Blood Film Review

Haematology

Blood film review remains an indispensable diagnostic tool, despite advances in automated technologies. The blood film, or peripheral blood smear, allows for the direct visualisation of blood cells, providing insights that are often beyond the reach of automated analysers. Automated analysers, while efficient, can miss subtle morphological abnormalities such as dysplastic changes, reactive features, atypical cells, or early signs of malignancies. In some cases, the blood film review is the only reliable methodology to identify key disease associated markers. For instance, the presence of schistocytes in microangiopathic haemolytic anaemia can only be reliably identified through a manual smear review. Blood film review thus aids in the differential diagnosis of patients presenting with haematological abnormalities which cannot be detected through routine automated methods.

In addition to its role in patient diagnosis, blood film review plays an essential role in the detection of spurious FBC results caused by pre-analytical artefacts or sample handling. For example, the clumping of platelets in EDTA anticoagulated samples may lead to spurious reporting of thrombocytopaenia.

At Eurofins-Biomnis Ireland, blood film review is provided as part of the FBC package and is automatically requested based on FBC result criteria.

Preparation of patients: There is no physical preparation for the blood film review.

Precautions: Frozen, clotted, or grossly haemolysed samples cannot be analysed.

Sample Type:

Whole Blood K2/K3 EDTA anticoagulated or unstained blood smears prepared from such specimens.

Temperature: + 2-8ºC for whole blood. Unstained slides can also be stored at room temperature.

Miscellanous: N/A

Turnaround Time:

24 hrs

Sample Stability:

48 hours @ + 2-8ºC for whole blood.

8 weeks for blood smear slides prepared from whole blood EDTA anticoagulated specimens meeting the above stability criteria.

Instrument / Procedure:

Manual microscopy

Units:

Reference Range:

Precautions:

Rejection Criteria: Clotted sample, insufficient sample, grossly haemolysed sample, centrifuged specimens, unlabelled sample, mismatched patient ID, sample over 48 hours of collection, wrong sample tube (EDTA tube only).

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Accreditation Status:

Accredited

Non-Accredited

SAMPLE REQUIREMENTS FOR COAGULATION TESTS

PROCEDURE

Sample Requirements and Collection

Patients should be relaxed pre-venepuncture. Excessive stress and exercise will increase FVIII, vWF antigen and fibrinolysis. Venous occlusion should be avoided.
Difficult venepuncture with trauma may lead to platelet activation with release of PF4 from alpha granules.
Venous blood should be collected into coagulation tubes containing Sodium Citrate 3.2%, 0.105M.
Specimens must be mixed immediately post venepuncture to avoid clot activation, by GENTLY inverting the tubes 5 to 10 times.
The ratio of whole blood to anticoagulant is crucial to clotting times. A target blood to anticoagulant ratio of 9:1 is optimal. Under- or over- filled specimens will not be processed this can adversely affect results.
Any warfarin treatment should be mentioned on the request form.
Sample rejection Criteria: Clotted sample, grossly hemolyzed sample, underfilled/overfilled specimen, unlabeled sample, mismatched patient ID, aged samples, wrong sample tube (citrate tube only).

Transportation and Storage

PT/INR specimens should be transported to the laboratory at room temperature.
Coagulation specimens should ideally be analysed within 4 hours of collection. Where this is not possible, centrifuge specimens at room temperature (RT) @ 1500RCF for at least 15 minutes, and then carefully remove the plasma from the cells, transfer to a fresh plastic plain tube and freeze at -20oC.
Non-frozen coagulation specimens should be transported at RT ASAP to avoid deterioration of labile factors V and VIII.
Collection of blood through intravenous lines that have been previously flushed with heparin should be avoided. In the event blood is drawn from an indwelling catheter, the line should be flushed with 5ml of saline, and the first 5ml of blood or 6 times the line volume be drawn off and discarded before coagulation tube is filled.
Effect of freezing on Coagulation Specimens.
A 14days in-house study on the effect of freezing, on coagulation specimens at -20oC, showed that there was negligible and clinically non-significant effect of freezing on coagulation specimen results. Therefore frozen citrated coagulation samples are stable for 14 days at -20oC, post centrifugation. This study is available in-house for reference.

ESR Ref Ranges

Units of Measurement

MALE

FEMALE

>50 Years

mm/hr

0 - ≤12

0 - ≤15

<50 Years

mm/hr

0 - ≤8

0 - ≤10

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Analyte

Units of Measurement

MALE

FEMALE

WBC

10^9/L

4.0–10.0

4.0 - 10.0

RBC

10^12/L

5.0 ± 0.5

4.3 ± 0.5

g/dL

15.0 ± 2.0

13.5 ± 1.5

HCT

L/L

0.45 ± 0.05

0.41 ± 0.05

MCV

92 ± 9

MCH

29.5 ± 2.5

MCHC

g/dL

33.0 ± 1.5

PLT

10^9/L

280 ± 130

MPV

N/A

RDW

11.6 - 14.0

#Neut

10^9/L

2.0 – 7.0

(40 - 80%)

#Lymph

10^9/L

1.0 – 3.0

(20 - 40%)

#Mono

10^9/L

0.2 – 1.0

(2 - 10%)

#Eos

10^9/L

0.02 – 0.5

(1 - 6%)

#Baso

10^9/L

0.02 – 0.1

(<1 - 2%)

Analyte

Units of Measurement

MALE & FEMALE

WBC

10^9/L

Birth: 18 ± 8
Day 3: 15 ± 8
Day 7: 14 ± 8
Day 14: 14 ± 8
1 Month: 12 ± 7
2 Months: 10 ± 5
3–6 Months: 12 ± 6
1 Year: 11 ± 5
2–6 Years: 10 ± 5
6–12 Years: 9 ± 4

