripheral Blood Films

Question 1

Peripheral blood films can be made directly from

Answer 1

Skin puncture
Tube of EDTA-anticoagulated venous blood

Question 2

Best choice for PBS

Answer 2

Anticoagulant-free blood

Question 3

With a skin puncture, the phlebotomist must remember to

Answer 3

wipe away the first drop of blood

Question 4

Some patients’ blood undergoes an in vitro phenomenon called __________ when anticoagulated with EDTA.

Answer 4

platelet satellitosis

Question 5

Describe platelet satellitosis

Answer 5

The platelets surround or adhere to neutrophils

Question 6

When platelet adhere to neutrophils it potentially causes

Answer 6

Pseudothrombocytopenia

Question 7

Can result from EDTA induced platelet clumping

Answer 7

Low platelet counts
Falsely increased WBC count (pseudoleukocytosis)

Question 8

Manual Blood Film Preperation

Answer 8

Coverslip method
Wedge type

Question 9

grease free, square coverslip 22 sq.mm.
superior WBC distribution

Answer 9

Coverslip method

Question 10

Coverslip method should use

Answer 10

Use of two 1x3 in glass slides

Question 11

Demonstrate coverslip method

Answer 11

A drop of blood placed on the center of the lower slide,
then the top slide is placed on the blood drop so that the ends of the 2 slides overlap slighty. Then the 2 slides are pulled apart in a smooth, parallel motion.

Question 12

Variations of the ______ are the most widely used for smear preparation.

Answer 12

wedge technique

Question 13

Wedge type smear is termed as

Answer 13

Wedge, spreader-slide or push wedge

Question 14

Automated Blood Film Preparation

Answer 14

Wedge (Push) Type
Centrifugal (Spinenr) Method
Automated Slide Makers/Stainers

Question 15

Portable semi-automatic instrument that simulates the manual spreader-type technique

Answer 15

Wedge (Push) Type

Question 16

Automated instruments that utilize centrifugation to spread a monolayer of whole blood on a 1x3 inch glass slide

Answer 16

Centrifugal (Spinner) Method

Question 17

Smear preparation, stain, differential count report, platelet estimate, captures digital image, barcoding

Answer 17

Automated Slide Makers/Stainers

Question 18

The film is _____________________ the length of the slide

Answer 18

two thirds to three fourths

Question 19

The film is ___________, very slightly rounded at the
___________, not bullet shaped
o This provides the widest area for examination.

Answer 19

finger shaped
feather edge

Question 20

The ________ of the film are visible.

Answer 20

lateral edges

Question 21

The film is smooth with a gradual transition from

Answer 21

Thin to thick areas

Question 22

The film is smooth without irregularities,

Answer 22

no waves, no holes, or streaks and no bubbles.

Question 23

When the slide is held up to the light, the thin portion
(feather edge) of the film has a

Answer 23

“rainbow” appearance.

Question 24

It should allow at least ___ low power fields in which ____ of
the RBCs do not overlap.

Answer 24

10, 50%

Question 25

How do we know that it is not the area for differential count?

Answer 25

Rouleaux formation is the linking of RBCs into chains resembling stacks of coins

Question 26

best area for starting differential leukocyte count

Answer 26

Monolayer (middle part)

Question 27

What is differential leukocyte count?

Answer 27

100 WBCs to be differentiated

Question 28

UNACCEPTABLE PERIPHERAL BLOOD FILMS

Answer 28

Chipped, rough edge Hesitation in forward motion Drop of blood too small Dirt or grease on the slide (elevated lipids in blood specimen)

Question 29

The faster the smear

Answer 29

The shorter the smear

Question 30

More amount of blood in smear will produce

Answer 30

A thicker longer smear

Question 31

Widely used in diagnosis of blood parasites, particularly malaria (10x thickness of normal smears)

Answer 31

Thick Blood Smear

Question 32

Accepted thickness of thick blood smear

Answer 32

If you are able to read through the fine print of the newspaper

Question 33

Stains for thick blood smear

Answer 33

Giemsa Stain Field’s Stain

Question 34

Stain for both thin and thick smears to examine blood for malarial parasite

Answer 34

Giemsa Stain

Question 35

What is it that you should not forget about giemsa? What makes it different from Wright’s when all are actually Romanowksy stain?

