Lidstrom:Sorvall Centrifuge: Difference between revisions
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== | == Background == | ||
[[image:Lidstrom_Sorvoll.png|thumb|upright=1.0|center|Lidstrom Sorvoll centrifuge]] | [[image:Lidstrom_Sorvoll.png|thumb|upright=1.0|center|Lidstrom Sorvoll centrifuge]] | ||
We have a Sorvoll RC-5B centrifuge that is useful for spinning large volumes in 0.5L bottles or medium-sized (a few mL) volumes very quickly | |||
== Rotors == | == Rotors == | ||
[http://www.geminibv.nl/labware/sorvall-gsa-rotor/sorvall-rotor-gsa-gs3-sa600-se12-sm24-ss34-manual.pdf Sorvall rotors] | |||
[[image: | |||
=== SS-34 === | |||
* Holds 8*50 mL, spins at '''[http://www.geminibv.nl/labware/sorvall-gsa-rotor/sorvall-rotor-gsa-gs3-sa600-se12-sm24-ss34-manual.pdf 20,000 RPM/36,900 RCF]''' | |||
** BUT Katya (circa 2010) recommends using it at <= 18,000 RPM | |||
**Tubes: [[image:Lidstrom_SS-34 tubes.png|thumb|upright=0.3|center|SS-34 tubes]] | |||
***Can't be autoclaved. | |||
**** Actually, Amanda might have autoclaved them before. | |||
***Can't write on body; can only write on cap. Sandy says stickers don't stick. | |||
[[image:SS-34 rotor speeds and k factors.png|thumb|upright=2.0|center|SS-34 rotor speeds and k factors]] | [[image:SS-34 rotor speeds and k factors.png|thumb|upright=2.0|center|SS-34 rotor speeds and k factors]] | ||
=== GS-3 === | |||
* [http://www.thermoscientific.fr/eThermo/CMA/PDFs/Various/File_582.pdf GS-3]. | |||
** BUT Katya (circa 2010) recommends using it at <= 7,500 | |||
** For bigger bottles. (0.5L) | |||
** Holds eppendorf (1.5mL) tubes | |||
* Spins at '''[http://www.geminibv.nl/labware/sorvall-gsa-rotor/sorvall-rotor-gsa-gs3-sa600-se12-sm24-ss34-manual.pdf 9,000 RPM/13,700 RCF]''' | |||
* Requires 3 pieces for installation: | |||
*** (1) the base that you put the filled bottles in | |||
*** (2) a matching black metal lid | |||
*** (3) a metal rod with threads at one end and a knob on the other. | |||
** Place the based down, screw the lid on '''counter clockwise''' and then use the pin to secure it. Try lifting to ensure you secured it correctly. | |||
[[image:GS-3 rotor speeds and k factors.png|thumb|upright=2.0|center|GS-34 rotor speeds and k factors]] | [[image:GS-3 rotor speeds and k factors.png|thumb|upright=2.0|center|GS-34 rotor speeds and k factors]] | ||
== Explanation of RCF == | |||
[[image:RCF_explanation.png|thumb|upright=2.0|center|RCF explanation]] | [[image:RCF_explanation.png|thumb|upright=2.0|center|RCF explanation]] | ||
* For reference, the benchtop 50mL centrifuges (e.g. [https://static.thermoscientific.com/images/D21714~.pdf ST-15R] with a [https://extranet.fisher.co.uk/webfiles/fr/Pjointes/Documentation/SOR007_EN%20SORVALL%20ST16&ST40%20CENTRIFUGE.pdf TX-400] swinging bucket rotor) we have spin at 4696 RCF. | |||
*There are stickers with maximum speeds. These were set by Katya, a Lidstrom Lab manager, circa 2011. It is unknown what the motivation was for keeping the rotors below maximum speed. | |||
[[image:Lidstrom_sorvoll_speed_instructions.png|thumb|upright=2.0|center|Lidstrom sorvoll speed instructions]] | |||
== Instructions == | |||
# pre-chill the rotor you plan to use (overnight in fridge) | |||
# turn on centrifuge and set it to the desired temperature range | |||
## The dial on the left (under the temperature gauge) is really two knobs; one turns the lower bound blue stick and the other turns the upper bound red stick. | |||
## The centrifuge will start cooling as soon as you turn it on. | |||
# Install the rotor of your choice | |||
## The small SS-34 rotor requires a T-shaped piece of metal with a screw end on it to install. Turn '''counter clockwise''' to screw it on. Use the same piece to unscrew it upon completion. | |||
## The large GS-3 rotor has 3 pieces: the base that you put the filled bottles in, a lid, and a metal rod with threads at one end and a knob on the other. Place the based down, screw the lid on '''counter clockwise''' and then use the pin to secure it. Try lifting to ensure you secured it correctly. | |||
# Fill bottles with the cells you want to centrifuge: | |||
## Bottles for the GS-3 are in the prepared media cabinet at the bottom (1/2015 [[Users:Janet B. Matsen|JM]]) | |||
## Bottles for the SS-34 are on the dishwasher room shelves (1/2015 [[Users:Janet B. Matsen|JM]]) | |||
# To turn on, you have to have the black line in the temperature gauge panel between the blue and red lines. Then the start button glows and you can begin spinning. | |||
## Note: the centrifuge is slow to speed up and slow to lose speed as well. | |||
# Remove centrifuged cultures | |||
## The machine slows down slowly. | |||
## Leave the lid of the centrifuge open for a few hours until it warms up to room temperature and any condensation has evaporated. This will reduce odors. | |||
== Sample protocols == | |||
