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Centrifugation

Centrifugation

(Science: procedure) The process of separating fractions of systems in a centrifuge. The most basic separation is to sediment a pellet at the bottom of the tube, leaving a supernatant at a given centrifugal force.

in this case sedimentation is determined by size and density of the particles in the system amongst other factors.

density may be used as a basis for sedimentation in density gradient centrifugation. at very high g values molecules may be separated, i.e. Ultracentrifugation. In continuous centrifugation the supernatant is removed continuously as it is formed.


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centrifugation of hemocytes

I'm not expert in animal biology, but I'd say you do not need EDTA or heparin for resuspension, since the cells are pelleted because of centrifugation, not because they grew there. What about trying some Western or other protein detection after the resuspension to see if there is anything ...

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by JackBean
Thu Jan 30, 2014 9:34 am
 
Forum: Cell Biology
Topic: centrifugation of hemocytes
Replies: 5
Views: 631

Re: centrifugation of hemocytes

I can't see any pellet, BUT today it tried again, after 24h of incubation (cells have time to adhere), and centrifugation at 400 for 5 min, I gently took off the medium and resuspend the cells by pipetting thoroughly up/down: the viability worked well. However, that time i skiped ...

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by Thom
Tue Jan 28, 2014 4:16 pm
 
Forum: Cell Biology
Topic: centrifugation of hemocytes
Replies: 5
Views: 631

Re: centrifugation of hemocytes

Before you wash the cells, can you see any pellet? Wash gently, add your final PBS and pipet to resuspend. Your protocol sounds good to me.

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by jonmoulton
Tue Jan 28, 2014 3:53 pm
 
Forum: Cell Biology
Topic: centrifugation of hemocytes
Replies: 5
Views: 631

centrifugation of hemocytes

... i wash, the cells should stay sticked, unless i resuspend with PBS containing EDTA or heparin... Is it correct? When i take off the medium after centrifugation, and then add PBS are the cell straightforward resuspended or can I take the new added fresh PBS off again until I really resuspend cells ...

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by Thom
Tue Jan 28, 2014 10:50 am
 
Forum: Cell Biology
Topic: centrifugation of hemocytes
Replies: 5
Views: 631

Re: centrifugation of hemocytes

Are you resuspending in a PBS solution isotonic to the cells? Lysis could explain why the cells aren't there. You could try slightly increasing the tonicity of the PBS - better to have crenated cells than lysed cells.

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by jonmoulton
Mon Jan 27, 2014 10:44 pm
 
Forum: Cell Biology
Topic: centrifugation of hemocytes
Replies: 5
Views: 631
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