From:Greg Dobbin <dobbin@Upei.CA>

You sound rather desperate, so maybe my opinion couldn't hurt. If 
your cell suspension is in medium with added protein (like FBS in 
cell culture medium), then I would remove that from the media first, 
(but remember, because I would do it it doesn't necessarily make it 
right! Maybe others have a different idea). This would involve 
washing the cells free of the culture media (ie pellet the cells, 
remove supernate, replace with PBS or plain culture media, and 
repeat). Then resuspend the cells in a similar non-proteinaceous 
media to the desired concentration. 

The next problem can be the salt crystals that form, when the cell 
suspension is left to dry. This is (in my experience) impossible to 
remove completely, but the number crystals and extent of related 
cell damage can be minimized. Place a drop of cell suspension on 
the slide and leave it for 5 mins (less, if drying is noticeable earlier), 
and then use suction (I prefer  to use a glass pasteur pipet hooked 
up to a lab vacuum system) to carefully remove the excess fluid 
(place the pipet at the edge of the drop). Most of the cells would 
have already settled to the glass and "stuck" so that you are 
essentially just removing supernate. Then allow the smears to 
completely dry, and then fix.

 Having said all that, if you have (or have access to) a cytospin 
machine, it makes the best cell smears I've seen. Good luck.

Date sent:      	Wed, 22 Nov 2000 15:37:05 +1100
From:           	Bruce Abaloz <>
Subject:        	HELP - PLEASE!!!!
Forwarded to:

> Hi Dear Histonetters.
> I always couldn't make a good quality smear slide,could you give me some
> idea how to make the smear slide or get a good method to do that from your
> net system.
> What I have done to make the slide is the following: 
> the concentration of the final cell suspension is about 3-6 x 10(6)/ml, 5ul of the cell suspension was dropped on the slide and smeared gently to a small circular area, then air dry and fixed in acetone for 10 min, finally,the fixed slide was stored in -80 degree freezer. 
> But the quality of the slide is not satisfactory for immunocytochemistry.
> Thanks a lot
> ZZ
> Zhen zhang
> Zoology Dept.
> University of Melbourne
> Parkvelle, 3052
> Tel 61-3-8344 4334
> Fax 61-3-8344 7909
> DEPARTMENT of ZOOLOGY	           *  PH:    +61 3 83446282
> The UNIVERSITY of MELBOURNE        *  FAX:   +61 3 83447909 
> Parkville, Victoria.3010           *  EMAIL:                     
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