The first response to infection in the blood is mediated by leukocytes. As a result crucial information can be gained from hemogram. Conventional methods such as smears and automated sorting procedures are not capable of recording detailed biochemical different In this study, Raman spectroscopy has been applied investigate differences between leukocyte subtypes which have obtained healthy donors. imaging was able visualize same morphological features standard staining without need any label....

Rapid and effective methods of pathogen identifications are major interest in clinical microbiological analysis to administer timely tailored antibiotic therapy. Raman spectroscopy as a label-free, culture-independent optical method is suitable identify even single bacteria. However, the low bacteria concentration body fluids makes it difficult detect their characteristic molecular fingerprint directly suspension. Therefore, this study, combined with dielectrophoresis, which enables direct...

We present a high-throughput screening Raman spectroscopy (HTS-RS) platform for rapid and label-free macromolecular fingerprinting of tens thousands eukaryotic cells. The newly proposed HTS-RS combines automated imaging microscopy with to enable cells can be applied large number biomedical clinical applications. potential the new approach is illustrated by two (1) HTS-RS-based differential white blood cell count. A classification model was trained using spectra 52 218 lymphocytes, 48 220...

An experimental evaluation of the information content two complimentary techniques, linear Raman and coherent anti-Stokes scattering (CARS) microscopy, is presented. CARS a nonlinear variant spectroscopy that enables rapid acquisition images within seconds in combination with laser scanning microscopes. were recorded from thin colon tissue sections at 2850, 1660, 1450 1000 cm(-1) compared images. obtained univariate multivariate (k-means clustering) methods, whereas all represent results....

Neutrophils are important cells of the innate immune system and major leukocyte subpopulation in blood. They responsible for recognizing neutralizing invading pathogens, such as bacteria or fungi. For this, neutrophils well equipped with pathogen receptors, cytokines, effector molecules, granules filled reactive oxygen species (ROS)-producing enzymes. Depending on type, different reactions triggered, which result specific activation states neutrophils. Here, we aim to establish a label-free...

Abstract The intestinal epithelial barrier, among other compartments such as the mucosal immune system, contributes to maintenance of homeostasis. Therefore, any disturbance within layer could lead permeability and promote inflammation. Considering that disintegration barrier is a key element in etiology ulcerative colitis, further assessment integrity contribute better understanding role defects colitis (UC), one major form chronic inflammatory bowel disease. Herein, we employ fast,...

For Raman spectroscopic analyses of the cells and other biological samples, choice right substrate material is very important to avoid loss information in characteristic spectral features because competing background signals. In current study, spectroscopy used characterize several potential substrates. vibrational bands are discussed. The surface topography analyzed by atomic force microscopy, root mean square roughness values reported. Biocompatibility substrates tested with Hep G2...

Biofilms are microbial aggregates of microorganisms surrounded by a hydrogel-like matrix formed extracellular polymeric substances (EPS). The formation biofilms is intrinsically complex, from the attachment cells to dispersion biofilm. Meanwhile, three-dimensional framework built up EPS changes with time and protects against environmental stress. Simultaneously acquiring chemical structural information within biofilm vital for cognition regulation biofilms, yet it remains great challenge due...

Dye-loaded micelles of 10 nm diameter formed from amphiphilic graft copolymers composed a hydrophobic poly(methyl methacrylate) backbone and hydrophilic poly(2-ethyl-2-oxazoline) side chains with degree polymerization 15 were investigated concerning their cellular interaction uptake in vitro as well local circulating cells the reticuloendothelial system liver by intravital microscopy. Despite high molar mass individual macromolecules (Mn ≈ 20 kg mol-1), end group modification attachment...

Long-term stability of Raman setups is one the critical criteria for using spectroscopy in real-world applications. Substantial differences from long-term drifts a device can largely reduce the...

Abstract Strategies to deliver drugs using nanocarriers, which are passively or actively targeted their alleged site of action might favorably affect benefit–risk profiles novel therapeutics. Here we tested the hypothesis whether physico-chemical properties cargo as well actual conditions during encapsulation interfere formulation nanoparticular cargo–carrier systems. On basis previous work, a versatile class nanocarriers is polyether-based ABC triblock terpolymer micelles with diameters...

