USA Scientific目录号#1412-1410的管子自发荧光问题

niwanmao

NIH疫苗研究中心的Mario Roederer博士昨天一封email中提到，USA Scientific一批目录号#1412-1410的管子（见http://www.usascientific.com/1.2 ... racked-sterile.aspx），经过γ射线照射之后，做流式标记出现明显的自发荧光，主要发生在V450通道（近紫外激光405nm激发，450nm发射），该通道主要检测Pacific Blue、ViViD、Brilliant Violet 450等染料。问题管主要集中在目录号为 #1412-1410，而目录号为#1412-0400的没问题。

问题管

不知道站内有没有朋友在用这批管子，在此翻译并转发提示一下！

原信如下：

Dear all --

we recently discovered that one type of "bullet" tube we use for staining leads to large increases in autofluorescence, primarily in the "V450" channel (i.e., excited by near-UV laser, 405 nm, and emitting in the 450 nm range).  This is the detector you would use for Pacific Blue, ViViD, or Brilliant Violet 450 dyes.

Interestingly, the problem only occurs in the tubes that have been sterilized by gamma-irradiation -- the unsterilized tubes are fine.  The tubes are polypropylene, so it's quite possible that this issue may extend to other tubes that are similarly sterilized.

The specific tubes come from USA Scientific.  The "problem tubes" are catalog #1412-1410; the acceptable tubes are catalog #1412-0400.

The sterilization process must be leading to the generation of a UV-excitable dye which leaches into the cells.  The longer you incubate cells in the "problem" tubes, the brighter they become.  Curiously, this doesn't always happen uniformly -- sometimes half the cells become bright and half do not.  Perhaps it requires close proximity of the cell to the plastic.  This heterogeneity is insidious -- it looks like you are generating new subsets of cells (rather than having a staining artefact).

mr

_______________________________________________
Mario Roederer, Ph.D.
Senior Investigator, ImmunoTechnology Section
Vaccine Research Center, NIAID, NIH
40 Convent Dr., Room 5509
Bethesda, MD 20892-3015
Phone: 301 594-8491
FAX: 301 480-2788