HEMA 2 LAB Flashcards by Buenavista, Leira Stela

His mother experienced 3 weeks of bleeding following a dental
extraction procedure. No remarks made about his father.

Look closer to laboratory work out

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Laboratory Investigations show:
Hgb - 15 g/dL
Hct - 44%
Platelet count - 245, 000/uL
Bleeding Time - 10 minutes ( Duke Technique) - Abnormal
Protime - 12 seconds
APTT - 55 seconds (Done Twice) - Abnormal

Check Platelet Aggregation

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He did the platelet aggregation study. The result was abnormal in ADP, EPINEPHRINE and COLLAGEN. ANd it shows there is
abnormal aggregation in RISTOCETIN.

Bernard Soulier Syndrome

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The automated CBC and platelet reading didn’t display any flags for abnormal cells. Does this mean all the morphology of the cell is normal?

I thought it is Bernard-Soulier Dse because of the abnormal platelet aggregation in ristocetin. If this is BSD, the APTT should be normal and there should be flags of large platelets in the automated method, which is not present in this case. Therefore, I conclude that this is Von Willebrand

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What is the purpose of Ristocetin aggregation assay?

Ristocetin-induced platelet aggregation (RIPA) is used as an in vitro test to determine the presence and integrity of the platelet glycoprotein Ib and von Willebrand factor interaction and is usually performed using platelet-rich plasma (PRP). It can be used to differentiate Glanzmann Thrombasthenia from Bernard-Soulier Dse and VWF Dse.

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There are several methods used to test the bleeding time. Between Duke and Ivy Method which one is preferred?

I prefer using the ivy method because this is more reliable. The intercapillary pressure is standardized therefore, we can reduce errors and make sure the scouts are screened properly

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How can we maintain the intercapillary pressure in Ivy Method?

Increase the cuff pressure and hold the pressure exactly 40mmHg for the entire period

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Next step is to make separate cuts in the forearm. What is the
recommended depth of the wound?

3mm deep will be enough (Adults)
1.6mm (Infants/newborns)

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After making the incision what is the next thing to do?

Start the timer immediately and wick the blood using filter paper every half minute

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How do you record the bleeding time?

Count the number of blot multiplied by 30 (seconds)

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What is the normal bleeding time (IVY Method)

If the bleeding time stops 2-9 minutes

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What is the normal bleeding time (Duke’s Method)

1-5 minutes (normal range)

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Bleeding time assess which parameters?

Bleeding time assesses platelet functional integrity and platelet number.

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Sasha experiencing weakness and fatigue. She also experienced bleeding gums while brushing her teeth. Her mom experiences nosebleed once a month and her dad experienced prolonged bleeding 3 years ago when he accidentally cut himself with a hunting knife. What laboratory test should we do to diagnose a disorder?

APTT, PT, Bleeding Time, Platelet Aggregation, RIPA
and Platelet Count (Best Choice kay all of those methods
assist hemostasis)

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APTT, PT, Bleeding Time, Platelet Aggregation, RIPA and Platelet Count (Best Choice kay all of those methods assist hemostasis)

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Bleeding Time and Platelet Count, aggregation and RIPA (ristocetin induced platelet aggregation)

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Prothrombin Time, APTT

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Here are the laboratory results:
APTT- normal
PT- normal
Bleeding time - prolonged
Platelet aggregation - abnormal
RIPA - normal (positive)
Platelet COunt - borderline normal
What can you conclude about these results?

This Glanzmann’s Thrombasthenia since her results in RIPA (positive), APTT and PT are all normal but with prolonged bleeding time

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To do a Direct Platelet Count using Rees Ecker Method
What is the color of the tube this test requires?

EDTA (purple top) for platelet counting

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What is the best site for extraction of blood?

Median Cubital Vein

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What pipette do we use in the Rees Ecker Method?

RBC Pipette

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In routine where do we aspirate nga mark?

1 mark and 101 diluting fluid

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In what instances can we lower aspiration of the sample?

When the platelet is increased

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At what level of platelet concentration count can we use the WBC
pipette?

Less than 50, 000/mL

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What is the purpose of rinsing the RBC pipette with Reese Ecker diluting fluid?

Aside from being diluent, this fluid stains the platelet so we can count them easily in the Light Microscope

What is the purpose of rinsing the RBC pipette with Reese Ecker diluting fluid? ( answer: Aside from being diluent, this fluid stains the platelet so we can count them easily in the Light Microscope) What if we use a Phase contrast microscope? What method is being used?

Brecher-conkrite method Diluting fluid of Brecher-Conkrite- ammonium oxalate (no stain)

What mark can we draw blood?

Draw until 0.5 mark and use 200 as the dilution factor

How many minutes are we going to put this on the pipet shaker?

5-10 minutes

We count platelets using 40x magnification on which area?

Central Large Square containing 25 smaller squares

On the first square. I counted a total of 237 platelets and on the other square I counted a total of 210. Is this alright?

Not acceptable. Do it Again (27 difference)

Here’s my new platelet count 227 and 215

Since the difference is less than 10% those values are acceptable. Formula: Platelets/uL (227+215) 𝑥 200 / 0.1

442, 000 platelets/uL

Slight increase

What is the component present in cell that’s why ga positive in Periodic acid-schiff (PAS)?

