Isolated cell disruption methods?

Andrew Dueck ADDUECK at BIOLOGY.watstar.uwaterloo.ca
Thu Aug 11 13:45:12 EST 1994


In article <mbxfd-100894150958 at macfd.biochem.nottingham.ac.uk> mbxfd at unicorn.nott.ac.uk (Fergus Doherty) writes:
>From: mbxfd at unicorn.nott.ac.uk (Fergus Doherty)
>Subject: Isolated cell disruption methods?
>Date: 10 Aug 1994 14:10:17 GMT

>Hello,
>Can anyone give advice on the relative merits of:

>* the nitrogen cavitation bomb
>* the 'cell disrupter' from the EMBL workshop (a steel block wih a narrow 
>channel with a ball bearing in it around which a cell suspensio is forced
>with a syringe)
>* the 'mini bead-beater' (a device for violently agitating a cell
>suspension with glass beads)
>*  any other method

>for disrupting isolated cells (ie culture cells) to obtain homogenates
>suitable for isolation of intact organelles (ie lysosomes)?

>If you know of the EMBL cell disrupter can you tell me who to contact there
>(e-mail if possible) or if there is another source for this device.  Any
>other recommendations appreciated, bearing in mind the cells in questions
>are small (monocytes, RBL cells and reticulocytes) and have proven
>difficult/impossible to homogenize by other methods (eg Dounce).

>Fergus Doherty,
>dept. Biochemistry,
>University Medical School,
>Queen's Medical Centre,
>Nottingham NG7 2UH
>Tel: 602 709366  FAX 602 422225 Internet: mbxfd at unicorn.nott.ac.uk

	If you are trying to disrupt the cell membrane, I would suggest the 
use of 0.01% Sodium Dodecyl Sulfate.  I'm not sure what this would do to 
organelles with respect to whether or not they would remain intact.
	Another method to try might be the use of a sonicator.  Again, you 
may have to fiddle with the settings so that it does not destroy the 
lysosomes.
	To isolate lysosomes, you may want to try the technique of 
differential centrifugation.  I think it should work effectively and easily 
for you.
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Andrew Dueck






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