Transcript First time count

Quality control of Blood film
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When blood film needed?
 Blood count request:
 Is it a first time count or repeat count?
 First time count: Is it a routine screening test or special
category?
 If Routine: Analyzer report for blood count alone
 Film required if any flags are signaled
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First time count - If Special category, Film
required:
 Diagnosed blood disease patients
 Patients receiving radiotherapy and/or
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chemotherapy
Renal disease
Neonates
Intensive care unit
If special tests have also been requested for:
infectious mononucleosis, haemolytic anaemia, enzymopathy, abnormal haemoglobins
 If the clinical details on the request form indicate
lymphadenopathy, splenomegaly, jaundice or
suggest the possibility of leukaemia or lymphoma
 Specific requests by clinician
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Repeat count: Film required:
 Delta check positive when compared with previous record
 Any flag occurs in present count
 On each occasion for patients with known blood diseases, for
neonates, and when specifically requested by clinicians
International Society for Laboratory Hematology
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FLAGGING OF AUTOMATED BLOOD COUNTS
 "Flagging" refers to a signal that the specimen being
analyzed may have a significant abnormality because one or
more of the blood count variables are outside specified limits
(usually 2SD) or there is a qualitative abnormality that
requires a quality control check and/or additional
investigation.
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Flag4
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Making blood film
 Blood film can be prepared from fresh blood without
anticoagulant or from EDTA anticoagulanted blood.
 blood film should be made on clean glass .
 Clear without any dust
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Making blood films
Three basic steps to make blood film:
1.Preparation of blood smear.
2.Fixation of blood smear.
3.Staining of blood smear.
Optimal blood smear characteristic
 minimum 2.5 cm in length terminating at
least 1 cm from the end of the slide
 Gradual Transition in thickness from thick to thin area ending
in a Square or straight edge
 No streaks , waves , or troughs
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Romanowsky stain
 Romanowsky stain→Eosin Y and Azure B)
Eosin:Acidic Dye bind to Basic groups
(Hb,Granules)
→ reddish or orange color
Azure B: Dye bind to nucleic acid & nucleoproteins →Blueviolet color
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Good vs. Bad
Smears
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Good Peripheral Blood Smear
Prepare blood films within 4(3) h of the blood collection in K EDTA.
Stain the film within one hour of preparation with a Romanowsky stain,
containing fixatives; or fix within one hour with "water-free" (i.e., <3%
water) methanol for later staining.
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Peripheral Blood Smear
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The thickness of the spread notes
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Sources of error in preparation of a blood smear
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problem
Resolution
Presence of crenated
erythrocyte
Dry smear quickly and thoroughly
Thin smear due to anemia
Increased spreader slide angle and
increased push speed
Thick smear due to polycythemia
Decrease spreader slide angle and
decrease push speed
Presence of agglutinated
erythrocytes associated with cold
agglutinin disease
Warm blood in 37°C for 15 min
prior to preparing smear
Increased viscosity associated with
multiple myeloma
Decrease spreader slide angle and
decrease push speed
Characteristic of a properly stained blood
smear
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Type of evaluation
Characteristic
Macroscopic
Smear is pinkish purple in color
Microscopic
Blood cells are evenly distributed
Areas between cells are clear
Erythrocytes are orange red
Neutrophilic granules are lilac
Eosinophilic granules are red
orange
Lymphocytes cytoplasm is blue
Leukocytes nuclei are purple
Precipitated stain is minimal or
absent
problem
Potential causes
Excessively blue or dark stain Prolonged staining
Inadequate washing
Too high an alkalinity of stain and /
or buffer
Thick blood smear
Excessively pink or light stain Insufficient staining
Prolonged washing
Too high an acidity of stain and / or
buffer
Presence of precipitate
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Unclean slides
Drying during staining process
Inadequate filtration of stain
problem
Potential causes
Pale stainig
Old staining solution
overused staining solution
Impure dyes
High ambient temperature
Blue Background
Prolonged storage before fixation
Blood collection into heparin as
anticoagulant
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‫رنگ پس از تهيه از نظر آلودگی قارچی و ميکروبی وهر گونه رسوب و پارتيکل وهمچنين‬
‫نحوه رنگ گرفتن سلول های خونی بررس ی می گردد‪.‬رنگ آميزی گسترش های خونی در‬
‫هر ران کاری موارد فوق بررس ی می گردد که بصورت مکتوب ومستند بايد در‬
‫آزمایشگاه قرار گيرد‪.‬کيفيت رنگ آميزی مورد قبول سلول ها مطابق جدول زير می‬
‫باشد ‪.‬‬
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Leukocyte Differential Count
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 How can you estimate the WBC from
PBS?
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Estimation of WBC
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Corrected WBC
Nrbc/100 wbc
If2810 (5) or more Nrbc are observed , corrected WBC must be calculated
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How can you estimate the PLT from
PBS?
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Estimation of PLT
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DIFFERENCES BETWEEN
CAPILLARY & VENOUS BLOOD
 The differences may be exaggerated by cold with resulting slow capillary
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blood flow.
The PCV, RBC, and Hb of capillary blood are slightly greater than in
venous blood.
The WBC & neutrophil counts are higher by about 8%;
The monocyte count is higher by about 12%, and in some cases by as
much as 100%, especially in children.
Conversely, the platelet count appears to be higher in venous than in
capillary blood; this is on average by about 9% and in some cases by as much
as 32%.
This may be the result of adhesion of platelets to the site of the skin
puncture.
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‫نحوه گزارش الم خون محیطی‬
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Grading scale
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1(+)
2(++)
1-6 per oil
imm. field
7-10 per
OIF
3(+++)
4(++++)
11-20 per OIF > 20 per OIF
Hypochromia (correlate with MCHC)
 1+ :area of central pallor is
½ of cell diameter
 2+ : area of central pallor is 2/3 of cell diameter
 3+ : area of central pallor is
¾ of cell diameter
 4+ : thin rim of hemoglobin
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Quality control of BG&Rh
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‫‪ ‬انجام گروه خونی به دو روش ‪:‬‬
‫‪ Cell Type‬‬
‫‪ Back Type‬‬
‫‪ ‬توسط ‪ 2‬نفر‬
‫‪ ‬انجام تست ‪ Du‬جهت گروه های خونی‪Rh (-) :‬‬
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crenation
water artifact rbc
stain precipitate
smudge cells leukemia
Shift to left wbc
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dohle body
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AML M1
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AML M2
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AML M3
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AML M4
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AML M5
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‫پالسماسیتوئید سل‬
Megathrombocyte, Normal capillary blood
smear
Epithelial cell, microorganisms
Late Neutrophil Surrounded - Platelets, Platelet
Satellitism
Mature Neutrophils surrounded by Platelets,
Satellitism
Acute Megakaryocytic Leukemia (M-7) in
Remission
Large lymphocyte with nucleolus
hypersegmented neutrophil is present along with
macro-ovalocytes in a case of pernicious anemia
The nucleated RBC in the center contains
basophilic stippling of the cytoplasm
peripheral blood smear demonstrates many
larger bluish reticulocytes as well as smaller
RBC's lacking central pallor--spherocytes
This peripheral blood smear demonstrates
several ring forms of Malaria (Plasmodium
vivax) in the red blood cells.
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Howell-Jolly bodies, or inclusions of nuclear
chromatin remnants