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In post-stroke hemiparesis, neural impairment alters muscle control, causing abnormal movement and posture in the affected limbs. A decrease in voluntary use of the paretic arm and flexed posture during rest also induce secondary tissue transformation in the upper limb muscles. To obtain a specific, accurate, and reproducible marker of the current biological status of muscles, we collected visible (VIS) and short-wave Infrared (SWIR) reflectance spectra in vivo using a portable spectroradiometer (350–2500 nm), which provided the spectral fingerprints of the elbow flexors and extensors. We compared the spectra for the affected and unaffected sides in 23 patients with post-stroke hemiparesis (25–87 years, 8 women) and eight healthy controls (33–87 years, 5 women). In eight patients, spectra were collected before and after botulinum toxin injection. Spectra underwent off-line preprocessing, principal component analysis, and partial least-squares discriminant analysis. Spectral fingerprints discriminated the muscle (biceps vs. triceps), neurological condition (normal vs. affected vs. unaffected), and effect of botulinum toxin treatment (before vs. 30 to 40 days vs. 110 to 120 days after injection). VIS-SWIR spectroscopy proved valuable for non-invasive assessment of optical properties in muscles, enabled more comprehensive evaluation of hemiparetic muscles, and provided optimal monitoring of the effectiveness of medication.

Recent advances in materials and fabrication techniques provided portable, performant, sensing optical spectrometers readily operated by user-friendly cabled or wireless systems. Such systems allow rapid, non-invasive, and not destructive quantitative analysis of human tissues. This proof-of-principle investigation tested whether infrared spectroscopy techniques, currently utilized in a variety of areas, could be applied in living humans to categorize muscles. Using an ASD FieldSpec® 4 Standard-Res Spectroradiometer with a spectral sampling capability of 1.4 nm at 350–1000 nm and 1.1 nm at 1001–2500 nm, we acquired reflectance spectra in visible short-wave infra-red regions (350–2500 nm) from the upper limb muscles (flexors and extensors) of 20 healthy subjects (age 25–89 years, 9 women). Spectra off-line analysis included preliminary preprocessing, Principal Component Analysis, and Partial Least-Squares Discriminant Analysis. Near-infrared (NIR) spectroscopy proved valuable for noninvasive assessment of tissue optical properties in vivo. In addition to the non-invasive detection of tissue oxygenation, NIR spectroscopy provided the spectral signatures (ie, “fingerprints”) of upper limb flexors and extensors, which represent specific, accurate, and reproducible measures of the overall biological status of these muscles. Thus, non-invasive NIR spectroscopy enables more thorough evaluation of the muscular system and optimal monitoring of the effectiveness of therapeutic or rehabilitative interventions.

Human skeletal muscles may undergo qualitative and quantitative, physiological and pathological changes during life. Some of these changes may be detected with imaging techniques, others with immunohystochemical and molecular analysis. Both these types of investigation are expensive, time consuming, and not readily available. Therefore, at present, a cheap, reliable, and widely applicable technique for non-invasive in vivo analysis of human muscles is lacking. Here we propose optical spectroscopy as a tool that can be added to clinical investigation without significant cost and time penalties. Recent advances in materials and fabrication techniques provided portable, performant, sensing optical spectrometers readily operated by user-friendly cabled or wireless systems. Such systems allow rapid, non-invasive and not destructive quantitative analysis of human tissues. In this investigation, we tested whether infrared spectroscopy techniques, currently utilized in many areas as primary/secondary raw materials sector, cultural heritage, agricultural/food industry, environmental remote and proximal sensing, pharmaceutical industry, etc., could be applied in living humans to categorize muscles. We acquired muscles reflectance spectra in the Vis-SWIR regions (350-2500 nm), utilizing an ASD FieldSpec 4™ Standard-Res Spectroradiometer with a spectral sampling capability of 1.4 nm at 350- 1000 nm and 1.1 nm at 1001-2500 nm. Spectra were collected from the upper limb muscles (i.e. biceps, a forearm flexor, and triceps, a forearm extensor) placed in fixed limb postures (elbow angle approximately 90°) of 22 healthy subjects (age 25-89 years, 11 females). Spectra off-line analysis included preliminary pre-processing for signal scattering reduction, Principal Component Analysis (PCA) aimed to spectral grouping and Partial Least-Squares Discriminant Analysis (PLS-DA) for implementing discrimination/prediction models. Spectral data were correlated with anthropometric variables. Optical spectroscopy proved effective for studying human muscles in vivo. Vis-SWIR spectra acquired from the arm detect muscles from other tissues, and distinguished flexors from extensors.