FLOW CYTOMETRIC ANALYSIS OF DNA AND NUCLEAR-PROTEIN IN PARAFFIN-EMBEDDED TISSUE

被引:9
作者
CIANCIO, G [1 ]
POLLACK, A [1 ]
BLOCK, NL [1 ]
机构
[1] UT,MD ANDERSON CANC CTR,DEPT RADIOTHERAPY,HOUSTON,TX
来源
CYTOMETRY | 1993年 / 14卷 / 02期
关键词
DNA; NUCLEAR PROTEIN; PARAFIN-EMBEDDED; FLOW CYTOMETRY;
D O I
10.1002/cyto.990140213
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Previously we described the simultaneous quantification of DNA and nuclear protein in unfixed tissue from solid tumors. The resultant 2 parameter flow cytometric analysis has several advantages over that of DNA alone. In this report, we describe a modification of the technique for the analysis of formalin-fixed paraffin-embedded tissue. Paraffin-embedded material was prepared by hydrating sections, incubating in 0.5% pepsin solution, washing, and resuspending in buffer containing nonionic detergent. The nuclei were then stained with fluorescein isothiocyanate and propidium iodide in the presence of ribonuclease. Several solid tumor tissue types have been analyzed, including breast, colon, kidney, and thymus. The best results were obtained when the initial pepsin treatment was for 1.5 h, instead of 0.5 h. Pepsin treatment for 1.5 h improved the CVs of both the DNA and nuclear protein parameters, and did not appear to reduce nuclear protein levels or to cause significant disintegration of nuclei. The DNA/nuclear protein histograms of unfixed and fixed, paraffin-embedded tissue were similar. Since tumor nuclei typically have higher protein levels than DNA-diploid nuclei, the technique reduces population over-lapping and permits less subjective identification of DNA aneuploidy.
引用
收藏
页码:205 / 209
页数:5
相关论文
共 8 条
  • [1] FLOW-CYTOMETRY USING PARAFFIN-EMBEDDED TISSUE - 5 YEARS ON
    HEDLEY, DW
    [J]. CYTOMETRY, 1989, 10 (03): : 229 - 241
  • [2] METHOD FOR ANALYSIS OF CELLULAR DNA CONTENT OF PARAFFIN-EMBEDDED PATHOLOGICAL MATERIAL USING FLOW-CYTOMETRY
    HEDLEY, DW
    FRIEDLANDER, ML
    TAYLOR, IW
    RUGG, CA
    MUSGROVE, EA
    [J]. JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1983, 31 (11) : 1333 - 1335
  • [3] EFFECTS OF SEVERAL COMMONLY USED FIXATIVES ON DNA AND TOTAL NUCLEAR-PROTEIN ANALYSIS BY FLOW-CYTOMETRY
    HERBERT, DJ
    NISHIYAMA, RH
    BAGWELL, CB
    MUNSON, ME
    HITCHCOX, SA
    LOVETT, EJ
    [J]. AMERICAN JOURNAL OF CLINICAL PATHOLOGY, 1989, 91 (05) : 535 - 541
  • [4] DIFFERENT OPINIONS ON CLASSIFICATION OF DNA HISTOGRAMS PRODUCED FROM PARAFFIN-EMBEDDED TISSUE
    JOENSUU, H
    KALLIONIEMI, OP
    [J]. CYTOMETRY, 1989, 10 (06): : 711 - 717
  • [5] FLOW-CYTOMETRY, CELLULAR DNA CONTENT, AND PROGNOSIS IN HUMAN MALIGNANCY
    MERKEL, DE
    DRESSLER, LG
    MCGUIRE, WL
    [J]. JOURNAL OF CLINICAL ONCOLOGY, 1987, 5 (10) : 1690 - 1703
  • [6] QUANTITATION OF CELL KINETIC RESPONSES USING SIMULTANEOUS FLOW CYTOMETRIC MEASUREMENTS OF DNA AND NUCLEAR-PROTEIN
    POLLACK, A
    MOULIS, H
    BLOCK, NL
    IRVIN, GL
    [J]. CYTOMETRY, 1984, 5 (05): : 473 - 481
  • [7] Pollack A, 1990, Methods Cell Biol, V33, P315
  • [8] MEASUREMENT VARIABILITY IN DNA FLOW-CYTOMETRY OF REPLICATE SAMPLES
    WHEELESS, LL
    COON, JS
    COX, C
    DEITCH, AD
    WHITE, RWD
    KOSS, LG
    MELAMED, MR
    OCONNELL, MJ
    REEDER, JE
    WEINSTEIN, RS
    WERSTO, RP
    [J]. CYTOMETRY, 1989, 10 (06): : 731 - 738