Original article
Multispectral Imaging Autofluorescence Microscopy in Colonic and Gastric Cancer Metastatic Lymph Nodes

Preliminary results of this study were accepted for poster presentation in the American Society of Clinical Oncology Gastrointestinal Cancer Symposium in 2004 (Proceedings of the 2004 Gastrointestinal Cancers Symposium ASCO, San Francisco, CA, Jan 2004:59).
https://doi.org/10.1016/j.cgh.2006.11.013Get rights and content

Background & Aims: The lymphadenectomy and extended lymphadenectomy procedures have been points of controversy in surgical oncology. The methods available for the detection of metastatic lymph nodes are numerous. These include lymphoscintigraphy and radiolabeled antibody detection, but in most cancers the currently used technique is sentinel lymph node identification, performed primarily through the use of immunohistochemistry. We propose the application of autofluorescence (AF)-based techniques for lymph node evaluation in colorectal and gastric tumors. Methods: We studied 30 clinical cases: 15 colorectal cancers and 15 gastric cancers. All of the patients were in the advanced stages of the disease and were candidates for adjuvant therapy. Autofluorescence microspectroscopy and multispectral imaging autofluorescence microscopy have been used to analyze the AF emission of metastatic lymph node sections, excited with 365-nm wavelength radiation. The AF spectra were recorded in the range of 400–700 nm. Monochrome AF images were acquired sequentially through interference filters peaked at 450, 550, and 650 nm, and then combined together in a single red-green-blue image. The AF pattern and the emission spectrum of metastatic lymph nodes have unique characteristics that can be used to distinguish them from the normal ones. Results: The results, compared with standard histopathologic procedures and with specific staining methods, supplied a satisfactory validation of the proposed technique, revealing the possibility of improving the actual diagnostic procedures for malignant lymph node alterations. Conclusions: With the development of appropriate instrumentation, the proposed technique could be particularly suitable in intrasurgical diagnosis of metastatic lymph nodes.

Section snippets

Lymph Node Sampling: Recruitment and Treatment of Lymph Nodes

For this study, we enrolled a total of 30 patients: 15 affected by colorectal cancer and 15 affected by gastric cancer. Two control lymph nodes were obtained from informed nonneoplastic patients.

We expressly chose neoplastic patients who were in the advanced stages of the disease and who were candidates for adjuvant therapy. In these patients, the removal of suspected metastatic lymph nodes, independent of location, did not affect the individual patient’s scheduled procedure. None of the

Results

Both multicolored imaging and spectroscopic techniques were used to characterize the AF properties of metastatic lymph nodes and to distinguish them from normal lymph nodes.

Biopsy sections of the lymph nodes, prepared as described in the Materials and Methods section, were processed by AMS and MIAM, sequentially.

Comparison between the AF spectra of metastatic lymph nodes and the control lymph nodes (Figure 1) revealed meaningful differences: the maximum had shifted toward red by more than 20

Discussion

In the past years, the technologic improvements in the fields of excitation sources, light delivery systems, and, in particular, the increase of device sensitivity for the detection and analysis of fluorescence signals, together with a better knowledge of the photo physical characteristics of the endogenous fluorophores, have opened interesting perspectives for applying AF in the characterization of animal tissue features and consequently in diagnosis.

The present study shows that AF analysis

References (63)

  • L. Kretschmer et al.

    Dynamic lymphoscintigraphy and image fusion of SPECT and pelvic CT-scans of aberrant pelvic sentinel lymph nodes in malignant melanoma

    Eur J Cancer

    (2003)
  • M. Monici et al.

    Natural fluorescence of white blood cells: spectroscopic and imaging study

    J Photochem Photobiol B

    (1995)
  • Z. Palasz et al.

    Investigation of normal and malignant laryngeal tissue by autofluorescence imaging technique

    Auris Nasus Larynx

    (2003)
  • R.S. Sohal

    Assay of lipofuscin/ceroid pigment in vivo during aging

    Methods Enzymol

    (1984)
  • Y. Matsumoto

    Lipofuscin pigmentation in pleomorphic adenoma of the palate

    Oral Surg Oral Med Oral Pathol

    (2001)
  • C.E. Canessa et al.

    Anatomic study of lateral pelvic lymph nodes: implications in the treatment of rectal cancer

    Dis Colon Rectum

    (2004)
  • M.L. Quek et al.

    The importance of an extended lymphadenectomy in the management of high grade invasive bladder cancer

    Expert Rev Anticancer Ther

    (2004)
  • P. McCulloch et al.

    Extended versus limited lymph nodes dissection technique for adenocarcinoma of the stomach

    Cochrane Database Syst Rev

    (2003)
  • N. Elias et al.

    Is completion lymphadenectomy after positive sentinel lymph node biopsy for cutaneous melanoma always necessary?

    Arch Surg

    (2004)
  • T. Yokota et al.

    Lymph node metastasis a significant prognostic factor in gastric cancer: a multiple logistic regression analysis

    Scand J Gastroenterol

    (2004)
  • T. Popiela et al.

    Does extended lymphadenectomy, improve survival of pancreatic cancer patients?

    Acta Chir Belg

    (2002)
  • I. Stidili et al.

    Subtotal esophagectomy with extended 2-field lymph node dissection for thoracic esophageal cancer

    Eur J Cardiothorac Surg

    (2003)
  • D. Nitti et al.

    Lumphadenectomy in patients with gastric cancerA critical review

    Suppl Tumori

    (2003)
  • A.E.H. Merrie et al.

    Diagnostic use of the sentinel node in colon cancer

    Dis Colon Rectum

    (2001)
  • E. Pelosi et al.

    Pre-operative lymphatic mapping and intraoperative sentinel lymph node detection in early stage endometrial cancer

    Nucl Med Commun

    (2003)
  • T.J. Polascik et al.

    Comparison of clinical staging algorithms and Indium-capromab pendetide immunoscintigraphy in the prediction of lymph node involvement in high risk prostate carcinoma patients

    Cancer

    (1999)
  • S. Munakata et al.

    Application of immunofluorescence for intraoperative evaluation of sentinel lymph nodes in patients with breast carcinoma

    Cancer

    (2003)
  • G. Babiera et al.

    Lymphatic drainage patterns on early versus delayed breast lymphoscintigraphy performed after injection of filtered Tc-99m sulfur colloid in breast cancer patients undergoing sentinel lymph node biopsy

    Clin Nucl Med

    (2005)
  • J.H. Wong et al.

    Ex vivo sentinel mapping in carcinoma of the colon and rectum

    Ann Surg

    (2001)
  • D.C. Damin et al.

    Sentinel lymph node procedure in patients with epidermoid carcinoma of the anal canal: early experience

    Dis Colon Rectum

    (2003)
  • A. Torne et al.

    The use of sentinel lymph nodes in gynaecological malignancies

    Curr Opin Obstet Gynecol

    (2004)
  • Cited by (10)

    View all citing articles on Scopus

    Supported in part by the Ministero dell’Istruzione della Università e della Ricerca Scientifica grant (2002064155).

    View full text