Spectroscopic Optical Tomography Research Articles

Near-infrared evolution of the equatorial ring of SN 1987A

We used adaptive optics imaging and integral field spectroscopy from the Very Large Telescope, together with images from the Hubble Space Telescope, to study the near-infrared (NIR) evolution of the equatorial ring (ER) of SN 1987A. We studied the NIR flux and morphology over time in order to lay the groundwork for James Webb Space Telescope observations of the system. We also studied the differences in the interacting ring structure and flux between optical, NIR, and other wavelengths, and between line and continuum emission, to constrain the underlying physical processes. For the most part, the evolution is similar in the NIR and optical. The morphology of the ER has been skewed toward the west side (with roughly two-thirds of the NIR emission originating there) since around 2010. A steady decline in the ER flux, broadly similar to the mid-infrared and the optical, has been ongoing since roughly this time as well. The expansion velocity of the ER hotspots in the NIR is fully consistent with the optical. However, continuum emission forms roughly 70% of the NIR luminosity, and has been stronger outside the hotspot-defined extent of the ER (relative to the hotspots themselves) than the optical emission or the NIR line emission since 2012–2013, suggesting a faster-expanding continuum component. We find that this outer NIR emission can have a significant synchrotron contribution. Even if emission from hot dust (~2000 K) is dominant within the ER, the mass of this dust must be vanishingly small (a few times 10−12 M⊙) compared to the total dust mass in the ER (≳10−5 M⊙) to account for the observed HKs flux. The NIR continuum emission, however, expands more slowly than the more diffuse 180-K dust emission that dominates in the MIR, indicating a different source, and the same hot dust component cannot account for the J-band emission.

Changes in the tumor oxygenation but not in the tumor volume and tumor vascularization reflect early response of breast cancer to neoadjuvant chemotherapy

BackgroundBreast cancer neoadjuvant chemotherapy (NACT) allows for assessing tumor sensitivity to systemic treatment, planning adjuvant treatment and follow-up. However, a sufficiently large number of patients fail to achieve the desired level of pathological tumor response while optimal early response assessment methods have not been established now. In our study, we simultaneously assessed the early chemotherapy-induced changes in the tumor volume by ultrasound (US), the tumor oxygenation by diffuse optical spectroscopy imaging (DOSI), and the state of the tumor vascular bed by Doppler US to elaborate the predictive criteria of breast tumor response to treatment.MethodsA total of 133 patients with a confirmed diagnosis of invasive breast cancer stage II to III admitted to NACT following definitive breast surgery were enrolled, of those 103 were included in the final analysis. Tumor oxygenation by DOSI, tumor volume by US, and tumor vascularization by Doppler US were determined before the first and second cycle of NACT. After NACT completion, patients underwent surgery followed by pathological examination and assessment of the pathological tumor response. On the basis of these, data regression predictive models were created.ResultsWe observed changes in all three parameters 3 weeks after the start of the treatment. However, a high predictive potential for early assessment of tumor sensitivity to NACT demonstrated only the level of oxygenation, ΔStO2, (ρ = 0.802, p ≤ 0.01). The regression model predicts the tumor response with a high probability of a correct conclusion (89.3%). The “Tumor volume” model and the “Vascularization index” model did not accurately predict the absence of a pathological tumor response to treatment (60.9% and 58.7%, respectively), while predicting a positive response to treatment was relatively better (78.9% and 75.4%, respectively).ConclusionsDiffuse optical spectroscopy imaging appeared to be a robust tool for early predicting breast cancer response to chemotherapy. It may help identify patients who need additional molecular genetic study of the tumor in order to find the source of resistance to treatment, as well as to correct the treatment regimen.

How does alzheimer’s disease, atherosclerosis and comorbid alzheimer’s disease and atherosclerosis impact neurovascular function?

