Repeater Pipette – Parts, Procedure, uses

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Repeater pipette is a special type of pipette used to dispense the same volume of liquid many times after a single aspiration. It is also called stepper pipette or multi dispenser pipette. It is used when repetitive dispensing is required in routine laboratory works.

It works on positive displacement mechanism in which a piston inside a disposable syringe type tip comes in direct contact with the liquid. There is no air cushion like air displacement pipette. Due to this direct contact, better volume accuracy is obtained and difficult liquids can be handled.

It is suitable for viscous, volatile, dense liquids and liquids that form foam. It also reduces aerosol formation during dispensing, so it is safer for infectious, toxic and radioactive samples. Repeater pipettes are available as manual and electronic models and it improves speed in high throughput tasks like filling microplates and aliquoting reagents.

It also reduces thumb force and reduces repetitive strain injury (RSI) during repeated pipetting. The same aspiration can be used for multiple equal dispenses and it saves time and reagent handling errors.

How Repeater Pipettes Work

Repeater pipette works on the principle of positive displacement. In this, the piston makes a direct tight contact with the liquid inside a special disposable syringe tip. The air cushion present in normal air displacement pipette is not there, so the volume delivery is more consistent.

In working, a large volume of liquid is aspirated only once into the tip. After this, the piston is moved down in equal measured steps and the same volume is dispensed repeatedly as many aliquots. In manual repeater pipette, this step movement is controlled by internal ratchet and gear system. In electronic repeater pipette, the step movement is controlled by a microprocessor and stepper motor.

For accuracy, a reverse stroke is usually done in the beginning and the first drop is discarded. This helps to settle the mechanism and to remove any trapped air bubbles. At the end, a residual stroke lock is present which keeps a small leftover liquid and prevents an inaccurate final dispense volume.

Parts of Repeater Pipettes

  1. Volume selection wheel / dial. It is a dial used to set the dispensing increment and select the volume to be dispensed in each step.
  2. Filling lever. It is used to retract the piston and aspirate the liquid into the syringe tip.
  3. Dispensing / operating lever. It is used to advance the piston for dispensing equal aliquots. In many models it also works with tip ejection function.
  4. Tip sensor. It is a mechanical or electronic sensor that reads the coding of the attached syringe tip. It helps to identify tip size and adjust volume calculation.
  5. Piston locking mechanism. It is an internal locking part that connects and secures the tip piston with the instrument. It helps to avoid mechanical backlash and gives direct movement.
  6. Step counter / digital display. It is a display that shows selected volume and number of dispensing steps during the series.
  7. Hand rest / finger hook. It is a curved support where index finger rests. It reduces gripping force and decreases hand strain during repeated work.
  8. Tip ejection system. It includes ejector button or lever for removing syringe tips without touching. It helps in safe disposal and avoids contamination.
  9. Syringe style tip (consumable). It is the disposable tip used in repeater pipette. It has an outer plastic tube, internal piston, seal, and coding crown for recognition. In some electronic models a center sleeve is also present for stable connection.

How to Use Repeater Pipettes?

  1. Select and attach the tip. The correct disposable syringe tip (Combitip) is selected based on required volume. The filling lever is pushed fully down and the tip is inserted into the pipette until it clicks and fits properly. For 25 mL or 50 mL tips, a reusable adapter is attached first.
  2. Set the volume. The volume selection dial is adjusted to the volume to be dispensed for each aliquot. The setting is checked before aspiration.
  3. Aspirate the liquid. The pipette is held vertically and the tip is immersed about 2 to 3 mm below the liquid surface. The filling lever is pulled up slowly to aspirate the liquid. Sudden pulling is avoided to prevent air bubble formation.
  4. Reverse stroke (discard step). The dispensing lever is pressed once and the first dispensed amount is discarded into the original container or waste tube. This step settles the internal mechanism and removes trapped air bubbles.
  5. Dispense the aliquots. The tip is placed against the inner wall of receiving container at about 30 to 45 degree angle. The dispensing lever is pressed to release the set volume. The same press is repeated for each tube or well to deliver equal volume.
  6. Empty the residual liquid. After dispensing series is completed, the filling lever is pushed to the bottom to expel the leftover liquid into source vessel or waste container.
  7. Eject the tip. The tip ejector keys are pressed or the bottom and top levers are pushed together (depending on model). The used tip is dropped into disposal bin without touching by hand.