=== Example protocol for SIP3-4: === | |||
Janet's SIP3-4 centrifugation: | |||
* Chill to 4<sup>o</sup>C with SS-34 rotor inside | |||
* Fill correct tubes | |||
* Spin for ___ min at ___ RCF at ___<sup>o</sup>C | |||
=== Example protocol for AM1: === | |||
Ask Nate/Ceci | |||
== Centrifugation Math & Basics == | |||
* [http://www.sigmaaldrich.com/technical-documents/articles/biofiles/centrifugation-basics.html Sigma summary]: | |||
** sedimentation rate is proportional to RFC, and proportional to the square of the particle diameter (cell diameter). | |||
** [[image:Lidstrom_Stokes_equation.png|thumb|upright=2.0|center|Stoke's equation]] | |||
** [[image:Lidstrom centrifugation speeds for applications.png|thumb|upright=2.0|center|centrifugation speeds for applications]] |
Revision as of 11:27, 26 January 2015
Back to Protocols
Background
We have a Sorvoll RC-5B centrifuge that is useful for spinning large volumes in 0.5L bottles or medium-sized (a few mL) volumes very quickly
Rotors
SS-34
- Holds 8*50 mL, spins at 20,000 RPM/36,900 RCF
- BUT Katya (circa 2010) recommends using it at <= 18,000 RPM
- Tubes:
- Can't be autoclaved.
- Actually, Amanda might have autoclaved them before.
- Can't write on body; can only write on cap. Sandy says stickers don't stick.
- Can't be autoclaved.
GS-3
- GS-3.
- BUT Katya (circa 2010) recommends using it at <= 7,500
- For bigger bottles. (0.5L)
- Holds eppendorf (1.5mL) tubes
- Spins at 9,000 RPM/13,700 RCF
- Requires 3 pieces for installation:
- (1) the base that you put the filled bottles in
- (2) a matching black metal lid
- (3) a metal rod with threads at one end and a knob on the other.
- Place the based down, screw the lid on counter clockwise and then use the pin to secure it. Try lifting to ensure you secured it correctly.
Explanation of RCF
- For reference, the benchtop 50mL centrifuges (e.g. ST-15R with a TX-400 swinging bucket rotor) we have spin at 4696 RCF.
- There are stickers with maximum speeds. These were set by Katya, a Lidstrom Lab manager, circa 2011. It is unknown what the motivation was for keeping the rotors below maximum speed.
Instructions
- pre-chill the rotor you plan to use (overnight in fridge)
- turn on centrifuge and set it to the desired temperature range
- The dial on the left (under the temperature gauge) is really two knobs; one turns the lower bound blue stick and the other turns the upper bound red stick.
- The centrifuge will start cooling as soon as you turn it on.
- Install the rotor of your choice
- The small SS-34 rotor requires a T-shaped piece of metal with a screw end on it to install. Turn counter clockwise to screw it on. Use the same piece to unscrew it upon completion.
- The large GS-3 rotor has 3 pieces: the base that you put the filled bottles in, a lid, and a metal rod with threads at one end and a knob on the other. Place the based down, screw the lid on counter clockwise and then use the pin to secure it. Try lifting to ensure you secured it correctly.
- Fill bottles with the cells you want to centrifuge:
- To turn on, you have to have the black line in the temperature gauge panel between the blue and red lines. Then the start button glows and you can begin spinning.
- Note: the centrifuge is slow to speed up and slow to lose speed as well.
- Remove centrifuged cultures
- The machine slows down slowly.
- Leave the lid of the centrifuge open for a few hours until it warms up to room temperature and any condensation has evaporated. This will reduce odors.
Sample protocols
Example protocol for SIP3-4:
Janet's SIP3-4 centrifugation:
- Chill to 4oC with SS-34 rotor inside
- Fill correct tubes
- Spin for ___ min at ___ RCF at ___oC
Example protocol for AM1:
Ask Nate/Ceci
Centrifugation Math & Basics
- Sigma summary:
- sedimentation rate is proportional to RFC, and proportional to the square of the particle diameter (cell diameter).