Laboratory centrifuge: Difference between revisions
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Glass centrifuge tubes can be used with most solvents, but tend to be more expensive. They can be cleaned like other [[laboratory glassware]], and can be [[Sterilization (microbiology)|sterilized]] by [[autoclaving]]. Plastic centrifuge tubes, especially microcentrifuge tubes tend to be less expensive. Water is preferred when plastic centrifuge tubes are used. They are more difficult to clean thoroughly, and are usually inexpensive enough to be considered disposable. |
Glass centrifuge tubes can be used with most solvents, but tend to be more expensive. They can be cleaned like other [[laboratory glassware]], and can be [[Sterilization (microbiology)|sterilized]] by [[autoclaving]]. Plastic centrifuge tubes, especially microcentrifuge tubes tend to be less expensive. Water is preferred when plastic centrifuge tubes are used. They are more difficult to clean thoroughly, and are usually inexpensive enough to be considered disposable. |
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Larger samples are spun using centrifuge bottles, which range in capacity from 250 to 750 millilitres. Although some are made of heavy glass, centrifuge bottles are usually made of shatter-proof plastics such as polypropylene or polycarbonate. Sealing closures may be used for added leak-proof assurance. |
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==Safety== |
==Safety== |
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Revision as of 09:02, 3 January 2014
 | This article needs additional citations for verification. (February 2008) |
 A tabletop laboratory centrifuge | |
| Uses | Separation |
|---|---|
| Related items | Gas centrifuge Ultracentrifuge |
A laboratory centrifuge is a piece of laboratory equipment, driven by a motor, which spins liquid samples at high speed. There are various types of centrifuges, depending on the size and the sample capacity.
Like all other centrifuges, laboratory centrifuges work by the sedimentation principle, where the centripetal acceleration is used to separate substances of greater and lesser density.
Types
There are various types of centrifugation:
- Differential centrifugation, often used to separate certain organelles from whole cells for further analysis of specific parts of cells
- Isopycnic centrifugation, often used to isolate nucleic acids such as DNA
- Sucrose gradient centrifugation, often used to purify enveloped viruses and ribosomes, and also to separate cell organelles from crude cellular extracts
There are different types of laboratory centrifuges:
- Microcentrifuges
(devices for small tubes from 0.2 ml to 2.0 ml (micro tubes), up to 96 well-plates, compact design, small footprint; up to 30,000 g)
- Clinical centrifuges
(devices used for clinical applications like blood collection tubes, low-speed devices)
- Multipurpose benchtop centrifuges
(devices for a broad range of tube sizes, high variability, big footprint)
- Stand alone centrifuges
(heavy devices like the ultracentrifuge)
Many centrifuges are available with (refrigerated device) or without cooling function. There are different providers of laboratory centrifuges like Eppendorf, Thermo-Heraeus, Thermo-Sorvall, Hettich, Beckmann-Coulter, MSE, Sigma and AWEL International.
Centrifuge tubes
Centrifuge tubes or centrifuge tips are tapered tubes of various sizes made of glass or plastic. They may vary in capacity from tens of millilitres, to much smaller capacities used in microcentrifuges used extensively in molecular biology laboratories. The most commonly encountered tubes are of about the size and shape of a normal test tube (~ 10 cm long). Microcentrifuges typically accommodate microcentrifuge tubes with capacities from 250 μl to 2.0 ml. These are exclusively made of plastic.
Glass centrifuge tubes can be used with most solvents, but tend to be more expensive. They can be cleaned like other laboratory glassware, and can be sterilized by autoclaving. Plastic centrifuge tubes, especially microcentrifuge tubes tend to be less expensive. Water is preferred when plastic centrifuge tubes are used. They are more difficult to clean thoroughly, and are usually inexpensive enough to be considered disposable.
Larger samples are spun using centrifuge bottles, which range in capacity from 250 to 750 millilitres. Although some are made of heavy glass, centrifuge bottles are usually made of shatter-proof plastics such as polypropylene or polycarbonate. Sealing closures may be used for added leak-proof assurance.
Safety
The load in a laboratory centrifuge must be carefully balanced. This is achieved by using a combination of samples and balance tubes which all have the same weight or by using various balancing patterns without balance tubes.[1] Small differences in mass of the load can result in a large force imbalance when the rotor is at high speed. This force imbalance strains the spindle and may result in damage to the centrifuge or personal injury. Some centrifuges have an automatic rotor imbalance detection feature that immediately discontinues the run when an imbalance is detected.
Before starting a centrifuge, an accurate check of the rotor and lid locking mechanisms is mandatory. Centrifuge rotors should never be touched while moving, because a spinning rotor can cause serious injury. Modern centrifuges generally have features that prevent accidental contact with a moving rotor as the main lid is locked during the run.
Centrifuge rotors have tremendous kinetic energy during high speed rotation. Rotor failure, caused by mechanical stress from the high forces imparted by the motor, can occur due to manufacturing defects, routine wear and tear, or improper use and maintenance. Such a failure can be catastrophic failure, especially with larger centrifuges, and generally results in total destruction of the centrifuge. While centrifuges generally have safety shielding to contain these failures, such shielding may be inadequate, especially in older models, or the entire centrifuge unit may be propelled from its position, resulting in damage to nearby personnel and equipment. Uncontained rotor failures have shattered laboratory windows and destroyed refrigerators and cabinetry. To reduce the risk of rotor failures, centrifuge manufactures specify operating and maintenance procedures to ensure that rotors are regularly inspected and removed from service or derated (only operated at lower speeds) when they are past their expected lifetime.[2]
Another potential hazard is the aerosolization of hazardous samples during centrifugation. To prevent contamination of the laboratory, rotor lids with special aerosol-tight gaskets are available. The rotor can be loaded with the samples within a hood and the rotor lid fixed on the rotor. Afterwards, the aerosol-tight system of rotor and lid is transferred to the centrifuge. The rotor can then be fixed within the centrifuge without opening the lid. After the run, the entire rotor assembly, including the lid, is removed from the centrifuge to the hood for further steps, maintaining the samples within a closed system.
Theory
Protocols for centrifugation typically specify the amount of acceleration to be applied to the sample, rather than specifying a rotational speed such as revolutions per minute. The acceleration is often quoted in multiples of g, the acceleration due to gravity at the Earth's surface. This distinction is important because two rotors with different diameters running at the same rotational speed will subject samples to different accelerations.
The acceleration can be calculated as the product of the radius and the square of the angular velocity.
Relative centrifugal force (RCF) is the measurement of the force applied to a sample within a centrifuge. This can be calculated from the speed (RPM) and the rotational radius (cm) using the following calculation.
- g = RCF = 0.00001118 × r × N2
where:
- g = Relative centrifuge force
- r = rotational radius (centimetre, cm)
- N = rotating speed (revolutions per minute, r/min)
To avoid having to perform a mathematical calculation every time, one can find nomograms for converting RCF to rpm for a rotor of a given radius. A ruler or other straight edge lined up with the radius on one scale, and the desired RCF on another scale, will point at the correct rpm on the third scale. Example Based on automatic rotor recognition, up to date centrifuges have a button for automatic conversion from RCF to rpm and vice versa.
See also
References
- ^ Frothingham, R (February 1999). "Centrifugation without a balance tube". American Laboratory. 31 (19). Shelton, CT: International Scientific Communications: 10. ISSN 0044-7749. Retrieved March 18, 2011.
{{cite journal}}: Cite has empty unknown parameter:|coauthors=(help) - ^ Cornell University EH&S. "Centrifuge Accident". Retrieved 29 December 2010.
External links