The classification of leukocyte subtypes is a routine method to diagnose many diseases, infections, and inflammations.By applying an automated cell counting procedure, it possible decrease analysis time increase the number analyzed cells per patient, thereby making more robust.Here we propose method, which automatically differentiate between two white blood subtypes, are present in highest fractions.We apply generalized pseudo-Zernike moments transfer morphological information features...

Abstract T lymphocytes (T cells) are highly specialized members of the adaptive immune system and hold key to understanding hosts’ response toward invading pathogen or pathogen-associated molecular patterns such as LPS. In this study, noninvasive Raman spectroscopy is presented a label-free method follow LPS-induced changes in splenic cells during acute postacute inflammatory phases (1, 4, 10, 30 d) with special focus on CD4+ CD8+ endotoxemic C57BL/6 mice. spectral analysis reveals highest...

The human innate immune system is able to recognize pathogen-associated molecular patterns like lipopolysaccharides (LPS) leading the activation of signal cascades and release different cytokines. Activation cells can be assessed in ways which are either indirect (ELISA cytokine release), require staining protocols (flow cytometry) or lysis (mRNA analysis). Here, Raman spectroscopy as a non-destructive spectroscopic method presented enable direct label-free monitoring changes cellular...

Objectives: Leukocytes are first responders to infection. Their activation state can reveal information about specific host immune response and identify dysregulation in sepsis. This study aims use the Raman spectroscopic fingerprints of blood-derived leukocytes differentiate inflammation, infection, sepsis hospitalized patients. Diagnostic sensitivity specificity shall demonstrate added value direct characterization leukocyte’s phenotype. Design: Prospective nonrandomized, single-center,...

In hemolytic disorders, erythrocyte lysis results in massive release of hemoglobin and, subsequently, toxic heme. Hemopexin is the major protective factor against heme toxicity human blood and currently considered for therapeutic use. It has been widely accepted that hemopexin binds with extraordinarily high affinity <1 pM a 1:1 ratio. However, several lines evidence point to higher stoichiometry lower than determined 50 years ago. Here, we re-analyzed these data. SPR UV/Vis spectroscopy...

Multidirectional imaging flow cytometry (mIFC) extends conventional (IFC) for the image-based measurement of 3D-geometrical features particles. The innovative core is a rotation unit in which vertical sample lamella incrementally rotated by 90 degrees into horizontal lamella. required multidirectional views are generated guiding all particles at controllable shear position parabolic velocity profile capillary slit detection chamber. All pass chamber two-dimensional sheet under controlled...

Biochemical information from activated leukocytes provide valuable diagnostic information. In this study, Raman spectroscopy was applied as a label-free analytical technique to characterize the activation pattern of leukocyte subpopulations in an vitro infection model. Neutrophils, monocytes, and lymphocytes were isolated healthy volunteers stimulated with heat-inactivated clinical isolates Candida albicans, Staphylococcus aureus, Klebsiella pneumoniae. Binary classification models could...

Targeted delivery of oligonucleotides or small molecular drugs to hepatocytes, the liver's parenchymal cells, is challenging without targeting moiety due highly efficient mononuclear phagocyte system (MPS) liver. The MPS comprises Kupffer cells and specialized sinusoidal endothelial efficiently clearing nanocarriers regardless their size surface properties. Physiologically, this non-parenchymal shield protects hepatocytes; however, these local barriers must be overcome for drug delivery....

Major drawbacks of direct mid-infrared spectroscopic imaging single cells in an aqueous buffer are strong water absorption, low resolution typically above 10 μm, and Mie scattering effects. This study demonstrates how indirect detection principle can overcome these using the optical photothermal infrared (O-PTIR) technique for high-resolution discrete wavenumber fingerprint spectroscopy cultivated as a model system simple liquid sample chamber. The O-PTIR spectra six leukemia- cancer-derived...