Multispectral imaging: a review of its technical aspects and applications in anatomic pathology.

The field of anatomic pathology has changed significantly over the last decades and, as a result of the technological developments in molecular pathology and genetics, has had increasing pressures put on it to become quantitative and to provide more information about protein expression on a cellular level in tissue sections. Multispectral imaging (MSI) has a long history as an advanced imaging modality and has been used for over a decade now in pathology to improve quantitative accuracy, enable the analysis of multicolor immunohistochemistry, and drastically reduce the impact of contrast-robbing tissue autofluorescence common in formalin-fixed, paraffin-embedded tissues. When combined with advanced software for the automated segmentation of different tissue morphologies (eg, tumor vs stroma) and cellular and subcellular segmentation, MSI can enable the per-cell quantitation of many markers simultaneously. This article covers the role that MSI has played in anatomic pathology in the analysis of formalin-fixed, paraffin-embedded tissue sections, discusses the technological aspects of why MSI has been adopted, and provides a review of the literature of the application of MSI in anatomic pathology.

Infrared spectroscopy: shedding light on synovitis in patients with rheumatoid arthritis.

OBJECTIVES: It is difficult to determine the extent of synovial involvement early in the course of rheumatoid arthritis. A spectroscopic technique was used to characterize the synovium of the small finger joints in both early and late rheumatoid arthritis. This synovium was also compared against normal joints. METHODS: Near-infrared spectroscopy assesses the absorption of near-infrared light by specific joints, giving a characteristic "fingerprint" of the properties of the underlying tissues. Triple measurements by infrared spectroscopy were taken at the bilateral second and third metacarpophalangeal joints. Multivariate analysis was applied. RESULTS: Analysis was able to demonstrate relationships between the specific sources of spectral variation and joint tenderness or swelling as well as radiographic damage. Further use of multivariate analysis allowed recognition of the spectral patterns seen in early disease vs late rheumatoid arthritis and correct classification of over 74% of the joints. CONCLUSIONS: The spectral regions where differences occurred were in the absorption bands related to tissue oxygenation status, allowing the provocative implication that this technique could be detecting ischaemic changes within the joint. Near-infrared spectroscopy may thus be able to provide us with some information about the biochemical changes associated with synovitis.

Towards non-invasive screening of skin lesions by near-infrared spectroscopy.

A noninvasive tool for skin tumor diagnosis would be a useful clinical adjunct. The purpose of this study was to determine whether near-infrared spectroscopy can be used to noninvasively characterize skin lesions. In vivo visible- and near-infrared spectra (400--2500 nm) of skin neoplasms (actinic keratoses, basal cell carcinomas, banal common acquired melanocytic nevi, dysplastic melanocytic nevi, actinic lentigines, and seborrheic keratoses) were collected by placing a fiberoptic probe on the skin. Paired t tests, repeated measures analysis of variance and linear discriminant analysis were used to determine whether significant spectral differences existed and whether spectra could be classified according to lesion type. Paired t tests showed significant differences (p < 0.05) between normal skin and skin lesions in several areas of the near-infrared spectrum. In addition, significant differences were found between the lesion groups by analysis of variance. Linear discriminant analysis classified spectra from benign lesions compared with premalignant or malignant lesions with high accuracy. Near-infrared spectroscopy is a promising noninvasive technique for the screening of skin lesions.

In vivo optical/near-infrared spectroscopy and imaging of metalloproteins.

A number of medical applications of near-infrared spectroscopy are growing closer to clinical acceptance, and new techniques involving both spectroscopy and imaging are evolving rapidly. In vivo spectroscopy and, more recently, imaging techniques are largely based upon optical electronic transitions involving the metal centers of hemoglobin (blood), myoglobin (muscle) and cytochrome aa3 (mitochondria). The wide variety of near-IR based applications includes heart and stroke research, monitoring cerebral oxygenation of premature babies, and 'functional activation' (response of brain to mental tasks). All of these applications are founded upon changes in hemoglobin O2 saturation; these changes are monitored by following trends in the near-infrared absorptions of deoxyhemoglobin (760 nm) and oxyhemoglobin (920 nm). The same absorptions provide a basis for imaging regional variations in blood oxygenation. This report presents and discusses examples, both from the literature and from our recent work, of near-infrared spectroscopy and imaging in medical applications.

Classification of breast tumors by grade and steroid receptor status using pattern recognition analysis of infrared spectra.

Infrared (IR) spectroscopy applied to tissue sections yields complex spectra that provide a molecular fingerprint of the tissue. We have studied a cohort of 77 breast tumors by IR spectroscopy to develop an objective method for the assignment of grade of breast tumors. Although the major variations between spectra from different tumors were in absorptions arising from triglycerides (adipose tissue) and collagen, subtle changes in spectra could be detected that were independent of cellularity and tissue composition. Using a specific multivariate pattern recognition strategy to associate these changes in spectra with different tumor grades, we then were able to accurately reclassify tumors by grade (87% accuracy; kappa = 0.835). A similar approach allowed classification of steroid receptor status (93% accuracy; kappa = 0.852). We conclude that IR spectroscopy may have clinical utility in the objective assignment of breast tumor grade.

Tissue viability by multispectral near infrared imaging: a fuzzy C-means clustering analysis.

Clinically, skin color, temperature, and capillary perfusion are used to assess tissue viability following microvascular tissue transfer. However, clinical signs that arise as a consequence of poor perfusion become evident only after several hours of compromised perfusion. This study demonstrates the potential usefulness of optical/infrared multispectral imaging in the prognosis of tissue viability immediately post-surgery. Multispectral images of a skin flap model acquired within 1 h of surgical elevation are analyzed in comparison to the final 72-h clinical outcome with a high degree of correlation. Regional changes in tissue perfusion and oxygenation present immediately following surgery are differentiated using fuzzy clustering and image processing algorithms. These methodologies reduce the intersubject variability inherent in infrared imaging methods such that the changes in perfusion are reproducible and clearly distinguishable across all subjects. Clinically, an early prognostic indicator of viability such as this would allow for a more timely intervention following surgery in the event of compromised microvasculature.

Fuzzy C-means clustering and principal component analysis of time series from near-infrared imaging of forearm ischemia.

Fuzzy C-means clustering and principal components analysis were used to analyze a temporal series of near-IR images taken of a human forearm during periods of venous outflow restriction and complete forearm ischemia. The principal component eigen-time course analysis provided no useful information and the principal component eigen-image analysis gave results that correlated poorly with anatomical features. The fuzzy C-means clustering analysis, on the other hand, showed distinct regional differences in the hemodynamic response and scattering properties of the tissue, which correlated well with the anatomical features of the forearm.

Placebo-controlled, blind study of dietary manipulation therapy in rheumatoid arthritis.

In a blind, placebo-controlled study of dietary manipulation therapy in outpatients with rheumatoid arthritis there was significant objective improvement during periods of dietary therapy compared with periods of placebo treatment, particularly among "good responders". Possible explanations for improvement include reduced food intolerance, reduced gastrointestinal permeability, and benefit from weight loss and from altered intake of substrates for prostaglandin production. A proportion of the improvement was due to a placebo response, but this was not sufficient to explain the whole improvement.