scholarly journals Extraction And Quality Evaluation of Exosomes From Joint Effusion

2020 ◽  
Author(s):  
Anmin Ruan ◽  
Pu Chen ◽  
Jun Zhou ◽  
Xiaozhe Zhang ◽  
Yufeng Ma ◽  
...  
Keyword(s):  
Knee Joint ◽  
Particle Morphology ◽  
Extraction Methods ◽  
Joint Effusion ◽  
Precipitation Method ◽  
Transmission Electron ◽  
Marker Proteins ◽  
Exclusion Chromatography ◽  
Peg Precipitation Method

Abstract Objective: The aim of this study was to compare the quality and efficiency of exosomes extracted from knee joint effusion by different methods, laying a foundation for further research on exosomes from knee joint effusion. Methods: Separate and extract joint exosomes by 8% polyethylene glycol (PEG) precipitation method, ultracentrifugation method (UC) and ultrafiltration with exclusion chromatography (SECF). Transmission electron microscopy (TEM) and nanoparticle tracing technology (NTA) were used to detect particle morphology and particle size, and Western Blot (WB) was used to detect granule protein surface marker proteins (CD9, CD63, Flotillin-1 and calnexin).Results: All three methods successfully separated vesicle-like round particles from joint effusion. TEM results show that the particles obtained by the three extraction methods are round or oval vesicles, the plasma membrane is intact, the size is different, and the diameter is distributed between 30 and 150 nm. Compared with SECF group, PEG group had more background particle impurities. and the broken vesicle fragments can be seen in the UC group; NTA results show that the main peaks of the three groups of particles are between 100-120nm, and the particle concentration is greater than 1×1010/ml; WB results show that the expressions of CD9, CD63 and Flotillin-1 protein in the suspension extracted by the three methods were all higher, the expression of calnexin protein was higher in the PEG group than the UC group and SECF group.Conclusion: The three extraction methods can successfully extract the exosomes of joint effusion. The quality of exosomes obtained by the SECF method is relatively high, while the PEG precipitation method contains a small amount of impurity particles. UC method does not guarantee the integrity of exosomes. In summary, when further studying the role of exosomes in the pathogenesis of knee osteoarthritis, we recommend the SECF method to extract and isolate joint exudates.

scholarly journals Extraction And Quality Evaluation of Exosomes From Joint Effusion

2020 ◽  
Author(s):  
Anmin Ruan ◽  
Yueshan Yin ◽  
Pu Chen ◽  
Jun Zhou ◽  
Xiaozhe Zhang ◽  
...  
Keyword(s):  
Knee Joint ◽  
Particle Morphology ◽  
Extraction Methods ◽  
Joint Effusion ◽  
Precipitation Method ◽  
Transmission Electron ◽  
Marker Proteins ◽  
Exclusion Chromatography ◽  
Peg Precipitation Method

Abstract Objective: The aim of this study was to compare the quality and efficiency of exosomes extracted from knee joint effusion by different methods, laying a foundation for further research on exosomes from knee joint effusion. Methods: Separate and extract joint exosomes by 8% polyethylene glycol (PEG) precipitation method, ultracentrifugation method (UC) and ultrafiltration with exclusion chromatography (SECF). Transmission electron microscopy (TEM) and nanoparticle tracing technology (NTA) were used to detect particle morphology and particle size, and Western Blot (WB) was used to detect granule protein surface marker proteins (CD9, CD63, Flotillin-1 and calnexin).Results: All three methods successfully separated vesicle-like round particles from joint effusion. TEM results show that the particles obtained by the three extraction methods are round or oval vesicles, the plasma membrane is intact, the size is different, and the diameter is distributed between 30 and 150 nm. Compared with SECF group, PEG group had more background particle impurities. and the broken vesicle fragments can be seen in the UC group; NTA results show that the main peaks of the three groups of particles are between 100-120nm, and the particle concentration is greater than 1×1010/ml; WB results show that the expressions of CD9, CD63 and Flotillin-1 protein in the suspension extracted by the three methods were all higher, the expression of calnexin protein was higher in the PEG group than the UC group and SECF group.Conclusion: The three extraction methods can successfully extract the exosomes of joint effusion. The quality of exosomes obtained by the SECF method is relatively high, while the PEG precipitation method contains a small amount of impurity particles. UC method does not guarantee the integrity of exosomes. In summary, when further studying the role of exosomes in the pathogenesis of knee osteoarthritis, we recommend the SECF method to extract and isolate joint exudates.

