Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Cell Biology and Cell Culture > Flow Cytometry Forum
Register Search Today's Posts Mark Forums Read

Flow Cytometry Forum Flow Cytometry Forum. Discuss FACS cell sorting, flow cytometry, analysis and cell staining.


Prevent Neutrophil Cells from Iatrogenic Activation

Prevent Neutrophil Cells from Iatrogenic Activation - Flow Cytometry Forum

Prevent Neutrophil Cells from Iatrogenic Activation - Flow Cytometry Forum. Discuss FACS cell sorting, flow cytometry, analysis and cell staining.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 01-28-2009, 04:25 AM
Pipette Filler
Points: 2,176, Level: 29 Points: 2,176, Level: 29 Points: 2,176, Level: 29
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2006
Posts: 7
Thanks: 0
Thanked 0 Times in 0 Posts
Exclamation Prevent Neutrophil Cells from Iatrogenic Activation



Prevent Neutrophil Cells from Iatrogenic Activation:
Hi, Im currently analyzing neutrophils with flouroscences-conjugated anti-CD18/41/61 antibodies by BD Flow cytometer. However, the neutrophils from control mice are always activated!

I use 3.8% sodium citrate as anti-coagulate and 0.5%(v/v) paraformaldehyde for fixation.

I received a lot of positive anti-CD18/41/61 signals in control mice, especially anti-CD18 and my boss was mad and I was very sad .

If anyone knows how to prevent iatrogenic activation of neutrophils, please please help me. Anything, any little tricks might help. Thank you very much.
Reply With Quote
  #2  
Old 01-28-2009, 04:26 AM
Pipette Filler
Points: 2,407, Level: 31 Points: 2,407, Level: 31 Points: 2,407, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2006
Posts: 17
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Prevent Neutrophil Cells from Iatrogenic Activation

3 suggestions for you:

1) Use 3.2%Citrate (dilute 1:9(blood)), look the attached article.

2) Use citrated blood directly for fax analysis (if you are using isolated neutrophils), so that u can be sure your not messing up while doing ur experiment (n to avoid unnecessary steps). if this process does not work then u need to go for fixing with parafarmaldehyde.

3) use RT and on ice conditions for your experiments (to catch the expected results somewhere rather repeating the experiment with different variables)
Reply With Quote
  #3  
Old 01-28-2009, 04:27 AM
Pipette Filler
Points: 2,448, Level: 31 Points: 2,448, Level: 31 Points: 2,448, Level: 31
Activity: 33% Activity: 33% Activity: 33%
 
Join Date: Jan 2006
Posts: 27
Thanks: 0
Thanked 5 Times in 5 Posts
Default Re: Prevent Neutrophil Cells from Iatrogenic Activation

i was using 1% parafarmaldehyde for monocyte surface receptor detection, i used to freeze 1% PFA at -20C and used for 4-5 times in 2 weeks.
i had expected results.

have you try using direct blood for cd18 detection?
Reply With Quote
Reply

Tags
activation , cells , iatrogenic , neutrophil , prevent


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
mouse hepatocytes isolation amicia Cell Biology and Cell Culture 39 05-11-2012 11:52 AM
Characteristics of cancerous cells aftabac Cell Biology and Cell Culture 2 08-26-2011 11:40 PM
Human Cytome Project - Update 24 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 1 08-01-2010 02:18 PM
How to isolate adult from circulation? peterish Stem Cell Forum 0 05-19-2008 10:02 PM
Human Cytome Project - Update 6 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 0 01-06-2005 11:18 AM


All times are GMT. The time now is 04:11 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.16043 seconds with 16 queries