manual procedure of hematology department

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manual procedure of

HEMATOLOGY DEPARTMENT

INTRODUCTION:

The department of hematology caters to a vast majority of patients population both outpatients and in-patients.

It is supervised by specialist in laboratory hematology and coordinated by trained and competent medical technologist.

ROUTINE TEST DONE:

1. Complete Blood Count (CBC) including hemogram, differential count and platelet count

2. ESR

3. Blood grouping and typing

4. Bleeding time

5. Clotting time

6. Blood film for malaria

7. Peripheral film examination

8. Reticulocyte count

9. Sickling test

10. Prothrombin time (PT)

11. Activated partial thromboplastin time (APTT)12. FDP
13. Hemoglobin alkaline electrophoresis

EQUIPMENTS USED:

1. BLOOD CELL ANALYZER:

· SYSMEX SE-9500

· COULTER – JT

· COULTER – T 540

· COULTER HMX

2. DIAGNOSTICA STAGO (ST4) COAGULOMETER

3. HELENA ELECTROPHORESIS EQUIPMENT

DUTIES AND RESPONSIBILITIES OF
LABORATORY TECHNICIAN

HEMATOLOGY DEPARTMENT

I – Observing General Work Etiquette:

· Technicians must be alert, pleasant in behavior, and must carry out the prescribed work with full interest and enthusiasm. They must totally get involved in the work matters and create a team work atmosphere in running the show.

II – Receiving and processing of samples:

· Checking the correct sample by noting the patient’s identification details and the tests requested.

· Entering the patient’s I.D. in the logbook.

· Looking for any discrepancy in sample I.D., inadequate volume, blood clots, etc. and ordering fresh samples.

· Running the samples and entering the results in the logbook.

· Signing the reports, putting name stamps before dispatching.

· The undersigned will be responsible for the authenticity of the results.

III – Daily, weekly & monthly maintenance of the machines:

· Running daily controls and checking data.

· Maintaining quality control programme.

IV – Any discrepancy or abnormality in the results noted should be brought to the notice to the head of the department for corrective measures.

STANDARD OPERATING PROCEDURE

FOR
COULTER T 540

1. Check for the adequate volume of consumables: DILUENT, LYSE, and CLEANING AGENT.

2. Switch ON the machine.

3. Check the manometer pressure and adjust it to 30 PSI and 5 PSI respectively by using the knobs so as to have good vacuum during machine operation.

4. Adjust the date and enter the correct date. Press ENTER until SELECT FUNCTION appears.

5. Press START-UP.

6. Look for the background results by taking the print out or by pressing individual button for values present on the machine.

7. If background results are ok, run control sample.

8. If the control values are within the range, proceed to sample analysis.

TROUBLE-SHOOTING GUIDE:

1. If the screen shows low vacuum message, adjust the pressure to 30 PSI and 5 PSI respectively and then press RES 1 & RES 2 simultaneously.

2. If the screen shows CHANGE REAGENT message, look for the amount of diluent present. Replace it if it is low then press F14 and then do START-UP until the background results are within the range.

3. If the diluent is enough but the screen shows LOW REAGENT MESSAGE, press F14 and then do START-UP until the background results are within the range.

· Run controls if background results are ok.

· If controls are within range, proceed to sample analysis.

4. For any problem during machine run, contact the BIOMEDICAL Engineer.

BASIC PROCEDURE FOR RUNNING

COULTER- JT BLOOD CELL COUNTER

1. Swich ON machine/monitor.

2. Enter the date then press ENTER twice until SELECT FUNCTION appears.

3. Press START-UP button and look for the background results by pressing keys on the monitor screen (3-ENTER, 1-ENTER, 1- ENTER). If the background results is NOT OK, do START-UP again until it is OK. Press RETURN twice.

4. Run controls if background is OK, Check results by pressing the keys on the monitor screen (4- ENTER, 1-ENTER, 1- ENTER). If the controls are OK, get the sample analysis mode on the monitor screen by pressing return thrice then 1-ENTER, 1- ENTER.

TO CHANGE REAGENT, DILUENT and LYSE:

· Press F16 – to prime diluent

· Press F17 – to prime lyse

· Press F18 – to prime cleaning agent

TO SWITCH OFF THE MACHINE / MONITOR:

1. Select Function 02. ENTER.

2. Do this twice.

3. Press the WHOLE BLOOD button.

4. When the NEXT TEST appears, switch off the machine/ monitor.

BASIC PROCEDURES FOR RUNNING CELL COUNTER
HMX

1. Switch ON the power button at the back of the machine.

2. Switch ON the reset button at the left corner of the machine.

3. Wait till the access screen appears on the monitor.

4. Do start-up procedure daily before running the samples.

5. Press F9 and go to Diluter Function.

6. Select Start Up from the Diluter Function and Enter.

7. If the background results are OK, proceed to run controls. If any problem exist, see Trouble Shooting Guide.

8. Procedure for Control Run :

· Select F2 and Enter.

· Look for Control File.

· Select the desired control runs (Normal, Abnormal I & II) and press Enter.