AbstractBackgroundThe brain is an energy hungry organ, consuming approximately 20% of the body’s energy supply (Clark & Sokoloff 1999). Due to its high energy demands the brain has developed a critical mechanism termed neurovascular coupling (NVC). NVC is the relationship between neuronal activity and a subsequent increase in blood flow to the same brain region. This mechanism is suggested to be impaired in many disorders, including Alzheimer’s disease (AD) (Kisler et al., 2017). Additionally, approximately 90% of individuals living with dementia have a comorbidity (Browne et al., 2017), with some of the most common comorbidities being cardiovascular disease (Tini et al., 2020) and type 2 diabetes (Santiago et al., 2021). Therefore, we wanted to investigate neurovascular function in AD and comorbid mixed models of AD and atherosclerosis.MethodFour separate groups of mice were used, aged 9‐12 months: AD (APP/PS1 (B6;C3 Tg(APPswe,PSEN1dE9)85Dbo/Mmjax)), WT (APP/PS1‐WT littermates) , atherosclerosis (WT + rAAV8‐mPCSK9‐D377Y + Western Diet) and Mixed AD and atherosclerosis (APP/PS1 + rAAV8‐mPCSK9‐D377Y + Western Diet). At 9m mice completed the novel object recognition test (NOR) to assess cognition. Following this, a thinned window surgery was completed. One‐week post‐surgery 2‐dimensional optical imaging spectroscopy (2D‐OIS) was utilised to investigate changes in cerebral haemodynamics, in response to a whisker stimulation in awake mice. A separate imaging session in anaesthetised mice was also completed, using 2D‐OIS concurrent with electrophysiology to assess neurovascular coupling. Immunohistochemistry was performed to assess amyloid plaque load within the brain.ResultIn the awake imaging session we found that neurovascular function was preserved in the AD group. However, mice in the atherosclerosis group showed a reduction in blood volume in response to a 2s‐whisker stimulation, compared to the WT group. We hypothesised that the mixed model would have the greatest impairment in the neurovascular response, however this was not observed and this group showed an evoked‐blood volume change in between levels of AD and atherosclerosis only groups.ConclusionPreliminary results suggest that neurovascular function is impaired in mouse models of atherosclerosis but not in the AD or mixed models. Analysis is ongoing to determine how neural activity and AD pathology are impacted by comorbidity.

Prognostic potential and pathological validation of a diagnostic application using Raman spectroscopy in the characterization of degenerative changes in the cartilage of the humeral head.

Raman spectroscopy is a well-established analytical method in the fields of chemistry, industry, biology, pharmaceutics, and medicine. Previous studies have investigated optical imaging and Raman spectroscopy for osteoarthritis (OA) diagnosis in weight-bearing joints such as hip and knee joints. However, to realize early diagnosis or a curable treatment, it is still challenging to understand the correlations with intrinsic factors or patients’ background. To elucidate the correlation between the Raman spectral features and pathological variations of human shoulder joint cartilage. Osteoarthritic cartilage specimens excised from the humeral heads of 14 patients who underwent shoulder arthroplasty were assessed by a confocal Raman microscope and histological staining. The Raman spectroscopic dataset of degenerative cartilage was further analyzed by principal component analysis and hierarchical cluster analysis. Multivariate association of the Raman spectral data generated three major clusters. The first cluster of patients shows a relatively high Raman intensity of collagen. The second cluster displays relatively low Raman intensities of proteoglycans (PGs) and glycosaminoglycans (GAGs), whereas the third cluster shows relatively high Raman intensities of PGs and GAGs. The reduced PGs and GAGs are typical changes in OA cartilage, which have been confirmed by safranin–O staining. In contrast, the increased Raman intensities of collagen, PGs, and GAGs may reflect the instability of the cartilage matrix structure in OA patients. The results obtained confirm the correlation between the Raman spectral features and pathological variations of human shoulder joint cartilage. Unsupervised machine learning methods successfully yielded a clinically meaningful classification between the shoulder OA patients. This approach not only has potential to confirm severity of cartilage defects but also to determine the origin of an individual’s OA by evaluating the cartilage quality.

Optical imaging spectroscopy coupled with machine learning for detecting heavy metal of plants: A review.

Heavy metal elements, which inhibit plant development by destroying cell structure and wilting leaves, are easily absorbed by plants and eventually threaten human health via the food chain. Recently, with the increasing precision and refinement of optical instruments, optical imaging spectroscopy has gradually been applied to the detection and reaction of heavy metals in plants due to its in-situ, real-time, and simple operation compared with traditional chemical analysis methods. Moreover, the emergence of machine learning helps improve detection accuracy, making optical imaging spectroscopy comparable to conventional chemical analysis methods in some situations. This review (a): summarizes the progress of advanced optical imaging spectroscopy techniques coupled with artificial neural network algorithms for plant heavy metal detection over ten years from 2012-2022; (b) briefly describes and compares the principles and characteristics of spectroscopy and traditional chemical techniques applied to plants heavy metal detection, and the advantages of artificial neural network techniques including machine learning and deep learning techniques in combination with spectroscopy; (c) proposes the solutions such as coupling with other analytical and detection methods, portability, to address the challenges of unsatisfactory sensitivity of optical imaging spectroscopy and expensive instruments.

