Micropipette and Cuvet- standard operating procedure

Contains standard operating procedures for micropipettes, their use, and their working principles. Also contains details about cuvet, their types, and their uses.

1. Micropipette and Cuvet Standard Operating Procedure Presenter - Dr. Pranav U S (JR3) Peer support - Dr. Shreedhar Angadi (JR3) Department of Pharmacology & Therapeutics King George’s Medical University, Lucknow, U.P., India E-mail: [email protected]

2. Micropipette Micropipettes or pipettes, are instruments used to measure liquid ranging between volumes of  1-10000 µl  and transfer it from one sample container to another. This is a basic liquid-handling instrument for almost all scientific laboratories.

3. Number of Channels Single Channel Pipette Multi Channel Pipette

4. Operating Mechanism Mechanical Pipette Electronic Pipette

5. Components Disposable Tips

6. Components Pipette body: The main housing of the micropipette which houses the internal mechanisms Plunger: This is the mechanism that allows you to pull up and dispense liquids. Pressing down the plunger will expel air via the tip. Releasing this plunger will then pull up solution into the tip. Tip ejector: Pushing this button will remove of disposable tip, ensuring contamination-free transfers.

7. Volume adjustment ring: This allows precise selection of the desired volume to be aspirated and dispensed. The volume that will be pulled into the tip is shown on the volume display. Volume display: A digital or analog display showing the dispense volume. Tip Cone: You can position a disposable tip onto the tip cone. If you are using the correct tip, a disposable tip should fit on simply by pressing the tip onto the micropipette.

8. Model Volume Range P10 P20 P100 P200 P1000 P5000 P10000 1–10 µl 2–20 µl 10–100 µl 20–200 µl 100–1000 µl 500–5000 µl 1000–10000 µl

9. Operating Procedure Insert the pipette tip into the tip cone making sure it is securely in place Holding the pipette vertically, push down the control button until the first stop Immerse the pipette tip into your solution ( I mmersed depth should be between 2-5 mm, based upon the volume of the tip) Slowly release the control button until it is at its resting position

10. Wait 2-3 seconds, then withdraw the tip from the solution Place the tip inside the vial you wish to dispense into, press the control button to the first stop, and then finally fully press down the button to dispense the remaining solution Withdraw the pipette from the vial and release the control button After finishing with your pipette firmly press the ejector button to remove the tip

11. Volume Adjustment To adjust the volume of the micropipette, rotate the volume adjustment knob. To increase the volume, turn the knob anti-clockwise to. To decrease the volume, turn the knob clockwise. Calibration Each micropipette is pre-calibrated and includes a manufacturer’s inspection certificate to verify the accuracy and repeatability values Over time, the micropipette may drift from these values due to excessive use or mishandling of the micropipette.

12. Applications Molecular Biology: Used for precise measurement and transfer of DNA, RNA, and proteins during experiments like PCR (Polymerase Chain Reaction) and gel electrophoresis. Microbiology: Handling small volumes of bacterial cultures, media, and reagents for various assays and experiments. Pharmaceutical Research: Accurate dosing and mixing of chemical compounds, drugs, and solutions during drug development and testing.

13. Clinical Laboratories: Measuring and transferring small volumes of blood, serum, and other clinical samples for diagnostic tests Forensic Analysis: Handling trace amounts of biological samples, chemicals, and reagents during forensic investigations Environmental Testing: Measuring pollutants, chemicals, and other environmental samples in small quantities. Biotechnology: Transferring small volumes of enzymes, substrates, and other reagents during biotechnological processes and research.

14. Cuvet (or cuvette) It is a small, typically rectangular or cylindrical, optical container used in laboratories for holding liquids that are to be analyzed using spectrophotometry or similar techniques Material : Typically made of glass , quartz , or plastic depending on the type of analysis and wavelength of light being used Common cuvette volumes range from 70 microliters to 3.5 milliliters

15. Function of a Cuvet Used to hold liquid samples for optical measurements , typically in UV-visible spectrophotometers . The liquid sample in the cuvet is exposed to a beam of light, and the absorbance or transmittance of the light is measured to determine properties like concentration of solutes. Standard 3.5mL Cuvettes

16. Semi Micro Cuvettes Sub Micro Cuvettes

17. Types of Cuvets Type Wavelength Range Common Use Glass Cuvets 400-700 nm (Visible) Visible light spectrophotometry Quartz Cuvets 200-2500 nm (UV to IR) UV-visible spectroscopy Plastic Cuvets 400-700 nm (Visible) Routine, economical measurements Optical Cuvets 340-2500 nm Precision analyses, fluorescence

18. Cuvet Size and Volume Standard Path Length : 10 mm (optimal for most measurements) Custom Sizes : 5 mm (high conc.) or 20 mm (low conc.) Path Length Use Example 10 mm Standard spectrophotometry General analysis 5 mm High concentration samples Concentrated samples 20 mm Low concentration samples Environmental testing

19. Applications of Cuvets Spectrophotometry : Measuring concentration of solutes (e.g., proteins, nucleic acids) Clinical Chemistry : Analysing analytes in blood/urine Pharmaceutical : Drug concentration/purity analysis Environmental Testing : Measuring pollutants in air/water

20. References Ossila . How to Use a Micropipette [Internet]. Available from: https://www.ossila.com/pages/how-to-use-micropipette Accumax . A Complete Guide on Micropipettes [Internet]. Available from: https://www.accumaxlab.com/micropipette-guide/ Mxrady . Understanding Cuvettes: The Essential Tool in Spectroscopy. MxRady.Available from: https://mxrady.com/understanding-cuvettes-the-essential-tool-in-spectroscopy/

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