Re: [Histonet] Safranin O staining

2013-02-19 Thread Victor Wong
Hi Liz,   Get it.  Thank you for the detailed procedures.   Best Regards, Victor From: Elizabeth Chlipala To: Victor Wong ; Tony Henwood (SCHN) ; "histonet@lists.utsouthwestern.edu" Sent: Wednesday, February 20, 2013 12:15 AM Subject: RE: [Histonet] Safranin O staining Victor  

RE: [Histonet] Safranin O staining

2013-02-19 Thread Elizabeth Chlipala
wood (SCHN); histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Safranin O staining Hi Liz, Thank you for your response and your invaluable experience in processing such samples. I'll try manual processing on next batch of samples. I am new to handle these samples. There would be a lot of c

Re: [Histonet] Safranin O staining

2013-02-18 Thread Victor Wong
Sent: Monday, February 18, 2013 11:37 PM Subject: RE: [Histonet] Safranin O staining We have processed and stained these types of samples before.  We do not embed the pellets in agarose prior to processing.  We dye them with eosin and place them in a tea bag and process on a very short cycle - 10 mi

Re: [Histonet] Safranin O staining

2013-02-18 Thread Victor Wong
r sources.   P.S. unforuntuately we don't have a Haematology department but may be we can try other cells.   Best Regards, Victor   From: Tony Henwood (SCHN) To: 'Victor Wong' ; "histonet@lists.utsouthwestern.edu" Sent: Tuesday, February 19, 2013 6:44 AM Subject: RE:

Re: [Histonet] Safranin O staining

2013-02-18 Thread Victor Wong
arose may be deletrious to the staining.  As suggested, I will extend the fixation time.   Best Regards, Victor     From: "koelli...@comcast.net" To: Victor Wong Cc: histonet@lists.utsouthwestern.edu Sent: Monday, February 18, 2013 10:42 PM Subject: Re: [Histonet] Safranin O stain

Re: [Histonet] Safranin O staining

2013-02-18 Thread Victor Wong
ictor Wong ; "histonet@lists.utsouthwestern.edu" Sent: Monday, February 18, 2013 8:42 PM Subject: Re: [Histonet] Safranin O staining Believe it or not, not because you are dealing with cells (small as they are) they require longer than 1 hour to be correctly fixed. This is what I wo

RE: [Histonet] Safranin O staining

2013-02-18 Thread Mitchell Jean A
boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tony Henwood (SCHN) Sent: Monday, February 18, 2013 4:44 PM To: 'Victor Wong'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Safranin O staining Hi Victor, I suppose the take home message from Russ

RE: [Histonet] Safranin O staining

2013-02-18 Thread Tony Henwood (SCHN)
ilto:vhlw...@yahoo.com] Sent: Monday, 18 February 2013 6:36 PM To: Tony Henwood (SCHN); histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Safranin O staining Dear Tony, Thank you for your prompt reply and the paper. I do fix the cell before processing in agarose and after embedding in a

RE: [Histonet] Safranin O staining

2013-02-18 Thread Elizabeth Chlipala
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Victor Wong [vhlw...@yahoo.com] Sent: Monday, February 18, 2013 12:35 AM To: Tony Henwood (SCHN); histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Safranin O staining Dear Tony, Thank you for your prompt reply and the paper. I do fix

Re: [Histonet] Safranin O staining

2013-02-18 Thread koellingr
hen they used.   Ray Koelling Research Scientist University of Washington Seattle - Original Message - From: "Victor Wong" To: histonet@lists.utsouthwestern.edu Sent: Sunday, February 17, 2013 6:34:12 PM Subject: [Histonet] Safranin O staining Hi all, I am workin

Re: [Histonet] Safranin O staining

2013-02-18 Thread Rene J Buesa
"histonet@lists.utsouthwestern.edu" Sent: Sunday, February 17, 2013 9:34 PM Subject: [Histonet] Safranin O staining Hi all, I am working on induced chondrogenesis in cell culture.  After, induction, I put the clump of cells (pellets) in 2% agarose to make a cell block.  When the pellet

Re: [Histonet] Safranin O staining

2013-02-17 Thread Victor Wong
by chondrogenic pellet can be destroyed by heat?  Best Regards, Victor From: Tony Henwood (SCHN) To: 'Victor Wong' ; "histonet@lists.utsouthwestern.edu" Sent: Monday, February 18, 2013 11:45 AM Subject: RE: [Histonet] Safranin O staining Victor, Heat can affect the way ti

Re: [Histonet] Safranin O staining

2013-02-17 Thread Victor Wong
your processing protocol?   Best Regards, Victor   From: Tony Reilly To: "histonet@lists.utsouthwestern.edu" ; Victor Wong Sent: Monday, February 18, 2013 1:50 PM Subject: Re: [Histonet] Safranin O staining Hi Victor I have been using agar cell blocks for over 30 years.  It h

Re: [Histonet] Safranin O staining

2013-02-17 Thread Tony Reilly
Hi Victor I have been using agar cell blocks for over 30 years. It has been my experience that it is better to fix the cells in formalin prior to embedding in the agar to prevent damage to the cells from the heat of the agar. Another important step is to gently heat the agar so that it is ju

[Histonet] Safranin O staining

2013-02-17 Thread Victor Wong
Hi all, I am working on induced chondrogenesis in cell culture.  After, induction, I put the clump of cells (pellets) in 2% agarose to make a cell block.  When the pellet did not sink to the bottom of agarose, I heated to melt the agarose again and centrifuged with higher speed. I fix the agar