Hello,
I should transfect THP-1 cells with GFP and use different transfection agents. For this I should quantify the result with DAPI. So, how can I fix THP-1 cells, which are in suspenion, for this?
Thank you for your attention!
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How to fix suspencion cells for DAPI detecting under the microscope?
Started by Sumpf, Apr 27 2009 03:17 AM
6 replies to this topic
#2
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Sunflower
Posted 27 April 2009 - 08:04 AM
if you have access to a cytospin, this is a centrifuge that will allow you to spin suspension cells onto a slide.
"it is a miracle that curiosity survives formal education" -A.E.
#4
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Missele, the little mouse
Posted 28 April 2009 - 12:40 AM
did you try polylysine coated slides? let the cells sediment on the glass (put a drop of 200 µL of cells and let them sediment around 10 min if I remember well), they will attach. then you can wash, label, and fix.
Edited by little mouse, 28 April 2009 - 12:41 AM.
#5
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Posted 28 April 2009 - 02:40 AM
little mouse, on Apr 28 2009, 10:40 AM, said:
did you try polylysine coated slides? let the cells sediment on the glass (put a drop of 200 µL of cells and let them sediment around 10 min if I remember well), they will attach. then you can wash, label, and fix.
I'll try thx
#6
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Missele, the little mouse
Posted 28 April 2009 - 04:48 AM
But I have a suggestion:
if you want to see GFP in THP-1 cells, it would be easier to check this by flow cytometry.
transfect your cells, then fix them with 0.5 % formaldehyde and analyse on a flow cytometer. You will see very fast the percentage of the transfected cells, and the intensity of the fluorescent per each cell.
if you want to see GFP in THP-1 cells, it would be easier to check this by flow cytometry.
transfect your cells, then fix them with 0.5 % formaldehyde and analyse on a flow cytometer. You will see very fast the percentage of the transfected cells, and the intensity of the fluorescent per each cell.
#7
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Posted 28 April 2009 - 05:41 AM
little mouse, on Apr 28 2009, 02:48 PM, said:
But I have a suggestion:
if you want to see GFP in THP-1 cells, it would be easier to check this by flow cytometry.
transfect your cells, then fix them with 0.5 % formaldehyde and analyse on a flow cytometer. You will see very fast the percentage of the transfected cells, and the intensity of the fluorescent per each cell.
if you want to see GFP in THP-1 cells, it would be easier to check this by flow cytometry.
transfect your cells, then fix them with 0.5 % formaldehyde and analyse on a flow cytometer. You will see very fast the percentage of the transfected cells, and the intensity of the fluorescent per each cell.
Even better than slides. Thx
