Flow microfluorometric analysis of nuclear DNA in cells from solid tumors and cell suspensions. A new method for rapid isolation and straining of nuclei.

摘要: A one-step procedure for the preparation of nuclei flow microfluorometric DNA analysis is described. The membranes cells were lysed by non-ionic detergent Nonidet P40. Single-cell suspensions, and specimens solid tissues obtained with fine-needle biopsy, could be prepared equally well as tissue released separately. Lysis was performed in staining solution containing either ethidium bromide or propidium iodide. Fluorescence due to fluorochrome binding RNA, abolished instantaneously presence RNA-se, secondary sites inhibited NaCl. time 10 min samples stable a minimum 12 h. With basic version method, usable, but not always optimal, results all cell types tested: four different mouse ascites tumors, leucocytes, bone-marrow, liver cells, human lymphomas, carcinomas breast lung, mammary carcinoma JB-1 tumor. method further optimized tumour. resulting two modified techniques are Differences leucocytes analogues iodide demonstrated.