U2 LAB: PBS PREPARATION Flashcards by maki 🐈‍⬛

Q

Evaluation of peripheral blood film is an intergal part of?

A

CBC or hemogram

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Q

PBS is required for?

A

enumeration and checking of cellular elements

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Q

Microscopic examination aids in providing info regarding?

A

formed elements in blood

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Q

Reliability of information obtained depends on?

A

well-made and well-stained films

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Q

T/F: Smears must be prepared immediately as possible.

A

True

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Q

Blood sample

A

Anticoagulated EDTA blood

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Q

Stain used

A

Wright or Wright-Giemsa stain

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Q

Systematic approach:

A

10x
40x
OIO

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Q

Materials needd:

A

Glass slides, coverslips
Pasteur pipette
Anticoagulated blood
Tube holder
Pencil for labeling
PPEs
Canister with yellow bag
Sharps container
Oil cloth

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Q

Method 1

A

Two-Slide or Wedge method

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Q

Simplest and most common method for smear prep

A

Two-Slide or Wedge method

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Q

Procedures for Method 1

Size of drop ng blood

A

2-3mm in diameter

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Q

Procedures for Method 1

Drop of blood should be placed where?

A

1cm from the end

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Q

Procedures for Method 1

Fingers used to hold spreader slide

A

thumb and forefinger

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Q

Procedures for Method 1

Angle of spreader slide

A

30-45 degrees

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Q

Features of a Properly Made Blood Smear

The film must cover?

A

2/3 to 3/4 length

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Q

Features of a Properly Made Blood Smear

The film is what shape?

A

finger shaped (rounded at feather edge)

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Q

Features of a Properly Made Blood Smear

What edges should be visible?

A

lateral edges

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Q

Features of a Properly Made Blood Smear

The film is smooth without irregularities, like?

A

holes or streaks

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Q

Features of a Properly Made Blood Smear

When slide is held up to light, feathery edge should have a?

A

rainbow appearance

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Q

Portion observed in feathery edge

A

Zone of morphology

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Q

Unacceptable Peripheral Blood Smears and its Causes

A

chipped or rough edge
hesitation in forward motion
pushed too quickly
drop of blood was too small
not allowed to spread across the width of slide
dirt or grease
uneven pressure
time delay

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Q

T/F: Hematocrit levels can be attributed to smears.

A

True

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Q

Low hematocrit levels

A

Too thin

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High hematocrit levels

Too thick

Reminders once you have created the smear A good film includes a?

thick and thin portion, with gradual transition

Moving the pusher slide forward too slowly accentuates?

poor leukocyte distribution (pushing larger cells at very end of film)

The faster the film is air dried,

the better is the spreading

Slow drying causes?

formation of artifactual cell

Slide may be labeled on the frosted end or directly on the thicker end using?

lead pencil

The thickness of the film can be adjusted by?

changing the angle or speed of spreading

This can cause erroneous result in preparing the smear

high lipid content

Older technique that is used for making bone marrow aspirate smears.

Two-Coverslip or Two-Coverglass method

Method 2

Two-Coverslip or Two-Coverglass method

Recommended coverslips for Method 2

No. 1 or 1 1/2 cover glasses 22mm

Quality coverslip smear will appear similar to a?

thumb print

Making smears on coverslips require?

manual dexterity

Advantage of Method 2

- excellent leukocyte distribution - minimal amount of bone marrow is needed but many smears are made

Procedures for Method 2 Hold one coverglass on adjacent corners with?

Thumb and index finger

Procedures for Method 2 Blood must form a?

16-sided figure (letter B)

Procedures for Method 2 Before blood spreads, seperate coverglass by?

rapid, even, horizontal and lateral pull

Method 3

Spinner Method

This method would create a uniform blood film.

Spinner Method

Easy handling of wedge slide and uniform distribution of cells can be made with special types of centrifuges known as?

spinners

Spinner slide produces film in which?

all cells are separated (monolayer) and randomly distributed

Smears prepared using Method 3 have what that can be easily identified at any spot in the film?

white blood cells

On wedge smear, disproportions of ____ occur at tip of feather edge

monocytes

____ are seen just in from the feather edge

Neutrophils

___________ may occur at lateral edges of the films

Neutrophils and Monocytes

This is an automated slide making and staining system.

Sysmex SP-1000i

Automated Smear Preparation is dependent on?

hematocrit values

Film for automated smear prep is produced every ____ and with what?

30 seconds, printed label

Printed label for Automated Smear Prep contains?

- Patient name - Number - Date of preparation

This prepares slides automatically based on orders received from the LIS

DxH Slidemaker Stainer II Cellular Analysis System by Beckman Coulter

DxH Slidemaker Stainer II Cellular Analysis System by Beckman Coulter This creates and stains smears from?

- capped whole blood tubes - open-top tubes - pediatric whole blood tubes

DxH Slidemaker Stainer II Cellular Analysis System by Beckman Coulter How many slides can it create from a single 90uL sample?

4 slides

DxH Slidemaker Stainer II Cellular Analysis System by Beckman Coulter How many slides can it create from a single sample presentation?

12 slides

DxH Slidemaker Stainer II Cellular Analysis System by Beckman Coulter This creates multiple stain smear slides according to?

any parameter results

Wright stain is composed of?

- Eosin - Complex mixture of (Thiazines) including methylene blue, azure blue

This staining technique is certified by the Biological Stain Commission and is commercially available as solution or powder.

Staining Jar or Dip Method

T/F: Polychrome stains contain Eosin and Methylene blue.

True

Heparin causes?

blue background

Oxalate and Fluoride causes?

distortion of cell morphology

Reagents in staining dip or jar method is found in?

coplin jars

Solution 1

Methanol, 30 seconds

This will serve as fixative.

Methanol

Solution 2

Eosin, 6 seconds

Free eosin is acidic and stains basic components such as?

Hemoglobin or eosinophilic granules (red)

Solution 3

Methylene blue, 4 seconds

Free methylene blue is basic and stains acidic cellular components such as?

RNA (blue)

Solution 4

Buffer solution/aged distilled water, 45 seconds

Buffer solution

0.05M sodium phosphate (pH 6.4)

Aged distilled water

Placed in glass bottle for at least 24 hours; pH 6.4 to 6.8)

Slide must dry in a?

tilted position

Color of RBCs

pink to salmon

Color of Nuclei

dark blue to purple

Cytoplasmic granules of Neutrophils

lavender to lilac

Cytoplasmic granules of Basophils

dark blue to black

Cytoplasmic granules of Eosinophils

red to orange

Area between the cells should be?

colorless, clean, free of precipitated stain

Slides must be prepared within how many hours following blood collection?

2 to 3 hours

Oxidation on staining causes development of?

Formic acid

Other romanowsky-type stains:

- Giemsa - Leishman's - Jenner's - May-grunwald - MacNeal's