In Situ FTIR Spectroscopic Imaging of Asphaltene Deposition from Crude Oil under n-Heptane and Acetone Flows

Asphaltene deposition from crude oil, induced by n-heptane and acetone flows in a microfluidic device, was studied by optical and FTIR spectroscopic imaging techniques in situ. It was found that n-heptane and acetone penetrate crude oil in completely different ways, which is responsible for the formation of deposits with different structures and chemical compositions. The contact of the n-heptane flow with crude oil leads to fast aggregation of asphaltenes at the interface, resulting in the formation of a compact deposit fixed to the surface. Subsequent slow diffusion of n-heptane into crude oil causes asphaltene aggregation and precipitation of loose deposits. In the case of acetone, the flocculant diffuses (faster than n-heptane) into crude oil, which results in removal of its soluble components and in the formation of deposits appearing as strips as well. The distribution of the functional groups (–OH(NH), CH2–CH3, C=O, C–O, S=O) in the deposits is spatially heterogeneous. The asphaltenes deposits formed at the flocculant-crude oil interface are richer in the functional groups than those formed during diffusion. No carbonyl groups were revealed in the deposits formed in the acetone flow, while oxygen-containing groups such as –OH(NH) and S=O were present, which was an unusual result.

Deep UV Led Dynamic Optical Imaging and Fluorescence Spectroscopy of the Protein Corona in a Plasmonic Solution and the Effect of Near-Infrared Laser Heating

Deep UV Led Dynamic Optical Imaging and Fluorescence Spectroscopy of the Protein Corona in a Plasmonic Solution and the Effect of Near-Infrared Laser Heating

The Bow Shock and Kinematics of PSR J1959+2048

Pulsar Hα bow shocks provide rare opportunities to constrain the energetics and kinematics of the relativistic pulsar wind. We have acquired optical imaging and integral field unit spectroscopy of the bow shock of the millisecond pulsar PSR J1959+2048, measuring the shock symmetry axis at a position angle = 213.2 ± 0.°2 and showing that this slow nonradiative shock has a broad-to-narrow line component ratio I b /I n = 4. The data show that the pulsar’s velocity lies 2.°2 out of the plane of the sky. Coupled with a new fit for its timing proper motion, giving μ tot = 30.05 mas yr−1 and a Very Long Baseline Array (VLBA) interferometric parallax measurement giving kpc (90% range), we have unusually complete information on the pulsar kinematics. The bow shock constraints on the wind momentum flux imply that, at the best-fit parallax distance, the pulsar moment of inertia must be very large and/or the Hα efficiency at its modest shock velocity must be very high.

Optical imaging spectroscopy for rapid, primary screening of SARS-CoV-2: a proof of concept

Effective testing is essential to control the coronavirus disease 2019 (COVID-19) transmission. Here we report a-proof-of-concept study on hyperspectral image analysis in the visible and near-infrared range for primary screening at the point-of-care of SARS-CoV-2. We apply spectral feature descriptors, partial least square-discriminant analysis, and artificial intelligence to extract information from optical diffuse reflectance measurements from 5 µL fluid samples at pixel, droplet, and patient levels. We discern preparations of engineered lentiviral particles pseudotyped with the spike protein of the SARS-CoV-2 from those with the G protein of the vesicular stomatitis virus in saline solution and artificial saliva. We report a quantitative analysis of 72 samples of nasopharyngeal exudate in a range of SARS-CoV-2 viral loads, and a descriptive study of another 32 fresh human saliva samples. Sensitivity for classification of exudates was 100% with peak specificity of 87.5% for discernment from PCR-negative but symptomatic cases. Proposed technology is reagent-free, fast, and scalable, and could substantially reduce the number of molecular tests currently required for COVID-19 mass screening strategies even in resource-limited settings.

Design and Construction of an Optical TEM Specimen Holder

Abstract:Electron microscopy has enabled atomic resolution imaging of matter. However, unlike optical spectroscopic imaging, traditional electron microscopes provide limited spectroscopic information in terms of their energy resolution. Only recently, owing to advances in monochromated STEM-EELS, have transmission electron microscopes (TEMs) been able to attain a high energy resolution. We recently proposed combining spectrally selective photoexcitation with HRTEM to achieve sub-nanometer scale optical imaging, a technique we called photoabsorption microscopy using electron analysis (PAMELA). To realize PAMELA-TEM experimentally, we constructed a TEM holder with an optical feedthrough, capable of photoexciting materials with different wavelengths. In this article, we describe our process for designing and fabricating an optical TEM specimen holder, highlighting important aspects of the design.