Answer 35

prepared by diluting Giemsa stain powder in a buffered bacteria., Giemsa is used for biological specimen like bacteria and parasite. Wrights Stain is used for non-blood specimen

Question 36

Histological method for staining thick blood smears for malarial parasites.

Answer 36

Field’s Stain

Question 37

corresponds to a drop of blood spread over a circle approx.

Answer 37

2 cm in diameter

Question 38

The blood smears are mostly done for

Answer 38

Differential Leukocyte Count

Question 39

Essentially all specimens received for routine testing in the hematology section of the laboratory have been collected in _________topped tubes.

Answer 39

Lavender (purple) topped tubes.

Question 40

Lavender topped tubes contain

Answer 40

Ethylene-diaminetetraacetic acid (EDTA)

Question 41

EDTA anti-coagulates the blood by

Answer 41

Chelating calcium that is essential for coagulation

Question 42

_______________ is often preferred to the powdered form because it mixes more easily with blood.

Answer 42

Liquid tripotassium EDTA

Question 43

why do we need to preserve the morphology of the RBCs and the amount of the analytes at the time of collection

Answer 43

To prevent unwanted changes that occurs when blood is collected from the body. And in order for it to represent the exact conditions of the specimen at the time of collection

Question 44

Fixation and staining of smears is done using the ff.

Answer 44

3-5 mins absolute methyl or ethyl alcohol 1-2 mins formalin solution in 50% alcohol

Question 45

Principle of Staining

Answer 45

Acidic dyes unite with the basic components of the cell (cytoplasm) and conversely basic stains/dyes are attracted to and combine with the acidic parts of the nucleus (e.g. nucleic acids)

Question 46

First to use simple aniline dyes, first in sequence then pre- mixed acidic-basic stains (neutral dyes) = triacid: orange g, acid fuchsin, methyl green

Answer 46

Ehrlich

Question 47

The precipitate formed after mixing eosin and methylene blue has combined staining properties of the parent dyestuffs

Answer 47

Jenner (1889)

Question 48

He said that this precipitate that formed has improved staining capacity

Answer 48

Jenner (1889)

Question 49

Defined as a stain containing MB (methylene blue) and/or its products of oxidation and a halogenated fluorescein dye usually eosin;

Answer 49

Romanowsky (1890)

Question 50

The polychromed methylene blue has wider range of staining capability o It is able to stain

Answer 50

cell nuclei and platelet granules

Question 51

Most of the Romanowsky stains are prepared with _________ so that they act as a fixative as well as the cellular stain.

Answer 51

Methyl alcohol (Methanol)

Question 52

There are 4 different types of Romanowsky stains commonly used in Hematology laboratory for staining the blood cells:

Answer 52

o Wright’s Stain o Giemsa Stain o Leishman Stain o Field’s Stain

Question 53

Azure compounds with methylene blue and eosin, aqueous stain Best for inclusion bodies

Answer 53

Giemsa (1904)

Question 54

Giemsa In combination with Jenner or May Grunwald stains it constitutes

Answer 54

“Panoptic stain”

Question 55

An_________ means it has no alcohol

Answer 55

accustain

Question 56

Constitutes combination of a Romanowsky stain with another stain; such that it improves staining of cytoplasmic granules and other bodies: ▪ Jenner-Giemsa ▪ May Grunwald-Giemsa

Answer 56

Panoptic Staining

Question 57

is a mixture of Methylene blue, and Eosin dye, prepared in Alcohol medium and diluted with buffer or distilled water during staining procedure.

Answer 57

Leishman stain

Question 58

The Leishman stain is one of the best stains for routine blood stain to stain the Peripheral blood smear for the examinations of blood film under the microscope and is satisfactory for malaria and other blood parasites. o ___________ gives better results in parasitic studies.

Answer 58

Leishman stain, Giemsa stain

Question 59

It stains the different components of blood in a range of shades between red and blue. • It is based on a methanolic mixture of “polychromed” Methylene blue and eosin

Answer 59

Leishman stain