Use of Repeater Pipettes

  • Aliquoting samples and reagents. It is used to divide a large volume into many small equal portions. It helps in storing samples like serum, proteins and antibiotics and reduces repeated freeze thaw damage.
  • High throughput vessel filling. It is used for rapid dispensing of same volume into many tubes and microplates (96 well, 384 well). It is useful in routine repetitive dispensing works.
  • Handling challenging liquids. It is used to dispense viscous, dense, volatile and foaming liquids with better accuracy. Liquids like glycerol, ethanol and whole blood can be handled without much volume error.
  • Assay and PCR setup. It is used for distributing master mix, enzymes and reagents for PCR and ELISA type protocols. It saves time and gives uniform dispensing.
  • Microplate washing. It is used to add wash solution quickly into multiple wells during assay washing steps.
  • Serial dilutions. It is used to make repeated consistent dilutions in experiments. The same step volume can be maintained and error is reduced.
  • Parallel purifications and screens. It is used in doing multiple mini-preps and spin column purification in parallel. It is also used for setting crystallography screening setup.
  • Chromatography and organic synthesis. It is used in preparing samples for HPLC and related works. Airtight tips reduce evaporation and dripping of volatile solvents.
  • Clinical and diagnostic testing. It is used in batch processing works in diagnostics and pharmaceutical testing. It is also applied in environmental and food contaminant testing where many samples are handled together.

Advantages of Repeater Pipettes

  • Increased efficiency and speed. It allows aspiration only once and then multiple equal dispensing is done without refilling again and again. It is very useful in high throughput works like filling multi well plates.
  • Improved ergonomics and reduced strain. It needs less thumb pressing compared to normal pipettes. It reduces fatigue and reduces repetitive strain injury (RSI). Many models have finger hook and in electronic model motor does the plunging work.
  • High accuracy with difficult liquids. It works by positive displacement and no air cushion is present. So viscous gels, volatile solvents, dense liquids and foaming media can be dispensed with better precision.
  • Safety and contamination prevention. The syringe tips have sealed piston and liquid directly contacts piston. Aerosol entry into pipette body is prevented and cross contamination is reduced. It is safer for toxic, infectious and radioactive samples.
  • Reduced plastic and reagent waste. One disposable tip can be used for many dispensing steps (up to around 100 steps). So plastic waste is less compared to changing tips each time. It also reduces dead volume and saves costly reagents.
  • Versatile volume ranges. Different syringe tip sizes can be fitted. A single repeater pipette can cover wide volume range from very small volume (1 µL) up to large volumes (50 mL).

Limitations of Repeater Pipettes

  • Higher consumable costs. The syringe style tips have their own piston inside, so these tips are costly as compared to normal disposable tips.
  • Requires excess liquid (dead volume). Reverse stroke is done and first drop is discarded. A residual stroke also holds some liquid at the end. So more liquid is aspirated than the actual dispensing requirement and it is problem when reagent is less or expensive.
  • Inflexible for variable volumes. It is mainly designed for repeated same volume dispensing. If different volume transfer is needed in every step, it becomes inefficient.
  • Lack of autoclavability. Many repeater pipette instruments and the syringe tips cannot be autoclaved. So sterilization option is limited and mostly chemical cleaning is used.
  • Proprietary tip restrictions. Many models work only with specific brand compatible syringe tips (Combitip, PD-tip). It restricts use of universal cheaper tips and tip recognition may not work with other tips.
  • Operating and chemical restrictions. The pipette and tips have temperature range limits (around 15°C to 40°C). Some chemicals can damage the plastics used in the tip cylinder or piston, so all solvents cannot be used.
  • High tip ejection force. In manual models the syringe tips are bulky and tight. Tip ejection sometimes needs high thumb force and it becomes uncomfortable during repeated use.

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