9. Run the desired controls and look for the values. If it is OK, proceed to run samples. If any problems, see Trouble Shooting Guide.

SAMPLE ANALYSIS:

There are two modes present : Primary & Secondary

Samples_on Primary Mode :
1. Quantity of the samples must be more than 1 ml.
2. Select Function should appear on the lower right of the screen.
3. Select F1 then F2 twice and select Primary Mode then Enter.
4. Sample ID :
· Look for Work list on the screen by pressing F1 and
enter cassette ID No. & Patient ID No.
5. Take out the cassettes and put the samples and keep it in the loading place. The machine will automatically run the samples and print the results.

Samples_in Secondary Mode :
1. Select F1 & F3 twice and select Secondary Mode.
2. Enter the ID No. on the screen.
3. Open the sample & immerse the aspirator tip on the sample.
4. Press and release the sample bar then remove the sample when you hear the beep.

CLOSING THE MACHINE :
1. Look for Select Function
2. Select Diluter Function, SHUTDOWN
3. Press ENTER to begin
4. Close the machine by switching OFF the main button

PROCEDURE FOR RUNNING

SYSMEX SE – 9500:

1. Turn ON the power starting from power supply, compressor, main unit, DMS and turn OFF in the reverse order.
2. Check the pressure. It should be 0.2 – 0.3 and >0.04 in the other gauge.
3. Set the button CBC + DIFF.COUNT ON , then ENTER.
4. The machine will do auto rinse 2-3 times to get a clear background. If background is not ok, do auto rinse again.
5. For manual mode: SELECT ID #, ENTER.
6. Feed the sample be pressing the green button and wait for the beep. Release sample after the beep.
7. Result will be printed automatically.
CONTROL:
1. Press F4, look for QC menu.
2. Press 2 from small window, select the file:
· File I – Normal
· File II – Low
· File III – High
3. Feed the control. Look for +/- sign (+: above range; -: below range).
CALIBRATION:
1. Press shift and F7 at once, Look for change 3.
2. Press 4 for calibration.
SHUT DOWN:
1. Select from main menu.
2. Press shutdown.
3. Place the clean cell in the aspirator
TROUBLE SHOOTING:
· If background results are not ok, try autorinse 2 times.
· If not ok, press clog removal or F10 for operator guide.
· If still not ok, open the main door and press the clear aperture button.
· If problem still exists, see operator guide book or contact biomedical engineer.

TEST FOR SICKLE CELLS

PRINCIPLE:
One drop of blood is mixed with one drop of sodium metabisulfite reagent on a slide. If the red cells contain an abnormal haemoglobin called haemoglobin S, they will become sickle-shaped or half-moon shaped.

The reagent removes oxygen from the cells, allowing sickling to take place.

Haemoglobin S is an inherited abnormal haemoglobin. If inherited from both parents it causes sickle-cell anaemia, a serious disease. If inherited from only one parent it causes sickle-cell trait, which does not usually cause disease.

The sickle-cell slide test does not distinguish between sickle-cell anaemia and sickle-cell trait.

Haemoglobin S occurs mainly in tropical Africa but also in the Middle East and among American Negroes.

MATERIALS:
· Glass slide and coverglass
· Pasteur pipette (or dropping pipette)
· 20 g/l sodium metabisulfite- fresh

PROCEDURE TO PREPARE 2% SODIUM METABISULFITE:
1. Weigh 0.5 g of sodium metabisulfite reagent.
2. Using a graduated cylinder, measure 25 ml distilled water.
3. Mix thoroughly.
4. Transfer to a clean amber bottle and cover with parafilm.
NOTE: Daily prepare fresh 2% sodium metabisulfite solution.
· Container to prevent drying of the preparation, such as a petri dish.

SAMPLE COLLECTION:
· EDTA anticoagulated venous whole blood

PROCEDURE:
1. Place a small drop of EDTA blood (about 0.02 ml) in the center of a slide.
2. Add an equal drop of the sodium metabisulfite solution.
3. Mix carefully with the corner of the coverslip. Cover with the coverslip, making sure that no air bubbles form.
4. Place in a petri dish that has wet filter paper in the bottom. Support the slide in two sticks.
Note: When using a reducing reagent such as sodium metabisulfite it is not necessary to seal the preparation.
5. Wait 1 hour.
6. Examine under the microscope (x40 objective) after 1 hour.
If the result is negative, re-examine after a further overnight incubation.

RESULTS:
NEGATIVE RESULT: the red cells remain round.
POSITIVE RESULT: the red cell become sickle-shaped or banana-shaped and often with spikes.

· It is important to examine several parts of the preparation, as sickling can occur more quickly in one part than in another.
· Do not mistake for sickle-cells cells lying on their side or crenated cells.
· ALL POSITIVE and INDETERMINATE CASES SHOULD BE CONFIRMED BY HEMOGLOBIN ELECTROPHORESIS…solution.

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manual procedure of hematology department 70588

manual procedure of hematology department 54828

manual procedure of hematology department 409347

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