Multi-modal diffuse optical spectroscopy for high-speed monitoring and wide-area mapping of tissue optical properties and hemodynamics.

.Significance: Diffuse optical spectroscopic imaging (DOSI) is a versatile technology sensitive to changes in tissue composition and hemodynamics and has been used for a wide variety of clinical applications. Specific applications have prompted the development of versions of the DOSI technology to fit specific clinical needs. This work describes the development and characterization of a multi-modal DOSI (MM-DOSI) system that can acquire metabolic, compositional, and pulsatile information at multiple penetration depths in a single hardware platform. Additionally, a 3D tracking system is integrated with MM-DOSI, which enables registration of the acquired data to the physical imaging area.Aim: We demonstrate imaging, layered compositional analysis, and metabolism tracking capabilities using a single MM-DOSI system on optical phantoms as well as in vivo human tissue.Approach: We characterize system performance with a silicone phantom containing an embedded object. To demonstrate multi-layer sensitivity, we imaged human calf tissue with a 4.8-mm skin-adipose thickness. Human thenar tissue was also measured using a combined broadband DOSI and continuous-wave near-infrared spectroscopy method ( acquisition rate).Results: High-resolution optical property maps of absorption () and reduced scattering () were recovered on the phantom by capturing over 1000 measurement points in under 5 minutes. On human calf tissue, we show two probing depth layers have significantly different () total-hemo/myoglobin and composition. On thenar tissue, we calculate tissue arterial oxygen saturation, venous oxygen saturation, and tissue metabolic rate of oxygen consumption during baseline and after release of an arterial occlusion.Conclusions: The MM-DOSI can switch between collection of broadband spectra, high-resolution images, or multi-depth hemodynamics without any hardware reconfiguration. We conclude that MM-DOSI enables acquisition of high resolution, multi-modal data consolidated in a single platform, which can provide a more comprehensive understanding of tissue hemodynamics and composition for a wide range of clinical applications.

Highly Efficient Anisotropic Chiral Plasmonic Metamaterials for Polarization Conversion and Detection.

Plasmonic chiral metamaterials have attracted broad research interest because of their potential applications in optical communication, biomedical diagnosis, polarization imaging, and circular dichroism spectroscopy. However, optical losses in plasmonic structures severely limit practical applications. Here, we present the design concept and experimental demonstration for highly efficient subwavelength-thick plasmonic chiral metamaterials with strong chirality. The proposed designs utilize plasmonic metasurfaces to control the phase and polarization of light and exploit anisotropic thin-film interference effects to enhance optical chirality while minimizing optical loss. Based on such design concepts, we demonstrated experimentally optical devices such as circular polarization filters with transmission efficiency up to 90% and extinction ratio >180, polarization converters with conversion efficiency up to 90%, as well as on-chip integrated microfilter arrays for full Stokes polarization detection with high accuracy over a broad wavelength range (3.5-5 μm). The proposed design concepts are applicable from near-infrared to Terahertz regions via structural engineering.

Glyceryl trinitrate for the treatment of ischaemic stroke: Determining efficacy in rodent and ovine species for enhanced clinical translation

Hypertension is a leading risk factor for death and dependency after ischaemic stroke. However, administering anti-hypertensive medications post-stroke remains contentious with concerns regarding deleterious effects on cerebral blood flow and infarct expansion. This study sought to determine the effect of glyceryl trinitrate (GTN) treatment in both lissencephalic and gyrencephalic pre-clinical stroke models. Merino sheep underwent middle cerebral artery occlusion (MCAO) followed by GTN or control patch administration (0.2 mg/h). Monitoring of numerous physiologically relevant measures over 24 h showed that GTN administration was associated with decreased intracranial pressure, infarct volume, cerebral oedema and midline shift compared to vehicle treatment (p < 0.05). No significant changes in blood pressure or cerebral perfusion pressure were observed. Using optical imaging spectroscopy and laser speckle imaging, the effect of varying doses of GTN (0.69–50 µg/h) on cerebral blood flow and tissue oxygenation was examined in mice. No consistent effect was found. Additional mice undergoing MCAO followed by GTN administration (doses varying from 0–60 µg/h) also showed no improvement in infarct volume or neurological score within 24 h post-stroke. GTN administration significantly improved numerous stroke-related physiological outcomes in sheep but was ineffective in mice. This suggests that, whilst GTN administration could potentially benefit patients, further research into mechanisms of action are required.