scholarly journals Extraction and quality evaluation of exosomes from joint effusion

2020 ◽  
Author(s):  
Anmin Ruan ◽  
Yueshan Yin ◽  
Pu Chen ◽  
Jun Zhou ◽  
Xiaozhe Zhang ◽  
...  
Keyword(s):  
Knee Joint ◽  
Particle Morphology ◽  
Extraction Methods ◽  
Joint Effusion ◽  
Precipitation Method ◽  
Transmission Electron ◽  
Marker Proteins ◽  
Exclusion Chromatography ◽  
Granule Surface ◽  
Peg Precipitation Method

Abstract Objective: The aim of this study was to compare the quality and efficiency of exosomes extracted from knee joint effusion by different methods, laying a foundation for further research on exosomes from knee joint effusion. Methods:To separate and extract joint exosomes by 8% polyethylene glycol (PEG) precipitation method, ultracentrifugation method (UC) and ultrafiltration with exclusion chromatography (SECF). Transmission electron microscopy (TEM) and nanoparticle tracing technology (NTA) were used to detect particle morphology and particle size, and Western Blot (WB) was used to detect marker proteins of granule surface (CD9, CD63, Flotillin-1 and calnexin).Results: Three methods separated vesicle-like round particles from joint effusion successfully. The results of TEM show that the particles obtained by the three extraction methods are round or oval vesicles, the plasma membrane is intact, the size is different, and the diameter is distributed between 30 and 150 nm. Compared with SECF group, PEG group had more background particle impurities. and the broken vesicle fragments can be seen in the UC group; The results of NTA show that the main peaks of the three groups of particles are between 100-120nm, and the particle concentration is greater than 1×1010/ml; The results of WB show that the protein expressions of CD9, CD63 and Flotillin-1 in the suspension extracted by the three methods were higher, the expression of calnexin protein in the PEG group was higher than the UC group and SECF group.Conclusion: The three extraction methods can extract the exosomes of joint effusion successfully. The quality of exosomes obtained by the SECF method is relatively high, while the PEG precipitation method contains a small amount of impurity particles. UC method does not guarantee the integrity of exosomes. In summary, we recommend the SECF method to extract and isolate joint effusion exudates-derived exosomes when further studying the role of exosomes in the pathogenesis of knee osteoarthritis.

Apoptosis During the Knee Joint Development of Mouse

2000 ◽  
Vol 6 (S2) ◽  
pp. 960-961
Author(s):  
D. H. Hwang ◽  
H. S. Lim ◽  
Y. W. Kim
Keyword(s):  
Knee Joint ◽  
Morphological Characteristics ◽  
Mesenchymal Cells ◽  
Limb Bud ◽  
Tunel Staining ◽  
Developmental Study ◽  
Apoptotic Cells ◽  
Transmission Electron ◽  
Organ Systems

Apoptosis has been known as a significant element in the process of embryonic morphogenesis (1). Many studies on the role of apoptosis during development have been done using several organ systems, such as limb bud (2), nervous system(3), and inner ear (4). Recently, cell death with many morphological characteristics of apoptosis was reported in the developmental study of chick knee joint (5). This study was designed to observe the appearance of apoptotic cells during the joint cavity formation of mouse knee joint by transmission electron microscopy and TUNEL staining. The fetus were collected from l5thday(15D) to 19th day(l 9D) of pregnancy.We observed many blood vessels and mesenchymal cells at 15D between the cartilages of femur and tibia. Mesenchymal cells had condensed heterochromatin and dilated rough endoplasmic reticulum (rER). At 16D, the joint clefts were first formed. Apoptotic cells, with characteristic nuclear fragmentation and condensation, appeared between the joint cleft and the capillaries.

