Abstract
The significance and utility of innovative imaging techniques in arterial clot analysis, which enable far more detailed and automated analysis compared to standard methods, are presented. The examination of two types of human thrombi is shown, representing the main ischemic stroke etiologies: fibrin–predominant clot of large vessel origin and red blood cells–rich clot of cardioembolic origin. The synergy effect of Fourier–transform infrared spectroscopy (FTIR), Raman spectroscopy (RS) and atomic force microscopy (AFM) techniques supported by chemometrics in comparison with reference histological staining was presented. The main advantage of such approach refers to free–label and non–destructive quantitative imaging of clinically valid, biochemical parameters in whole sample (FTIR–low resolution) and selected regions (RS–ultra–high resolution). We may include here analysis of lipid content, its distribution and total degree of unsaturation as well as analysis of protein content (mainly fibrin and hemoproteins). The AFM studies enhanced the vibrational data, showed clearly shape and thickness of clot features as well as visualized the fibrin framework. The extraordinary sensitivity of FTIR and RS imaging toward detection and discrimination of clinically valid parameters in clot confirms its applicability in assessment of thrombi origin.
Highlights
Stroke is the fifth cause of death globally accounting 6.5 millions of deaths worldwide[1]
The results section is divided into two parts which presents the analysis of thrombi which were representative for a different etiology of the acute ischemic stroke: one sample was derived from a patient suffered from large vessel stroke due to artery dissection while two other samples were collected from two patients with cardioembolic stroke
Different parts of thrombus reveal a diversity of histological www.nature.com/scientificreports pattern, wherein fibrin is stained in pink and is present as a homogenous area localized on the edges and in the middle of the clot
Summary
Stroke is the fifth cause of death globally accounting 6.5 millions of deaths worldwide[1]. Computed tomography (CT) and magnetic resonance imaging (MRI) studies have been correlated previously with the clot histological composition to assess thrombolytic recanalization outcome and IS etiology. On contrary a number of studies have reported that CT and MRI give poor prognostic information on clinical outcome and have a limited clinical diagnostic value[4,7,8] It is a heterogeneous disease with numerous underlying causes. The frozen cross–sections of the retrieved thrombi were firstly examined with free–label FTIR imaging which allowed us to study the distribution of various clot features with the spatial resolution of around 5.5 μm[14]. The main goal of this work was to present the benefits of the combination of FTIR, RS and AFM techniques for the thrombi analysis and to highlight the potential of such assessment to identify brain clot origin – crucial for future stroke prevention. To the best of our knowledge this is the first study to utilize label–free and chemically sensitive imaging of acute IS retrieved clot
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