From: Manabu Takahashi (manabu32@ymg.urban.or.jp)
Date: Sun Dec 15 1996 - 10:02:33 EST
Dr. Ormerod wrote : >The cells were then washed and fixed in 70% ethanol on ice. After 1 to 24 >hours, the cells were washed and resuspendedin a buffer containing PI and >RNase and incubated at 37 for 30 min. >We consistently obtain two peaks in the DNA histogram (30% difference). >If we use a detergent method to prepare unfixed nuclei, there is only one peak >in the DNA histogram. > Dr. M. Ormerod We noticed this several years ago when we examined our blood in our department. Some showed two peaks but some others do not. All of the examinee were hematologically normal. In our experience, the DIs were less than 20% apart. We used a detergent method to prepare unfixed nuclei. Another experience: When leukocytes are fixed in 70% ethanol that contains RNase, some of the cells move out of the G1 peak to form several peaks of lower DIs. The process is slow and requires a month or two before peaks are formed. An important requirement is to use a very dilute PI solution for staining. Manabu ****************************** Dr. Manabu Takahashi Professor emeritus Yamaguchi University, Ube, Japan e-mail: manabu32@ymg.urban.or.jp ******************************
This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:31:33 EST