RBC

10^12/L

Birth: 6.0 ± 1.0
Day 3: 5.3 ± 1.3
Day 7: 5.1 ± 1.2
Day 14: 4.9 ± 1.3
1 Month: 4.2 ± 1.2
2 Months: 3.7 ± 0.6
3–6 Months: 4.7 ± 0.6
1 Year: 4.5 ± 0.6
2–6 Years: 4.6 ± 0.6
6–12 Years: 4.6 ± 0.6

g/dL

Birth: 18.0 ± 4.0
Day 3: 18.0 ± 3.0
Day 7: 17.5 ± 4.0
Day 14: 16.5 ± 4.0
1 Month: 14.0 ± 2.5
2 Months: 11.2± 1.8
3–6 Months: 12.6 ± 1.5
1 Year: 12.6 ± 1.5
2–6 Years: 12.5 ± 1.5
6–12 Years: 13.5 ± 2.0

HCT

L/L

Birth: 0.60 ± 0.15
Day 3: 0.56 ± 0.11
Day 7: 0.54 ± 0.12
Day 14: 0.51 ± 0.12
1 Month: 0.43 ± 0.10
2 Months: 0.35 ± 0.07
3–6 Months: 0.35 ± 0.08
1 Year: 0.34 ± 0.04
2–6 Years: 0.37 ± 0.03
6–12 Years: 0.40 ± 0.05

MCV

Birth: 110 ± 10
Day 3: 105 ± 13
Day 7: 107 ± 19
Day 14: 105 ± 19
1 Month: 104 ± 12
2 Months: 95 ± 8
3–6 Months: 76 ± 8
1 Year: 78 ± 6
2–6 Years: 81 ± 6
6–12 Years: 86 ± 9

MCH

Birth: 34 ± 3
Day 3: 34 ± 3
Day 7: 34 ± 3
Day 14: 34 ± 3
1 Month: 33 ± 3
2 Months: 30 ± 3
3–6 Months: 27 ± 3
1 Year: 27 ± 2
2–6 Years: 27 ± 3
6–12 Years: 29 ± 4

MCHC

g/dL

Birth: 33.0 ± 3.0
Day 3: 33.0 ± 4.0
Day 7: 33.0 ± 5.0
Day 14: 33.0 ± 5.0
1 Month: 33.0 ± 4.0
2 Months: 32.0 ± 3.5
3–6 Months: 33.0 ± 3.0
1 Year: 34.0 ± 2.0
2–6 Years: 34.0 ± 3.0
6–12 Years: 34.0 ± 3.0

PLT

10^9/L

Birth: 100 – 450
Day 3: 210 – 500
Day 7: 160 – 500
Day 14: 170 – 500
1 Month: 200 – 500
2 Months: 210 – 650
3–6 Months: 200 – 550
1 Year: 200 – 550
2–6 Years: 200 – 490
6–12 Years: 170 – 450

Reticulocytes

10^9/L

Birth: 120 – 400
Day 3: 50 – 350
Day 7: 50 – 100
Day 14: 50 - 100
1 Month: 20 – 60
2 Months: 30 – 50
3–6 Months: 40 – 100
1 Year: 30 – 100
2–6 Years: 30 – 100
6–12 Years: 30 – 100

#Neut

10^9/L

Birth: 4 – 14
Day 3: 3 – 5
Day 7: 3 – 6
Day 14: 3 – 7
1 Month: 3 – 9
2 Months: 1.0 – 5
3–6 Months: 1 – 6
1 Year: 1 – 7
2–6 Years: 1.5 – 8
6–12 Years: 2 – 8

#Lymph

10^9/L

Birth: 3 – 8
Day 3: 2 – 8
Day 7: 3 – 9
Day 14: 3 – 9
1 Month: 3 – 16
2 Months: 4 – 10
3–6 Months: 4 – 12
1 Year: 3.5 – 11
2–6 Years: 6 - 9
6–12 Years: 1 - 5

#Mono

10^9/L

Birth: 0.5 – 2.0
Day 3: 0.5 – 1.0
Day 7: 0.1 – 1.7
Day 14: 0.1 – 1.7
1 Month: 0.3 – 1.0
2 Months: 0.4 – 1.2
3–6 Months: 0.2 – 1.2
1 Year: 0.2 – 1.0
2–6 Years: 0.2 – 1.0
6–12 Years: 0.2 – 1.0

#Eos

10^9/L

Birth: 0.1 – 1.0
Day 3: 0.1 – 2.0
Day 7: 0.1 – 0.8
Day 14: 0.1 – 0.9
1 Month: 0.2 – 1.0
2 Months: 0.1 – 1.0
3–6 Months: 0.1 – 1.0
1 Year: 0.1 – 1.0
2–6 Years: 0.1 – 1.0
6–12 Years: 0.1 – 1.0

Reference Ranges:

Age

Absolute Reference Range

Age

% Reference Range

0 - 1 day

324 - 617 x10⁹/L

0 - 1 day

1.72 - 8.62%

1 - 5 days

85 - 400 x10⁹/L

1 - 5 days

1.9 - 9.1%

5 days - 1 mth

34.2 - 724 x10⁹/L

5 days - 1 mth

0.1 - 6.9%

1 - 3 mths

21.3 - 205 x10⁹/L

1 - 3 mths

0.1 - 6.27%

3 - 12 mths

8.0 - 171 x10⁹/L

3 - 12 mths

0.1 - 4.7%

1 - 3 yrs

55.6 - 120 x10⁹/L

1 - 3 yrs

0.35 - 2.95%

3 - 7yrs

16.4 - 120.7 x10⁹/L

3 - 7yrs

0.25 - 2.57%

Adult

35.2 - 122.8 x10⁹/L

Adult

0.75 - 2.7%

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