Hi all, I am going to do an immunofluorescence microscopy by collecting bacteria-infected cell line at various time points. So I have to collect and fix the samples at different time point and I want to go to view all the samples at once. I would like to know at which step I can stop (after fix, after blocking with BSA or after mounting the cover slip?) and how I can store preserve the samples. The fluorescence dyes I am going to use is FITC and lysotracker
0
2 replies to this topic
#2
LostintheLab
-
- Active Members
-
- 62 posts
Enthusiast
1
Neutral
Neutral
Posted 13 April 2009 - 08:27 PM
Nrelo, on Apr 14 2009, 11:58 AM, said:
Hi all, I am going to do an immunofluorescence microscopy by collecting bacteria-infected cell line at various time points. So I have to collect and fix the samples at different time point and I want to go to view all the samples at once. I would like to know at which step I can stop (after fix, after blocking with BSA or after mounting the cover slip?) and how I can store preserve the samples. The fluorescence dyes I am going to use is FITC and lysotracker
You can either store your cells after you've fixed them, store in PBS or PBS with BSA and sodium azide for a few days in the fridge (sealed with parafilm to prevent drying out). Or after you have completed the staining. I routinely store my slides in the fridge, in a slide box or wrapped in foil to prevent fading.
I'm not sure how far apart your time points are, so you should choose which is the best time schedule for you.
hope that helps
Lost
I knew it! I knew it! Well, not in the sense of having the slightest idea, but I knew there was something I didn't know.
#3
shawnee
-
- Active Members
-
- 5 posts
member
0
Neutral
Neutral
Posted 14 April 2009 - 12:55 AM
Nrelo, on Apr 13 2009, 06:58 PM, said:
Hi all, I am going to do an immunofluorescence microscopy by collecting bacteria-infected cell line at various time points. So I have to collect and fix the samples at different time point and I want to go to view all the samples at once. I would like to know at which step I can stop (after fix, after blocking with BSA or after mounting the cover slip?) and how I can store preserve the samples. The fluorescence dyes I am going to use is FITC and lysotracker
HI, I usually store the cell in 1% bsa in 4oc for up to a week, time concerning immunobloting with primary antibody should optimized empirically range from 1 hour to overnight. I think rodamine glows better than fitc.
