Dispersal of solid tumors for flow cytometer (FCM) analysis
Flow cytometer (FCM) analysis of the DNA content of solid tumors necessitates tumor dispersal into single cell suspensions. Single cell suspensions of S102F mammary tumors are prepared by incubating minced tumor tissue at 37/sup 0/C in a sucrose-salt solution containing sodium tetraphenylboron (TPB), a potassium-complexing reagent. After one hour of incubation, the tumor pieces are passed several times through a plastic syringe to dissociate the remaining undispersed fragments. The cells are incubated for an additional 30 minutes and then filtered through 37 ..mu..m nylon mesh. The tumor cells are washed with saline and fixed in formalin for FCM analysis. Approximately 2 to 8 x 10/sup 7/ cells are obtained per 150 ..mu..l tumor volume. Histograms of the distribution of the DNA content of acriflavin-stained tumor cells show a minimal amount of background debris and cell clumping.
- Research Organization:
- California Univ., Livermore (USA). Lawrence Livermore Lab.
- DOE Contract Number:
- W-7405-ENG-48
- OSTI ID:
- 6790066
- Report Number(s):
- UCRL-80105; CONF-770324-9
- Resource Relation:
- Conference: 3. international conference on impulse cytophotometry, Vienna, Austria, 30 Mar 1977
- Country of Publication:
- United States
- Language:
- English
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