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Ola During Children’s / Princess Christian Maternity Hospitals – Abertawe BroMorgannwg University Health Board Link

CLINICAL LABORATORY SERVICES DEVELOPMENT PROJECT

OLA DURING/PCMH LABORATORY, SIERRA LEONE – ABMU HEALTH BOARD, SWANSEA UK LINK

Standard Operating Procedure: Using an adjustable-‐volume pipette

Version: #01; January 12th, 2011.

This Standard Operating Procedure (SOP) has been developed by the Ola During Children’s / Princess Christian Maternity Hospitals, Freetown, Sierra Leone – Abertawe BroMorgannwg Health Board, Swansea, UK Link. The Clinical Laboratory Services Development Project was funded by the International Health Links Funding Scheme supported by the British Council and Department for International Development, UK. This SOP was developed by:

• Borbor Sam Aliyah, Mohammed Mansaray and Umar Kamara, Ola During Children’s Hospital, Freetown, Sierra Leone and

• Angela Allen, Stephanie Allen and Stephen Allen, Abertawe BroMorgannwg

University Health Board, Swansea, UK. This SOP is one of a series of resources developed to support clinical laboratories in low resource settings. These resources are available from the Ibadan-‐Swansea Partnership website: see http://isp.swanih.org/ We hope that this SOP will be helpful in ensuring the quality of laboratory practice. The information is based on the manufacturer’s instructions and, to our knowledge, is correct. However, please note that the responsibility for the use of this SOP rests with individual laboratories and their staff. We are keen to further develop our resources so that they are as helpful as possible. Please do let us know of any comments, suggestions or feedback from using these resources. Please contact Angela Allen: [email protected]

Authorised signature: Number: Date:

Standard Operating Procedure: Using an adjustable-‐volume pipette



Why is this important? The measurement and delivery of the correct volumes of reagents, controls, standards and samples ensures accuracy of laboratory tests and results. Step 1: Choose the appropriate pipette

SL 20 Measures 2-‐20μl

Tens of μl

μl

Decimals

SL 200 Measures 20-‐200 μl

Hundreds of μl

Tens of μl

μl

SL 1000 Measures 100-‐1000 μl (0.1-‐1ml)

Thousands of μl

(ml)

Hundreds of μl

Tens of μl

Pipette set at 20.0 μl

Pipette set at 200 μl

Pipette set at 1000 μl (1 ml)

Remember! 1000μl = 1 ml



Step 2: Adjust pipette volume Step 2.1: If the pipette has a lock, unlock.

Step 2.2: Set pipette to required volume by turning the plunger

• Turn plunger anti-‐clockwise to increase volume

• Turn plunger clockwise to decrease

volume

Step 2.3: After setting the volume, lock (if pipette has a lock)

Lock

Plunger



1ml 200μl 20μl 1ml 200μl or 20μl Step 3: Attach suitable tip Step 3.1: Choose the right-‐sized tip for your pipette

Step 3.2: Place tip firmly on end of the pipette

Step 4: Prime pipette before each use Step 4.1: Push plunger down to the first stop to expel air from tip

Step 4.2: With plunger still depressed, insert tip into reagent or sample



Step 4.3: Slowly release plunger to take up liquid

Step 4.4: Check that there are no air bubbles

Step 4.5: Push plunger all the way down to expel reagent or liquid and then release plunger

Your pipette is now primed. The inside of the tip has been coated with reagent or sample. It is now ready to measure an accurate volume.

Air bubble



Step 5: Taking-‐up the reagent or sample Step 5.1: Push plunger down to the first stop

Step 5.2: Place tip in reagent or sample. Release plunger.

Step 5.3: Check that there are no air bubbles. Wipe the pipette tip with tissue to remove excess liquid.

Step 5.4: Push plunger all the way down to release reagent or sample.



Finally, if it is a viscous reagent or sample, push and release the plunger a few times (“flushing”) to make sure that all of the reagent or sample has been dispensed. This is illustrated with a blood sample.

Some of blood sample

remains in tip

All of sample dispensed after flushing

Step 5.5: Remove tip and discard appropriately. Do not discard tips into plastic bags as the tip can puncture that plastic causing a potential health hazard.



General care of pipettes Wipe the barrel of the pipette with absorbent towel soaked in 70% alcohol after use.

• When not in use, store in a safe place • Never leave a used tip on the end of the

pipette

• Never leave your pipette protruding from the work bench

Look after your pipettes well and they will give you years of reliable service. Make sure that every pipette is serviced and calibrated by an engineer regularly (usually once a year).

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