Carbohydrate

If we inhibit the non-specific esterase may sodium fluoride

Nd ma stain

Other name for eosinophil depression test

Thorn’s Test

Reagent used to lysed WBC except eosinophil

Sodium Carbonate

Basophil will distinguish from other cells using

Toluidine Blue Stain under Toluidine Blue method/Cooper and Cruickshank Method

Important stain for eosinophil count

Phloxine

Important stain in Basophil count

Toluidine blue (appear metachromatic purplish red/reddish purple)

Complete Component

Pilot Solution (phloxine, propylene glycol, sodium carbonate, heparin)

What is this substance to lyse the RBC but the basophilic granules become insoluble?

Cetylpyridinium chloride

Terminal deoxynucleotidyl transferase (TDT) uses to stain

Polymerase found in lymphoblast nuclei

What is the relationship between Eosinophil count and level of corticosteroids

- In hyperadrenalism cushing's disease eosinophil will decreased while hypoadrenalism eosinophil will increased - INVERSELY PROPORTIONAL

In Nitroblue Tetrazolium Test (NBT) colorless

Stain Neutrophil (when mixed in neutrophil it becomes blue-black formazan precipitate) especially the granules

In Other cytochemical staining what enzyme is present?

Isoenzyme 5 of acid phosphatase (that is why hairy cells are TRAP positive)

Isoenzyme 5 acid phosphatase

Tartrate resistance present in hairy cells

LAP SCORE

- Increased LAP - Leukemoid Reaction - Decreased LAP - CGL/CML

With regards to cytochemical stains. What are the 3 cytochemical stains that are positive in AML but negative in ALL

- MPO - SBB - Specific Esterase

How many platelets are seen per 100 RBC

3-10

How many platelets are seen per OIO?

7-21

Percent of Platelet:

Circulation- ⅔ 75% Spleen - ⅓ 25%

How many platelets are produced per megakaryocyte?

2000-4000

Approximately in healthy adult women. What is the platelet concentration? (total blood volume)

1 trillion

Per cu/mL kabilogan nga blood volume

250 million

Indirect method of platelet counting. What factor used?

2, 000

In an automated analyzer. What are the situations that can falsely increase/elevated platelet count?

- Fragmented RBC/schistocytes - Cytoplasmic fragments of WBC

In an automated analyzer. What are the situations that can falsely decrease platelet count?

- Old specimen - Giant platelets - Satellitism - Clumping/aggregates of platelet

(Module 3) Simplate Method

Contains a spring-loaded blade within a plastic case which holds a double blade.

(Module 3) Surgicutt Method

Utilizes a slicing action using a surgical blade

(Module 3) Mielke Method

Utilizes a template containing a standardized slit in place of a disposable lancet

In Rumpel-Leede/ Capillary Fragility Test/Hess Test/Tourniquet Test if positive there’s a presence of?

Petechiae

Platelet adhesiveness test. If we collect the sample using a glass bead system the expected platelet count will be?

Decreased compared to the original sample collected using EDTA

Conditions that causes RBC’s Fall out

- Glanzmann thrombasthenia - Disseminated Intravascular coagulation (DIC) - Hypofibrinogenemia - Dysfibrinogenemia

(Module 3) Start of clot retraction

- 30 mins

(Module 3) Normal Clot retraction

2-4 hours - Normal 4-24 hours - Poor retraction Above 24 hours - there is no retraction at all

In what method do we use castor oil?

Hirschboeck Method

How do we report clot retraction?

-using the grading or percentage of the serum extruded ○ 0: no serum extruded ○ 1+: 5-10% serum extruded ○ 2+: 10-20% serum extruded ○ 3+: 20-35% serum extruded ○ 4+: 35-50% serum extruded

Qualitative abnormalities of platelets are suspected if:

- Bleeding manifestation (mucocutaneous bleeding) - Platelet count above 50, 000/mL

3 Methods of Aggregation

● Optical density/light transmittance - we are using platelet rich plasma ● Electrical impedance/ Electrical resistance - we are using WHOLE BLOOD ● Lumi Aggregometry- Either Platelet rich plasma/ Whole blood

Temperature during aggregometry

18-24 degree celsius

How long would you analyze after centrifugation to obtain platelet rich plasma?

Within 30mins

What reagent are used in Lumiaggregometry?

Firefly-derived luciferin-luciferase reagent

Formula of %platelet adhesiveness

(𝑃𝐶 𝑤𝑖𝑡ℎ𝑜𝑢𝑡 𝑔𝑙𝑎𝑠𝑠 𝑏𝑒𝑎𝑑𝑠 − 𝑃𝐶 𝑤𝑖𝑡ℎ 𝑔𝑙𝑎𝑠𝑠 𝑏𝑒𝑎𝑑𝑠 / 𝑃𝐶 𝑤𝑖𝑡ℎ𝑜𝑢𝑡 𝐺𝑙𝑎𝑠𝑠 𝑏𝑒𝑎𝑑𝑠) x 100

What is the relationship between clot retraction and platelet concentration?

Directly Proportional

Anticoagulant of choice in Aggregometry test

3.2% Sodium Citrate

When do we start the timer in bleeding time?

As we seen the blood

HEMA 2 LAB Flashcards

(76 cards)