Validating layer-specific VASO across species

Cerebral blood volume (CBV) has been shown to be a robust and important physiological parameter for quantitative interpretation of functional (f)MRI, capable of delivering highly localized mapping of neural activity. Indeed, with recent advances in ultra-high-field (≥7T) MRI hardware and associated sequence libraries, it has become possible to capture non-invasive CBV weighted fMRI signals across cortical layers. One of the most widely used approaches to achieve this (in humans) is through vascular-space-occupancy (VASO) fMRI. Unfortunately, the exact contrast mechanisms of layer-dependent VASO fMRI have not been validated for human fMRI and thus interpretation of such data is confounded. Here we validate the signal source of layer-dependent SS-SI VASO fMRI using multi-modal imaging in a rat model in response to neuronal activation (somatosensory cortex) and respiratory challenge (hypercapnia). In particular VASO derived CBV measures are directly compared to concurrent measures of total haemoglobin changes from high resolution intrinsic optical imaging spectroscopy (OIS). Quantified cortical layer profiling is demonstrated to be in agreement between VASO and contrast enhanced fMRI (using monocrystalline iron oxide nanoparticles, MION). Responses show high spatial localisation to layers of cortical processing independent of confounding large draining veins which can hamper BOLD fMRI studies, (depending on slice positioning). Thus, a cross species comparison is enabled using VASO as a common measure. We find increased VASO based CBV reactivity (3.1 ± 1.2 fold increase) in humans compared to rats. Together, our findings confirm that the VASO contrast is indeed a reliable estimate of layer-specific CBV changes. This validation study increases the neuronal interpretability of human layer-dependent VASO fMRI as an appropriate method in neuroscience application studies, in which the presence of large draining intracortical and pial veins limits neuroscientific inference with BOLD fMRI.

Real-Time Handheld Probe Tracking and Image Formation Using Digital Frequency-Domain Diffuse Optical Spectroscopy.

Frequency-domain diffuse optical spectroscopic imaging (FD-DOS) is a non-invasive method for measuring absolute concentrations of tissue chromophores such as oxy- and deoxy-hemoglobin in vivo. The utility of FD-DOS for clinical applications such as monitoring chemotherapy response in breast cancer has previously been demonstrated, but challenges for further clinical translation, such as slow acquisition speed and lack of user feedback, remain. Here, we propose a new high speed FD-DOS instrument that allows users to freely acquire measurements over the tissue surface, and is capable of rapidly imaging large volumes of tissue. We utilize 3D monocular probe tracking combined with custom digital FD-DOS hardware and a high-speed data processing pipeline for the instrument. Results are displayed during scanning over the surface of the sample using a probabilistic Monte Carlo light propagation model. We show this instrument can measure absorption and scattering coefficients with an error of 7% and 1% respectively, with 0.7 mm positional accuracy. We demonstrate the equivalence of our visualization methodology with a standard interpolation approach, and demonstrate two proof-of-concept in vivo results showing superficial vasculature in the human forearm and surface contrast in a healthy human breast. Our new FD-DOS system is able to compute chromophore concentrations in real-time (1.5 Hz) in vivo. This method has the potential to improve the quality of FD-DOS image scans while reducing measurement times for a variety of clinical applications.

Mesoscopic fluorescence lifetime imaging: Fundamental principles, clinical applications and future directions.

Fluorescence lifetime imaging (FLIm) is an optical spectroscopic imaging technique capable of real-time assessments of tissue properties in clinical settings. Label-free FLIm is sensitive to changes in tissue structure and biochemistry resulting from pathological conditions, thus providing optical contrast to identify and monitor the progression of disease. Technical and methodological advances over the last two decades have enabled the development of FLIm instrumentation for real-time, in situ, mesoscopic imaging compatible with standard clinical workflows. Herein, we review the fundamental working principles of mesoscopic FLIm, discuss the technical characteristics of current clinical FLIm instrumentation, highlight the most commonly used analytical methods to interpret fluorescence lifetime data and discuss the recent applications of FLIm in surgical oncology and cardiovascular diagnostics. Finally, we conclude with an outlook on the future directions of clinical FLIm.

Photoabsorption Imaging at Nanometer Scales Using Secondary Electron Analysis.

