Skip to main content
Log in

Quantitative analysis of angiogenesis using confocal laser scanning microscopy

  • Published:
Angiogenesis Aims and scope Submit manuscript

Abstract

Angiogenesis is essential for tumor growth and metastasis. Angiogenesis is commonly quantified by measuring microvessel density (MVD) within tumors. In this report, we compared light microscopy with confocal laser scanning microscopy (CLSM) in the qualitative and quantitative analysis of angiogenesis. MVDs were determined manually in a lung tumor xenograft and a normal skeletal muscle using CD31 immunohistochemical staining and light microscopy. Area of three-dimensional representation of microvessels, detected as CD31 immunofluorescence, was measured automatically using computer-assisted CLSM. By manual counting under light microscopy, the relative level of MVD of the lung tumor vs. skeletal muscle was 0.8. However, the corresponding relative level of microvessels was 3.4 as determined by computer-assisted CLSM. Futhermore, the architecture of microvessels was better delineated with CLSM than with light microscopy. We have applied this CLSM method for analyzing the antiangiogenic effect of an anticancer drug, paclitaxel, in the lung tumor xenograft model. We conclude that CLSM is an appropriate method for quantitative and qualitative analysis of microvasculature in normal and tumor tissues.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Folkman J. Tumor angiogenesis: Therapeutic implications. N Engl J Med 1971; 285: 1182–86.

    Article  PubMed  CAS  Google Scholar 

  2. Weidner N, Semple JP, Welch WR, Folkman J. Tumor angiogenesis and metastasis-correlation in invasive breast carcinoma. N Engl J Med 1991; 324: 1–8.

    Article  PubMed  CAS  Google Scholar 

  3. Belien JAM, Van Diest PJ, Baak JPA. Relationship between vascularization and proliferation in invasive breast cancer. J Pathol 1999; 189: 309–18.

    Article  PubMed  CAS  Google Scholar 

  4. Gimbrone MA, Leapman SB, Cotran RS, Folkman J. Tumor dormancy in vivo by prevention of neovascularization. J Exp Med 1972; 136: 261–76.

    Article  PubMed  Google Scholar 

  5. Vartanian R, Weidner N. Correlation of intratumoral endothelialcell proliferation with microvessel density (tumor angiogenesis) and tumor-cell proliferation in breast carcinoma. Am J Pathol 1994; 144: 1188–94.

    PubMed  CAS  Google Scholar 

  6. Vermeulen PB, Gasparini G, Fox SB et al. Quantification of angiogenesis in solid human tumours: An International Concensus on the methodology and criteria of evaluation. Eur J Cancer 1996; 14: 2474–84.

    Article  Google Scholar 

  7. Jones PH, Harris AL. The current status of clinical trials in antiangiogenesis. Prin Practice Oncol Updates. 2000; 14: 1–6.

    CAS  Google Scholar 

  8. Katoh R, Hemmi A, Komiyama A, Kawaoi A. Confocal laser scanning microscopic observation of angioarchitectures in human thyroid neoplasms. Human Pathol 1999; 30: 1226–31.

    Article  CAS  Google Scholar 

  9. Belien JAM, Somi S, de Jong JS, et al. Fully automated microvessel counting and hot spot selection by image processing of whole tumour sections in invasive breast cancer. J Clin Pathol 1999; 52: 184–92.

    PubMed  CAS  Google Scholar 

  10. ver der Laak JAWM, Westphal JR, Schalkwuk LM, et al. An improved procedure to quantify tumour vascularity using true colour image analysis. Comparison with the manual hot-spot procedure in a human melanoma xenograft model. J Pathol 1998; 184: 136–43.

    Article  Google Scholar 

  11. de Carvalho HF, Taboga SR. Fluorescence and confocal laser scanning microscopy imaging of elastic fibers in hematoxylin-eosin stained sections. Histochem Cell Biol 1996; 106: 587–92.

    Article  PubMed  CAS  Google Scholar 

  12. Lenander C, Holmgren L. A novel method of visualizing vessels in human tumor biopsies. Angiogenesis 1999; 3: 291–3.

    Article  PubMed  CAS  Google Scholar 

  13. Parum DV, Cordell JL, Micklem K, et al. JC70: A new monoclonal antibody that detects vascular endothelium associated antigen on routinely processed tissue sections. J Clin Pathol 1990; 43: 752–7.

    Google Scholar 

  14. Hsu J, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: A comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem Cytochem 1981; 29: 577–80.

    PubMed  CAS  Google Scholar 

  15. Lau DH, Xue L, Young LJ, Burke PA, Cheung AT. Paclitaxel (Taxol): An inhibitor of angiogenesis in a highly vascularized transgenic breast cancer. Cancer Biotherapy Radiopharmaceut 1999; 14: 31–6.

    Article  CAS  Google Scholar 

  16. Lau D, Guo L, Xue L. Anticancer and antiangiogenic efficacy of 7-day versus 21-day schedules of paclitaxel in a lung cancer model. Proc Am Soc Clin Oncol 2001; 20: 107a.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guo, L., Burke, P., Lo, SH. et al. Quantitative analysis of angiogenesis using confocal laser scanning microscopy. Angiogenesis 4, 187–191 (2001). https://doi.org/10.1023/A:1014010801754

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1014010801754

Navigation