[Histonet] Microarray

[ana.merino-trigo]

Hi Lin, 

 

When I began to work with TMA I did have the same problem than you. I got great advices from the designer of the technology, Juha Kononen and also from Stephen Hewitt responsible of the TARP laboratory at the NCI. Basically the best results that I got were following this method: 

 

To build the block: 

 -Use low melting point paraffin for the recipient block 

 -Prior to Construction faced off the recipient block (in this way you will minimize the amount of realignment)




 To cut the block: Sectioning by "TEMPER" the blok

 

1.Once you finish the array place the block - FACE UP- in a oven at 35-37°C overnight 

2.Next AM put it on a cold plate (face up) for an hour 

3.Then reheat at 35-37°C for an hour 

4.Then chill again, in the cold plate for an hour 

5.Repeat the short cycle a few times. 

 

In this way to help to attach properly the spots to the surrounding paraffin, which avoids to lose spots during the microtome cutting. I cut the all block in the same day to avoid loosing slides during the realignment. I used 0.6 mm needles in the automated tissue arrayer ATA-27 from Beecher. By using this method I got a really good yield when using human xenografts and cell lines to build my TMA/CMA blocks. 

 

It's worth to try it, it made a big difference in my results. 

 

Hope this helps, 

Ana

 

 

 

 

 


----- Original Message ----- 
  From: Thom Jensen 
  To: Linresearch <@t> aol.com ; histonet <@t> lists.utsouthwestern.edu 
  Sent: Friday, February 17, 2006 2:47 PM
  Subject: RE: [Histonet] Microarray


  Hey Lin,
  Which arrayer are you using?  If you are using the Chemicon arrayer, their 
  recipient needle is slightly bigger then the donor needle cores.  You will 
  need to set the depth of each recipient hole to match the donor core.  If 
  you are using the Beecher Instrument, then the donor core will fit snuggly 
  in the Recipient hole unless you made the hole larger then the original hole 
  on accident.  Which often happens when people are learning to punch.

  If you are loosing cores from the array block while sectioning the array 
  block on a microtome, then  the problem is you haven't set the punches well. 
    Put the finished array block in an oven on a glass slide (punch surface 
  down on slide) at around 37 to 40 degrees Celsius for 15 minutes then press 
  the glass slide and block together.  This smashes the paraffin around the 
  cores so the cores stick to the paraffin in the array block.  If you don't 
  do this step before you sections the array block the punches will fall out.  
  This step is for both array instruments.

  I hope this helps,

  Cheers,

  Thom Jensen

  For more information of array instruction visit:  www.arrayworkshop.com



  >
  >Hi,
  >Are there any hints to share on getting all the cores to stay evenly in the
  >recipient block and to get sections without losing cores?
  >Lin
  >_______________________________________________
  >Histonet mailing list
  >Histonet <@t> lists.utsouthwestern.edu
  >http://lists.utsouthwestern.edu/mailman/listinfo/histonet



  _______________________________________________
  Histonet mailing list
  Histonet <@t> lists.utsouthwestern.edu
  http://lists.utsouthwestern.edu/mailman/listinfo/histonet



  -- 
  No virus found in this incoming message.
  Checked by AVG Free Edition.
  Version: 7.1.375 / Virus Database: 267.15.11/264 - Release Date: 17/02/2006