Optical imaging with nanometer resolution offers fundamental insights into light-matter interactions. Traditional optical techniques are diffraction limited with a spatial resolution >100 nm. Optical super-resolution and cathodoluminescence techniques have higher spatial resolutions, but these approaches require the sample to fluoresce, which many materials lack. Here, we introduce photoabsorption microscopy using electron analysis, which involves spectrally specific photoabsorption that is locally probed using a scanning electron microscope, whereby a photoabsorption-induced surface photovoltage modulates the secondary electron emission. We demonstrate spectrally specific photoabsorption imaging with sub-20 nm spatial resolution using silicon, germanium, and gold nanoparticles. Theoretical analysis and Monte Carlo simulations are used to explain the basic trends of the photoabsorption-induced secondary electron signal. Based on our current experiments and this analysis, we expect that the spatial resolution can be further improved to a few nanometers, thereby offering a general approach for nanometer-scale optical spectroscopic imaging and material characterization.

Breast cancer differential diagnosis using diffuse optical spectroscopic imaging and regression with z-score normalized data.

.Significance: Current imaging paradigms for differential diagnosis of suspicious breast lesions suffer from high false positive rates that force patients to undergo unnecessary biopsies. Diffuse optical spectroscopic imaging (DOSI) noninvasively probes functional hemodynamic and compositional parameters in deep tissue and has been shown to be sensitive to contrast between normal and malignant tissues.Aim: DOSI methods are under investigation as an adjunct to mammography and ultrasound that could reduce false positive rates and unnecessary biopsies, particularly in radiographically dense breasts.Methods: We performed a retrospective analysis of 212 subjects with suspicious breast lesions who underwent DOSI imaging. Physiological tissue parameters were -score normalized to the patient’s contralateral breast tissue and input to univariate logistic regression models to discriminate between malignant tumors and the surrounding normal tissue. The models were then used to differentiate malignant lesions from benign lesions.Results: Models incorporating several individual hemodynamic parameters were able to accurately distinguish malignant tumors from both the surrounding background tissue and benign lesions with area under the curve (AUC) . -score normalization improved the discriminatory ability and calibration of these predictive models relative to unnormalized or ratio-normalized data.Conclusions: Findings from a large subject population study show how DOSI data normalization that accounts for normal tissue heterogeneity and quantitative statistical regression approaches can be combined to improve the ability of DOSI to diagnose malignant lesions. This improved diagnostic accuracy, combined with the modality’s inherent logistical advantages of portability, low cost, and nonionizing radiation, could position DOSI as an effective adjunct modality that could be used to reduce the number of unnecessary invasive biopsies.

Old and new major mergers in the SOSIMPLE galaxy, NGC 7135

ABSTRACT The simultaneous advancement of high resolution integral field unit spectroscopy and robust full-spectral fitting codes now make it possible to examine spatially resolved kinematic, chemical composition, and star-formation history from nearby galaxies. We take new MUSE data from the snapshot optical spectroscopic imaging of mergers and pairs for legacy Exploration (SOSIMPLE) survey to examine NGC 7135. With counter-rotation of gas, disrupted kinematics and asymmetric chemical distribution, NGC 7135 is consistent with an ongoing merger. Though well hidden by the current merger, we are able to distinguish stars originating from an older merger, occurring 6–10 Gyr ago. We further find a gradient in ex-situ material with galactocentric radius, with the accreted fraction rising from 0 per cent in the galaxy centre, to ∼7 per cent within 0.6 effective radii.

Raman spectroscopy as process analytical tool in downstream processing of biotechnology

• Raman spectroscopy can promote the process analytical technologies in bioprocess. • Raman spectroscopy technique is an effective downstream bioprocess monitoring tool. • Chemometric approaches could enrich the sensitivity of Raman spectroscopy. Downstream processing or product recovery plays a vital role in the development of bioprocesses. To improve the bioprocess efficiency, some unconventional methods are much required. The continuous manufacturing in downstream processing makes the Process Analytical Technologies (PATs) as an important tool. Monitoring and controlling bioprocess are an essential factor for the principles of PAT and quality by design. Spectroscopic methods can apply to monitor multiple analytes in real-time with less sample processing with significant advancements. Raman spectroscopy is an extensively used technique as an analytical and research tool owing to its modest process form, non-destructive, non-invasive optical molecular spectroscopic imaging with computer-based analysis. Generally, its application is essential for the analysis and characterization of biological samples, and it is easy to operate with minimal sample. The innovation on various types of enhanced Raman spectroscopy was designed to enhance the Raman analytical technique. Raman spectroscopy could couple with chemometrics to provide reliable alternative analysis method of downstream process analysis. Thus, this review aims to provide useful insight on the application of Raman spectroscopy for PAT in downstream processing of biotechnology and Raman data analysis in biological fields.