On Future Directions in Pathology

1982 ◽  
Vol 40 ◽  
pp. 274-277
Author(s):  
Benjamin F. Trump ◽  
Irene K. Berezesky ◽  
Raymond T. Jones
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Clinical Pathology ◽  
Future Directions ◽  
Diagnostic Pathology ◽  
The Past ◽  
Transmission Electron ◽  
Organ Systems ◽  
The Many

The role of electron microscopy and associated techniques is assured in diagnostic pathology. At the present time, most of the progress has been made on tissues examined by transmission electron microscopy (TEM) and correlated with light microscopy (LM) and by cytochemistry using both plastic and paraffin-embedded materials. As mentioned elsewhere in this symposium, this has revolutionized many fields of pathology including diagnostic, anatomic and clinical pathology. It began with the kidney; however, it has now been extended to most other organ systems and to tumor diagnosis in general. The results of the past few years tend to indicate the future directions and needs of this expanding field. Now, in addition to routine EM, pathologists have access to the many newly developed methods and instruments mentioned below which should aid considerably not only in diagnostic pathology but in investigative pathology as well.

Cryomicroscopy of multiphase latices

1991 ◽  
Vol 49 ◽  
pp. 1060-1061
Author(s):  
O. L. Shaffer ◽  
M.S. El-Aasser ◽  
C. L. Zhao ◽  
M. A. Winnik ◽  
R. R. Shivers
Keyword(s):  
Electron Microscopy ◽  
Radiation Damage ◽  
Freeze Fracture ◽  
Particle Morphology ◽  
Second Stage ◽  
Transmission Electron ◽  
Important Approach ◽  
Carbon Coated ◽  
Preparation Techniques

Transmission electron microscopy is an important approach to the characterization of the morphology of multiphase latices. Various sample preparation techniques have been applied to multiphase latices such as OsO4, RuO4 and CsOH stains to distinguish the polymer phases or domains. Radiation damage by an electron beam of latices imbedded in ice has also been used as a technique to study particle morphology. Further studies have been developed in the use of freeze-fracture and the effect of differential radiation damage at liquid nitrogen temperatures of the latex particles embedded in ice and not embedded.Two different series of two-stage latices were prepared with (1) a poly(methyl methacrylate) (PMMA) seed and poly(styrene) (PS) second stage; (2) a PS seed and PMMA second stage. Both series have varying amounts of second-stage monomer which was added to the seed latex semicontinuously. A drop of diluted latex was placed on a 200-mesh Formvar-carbon coated copper grid.

Transmission Electron Microscopy studies of texture of Cr underlayer of magnetic recording hard disk

1991 ◽  
Vol 49 ◽  
pp. 580-581
Author(s):  
L. Tang ◽  
G. Thomas ◽  
M. R. Khan ◽  
S. L. Duan
Keyword(s):  
Electron Microscopy ◽  
Thin Films ◽  
Transmission Electron Microscopy ◽  
Magnetic Recording ◽  
Magnetic Thin Films ◽  
Magnetic Films ◽  
Substrate Bias ◽  
Epitaxial Relationship ◽  
Transmission Electron

Cr thin films are often used as underlayers for Co alloy magnetic thin films, such as Co1, CoNi2, and CoNiCr3, for high density longitudinal magnetic recording. It is belived that the role of the Cr underlayer is to control the growth and texture of the Co alloy magnetic thin films, and, then, to increase the in plane coercivity of the films. Although many epitaxial relationship between the Cr underlayer and the magnetic films, such as ﹛1010﹜Co/ {110﹜Cr4, ﹛2110﹜Co/ ﹛001﹜Cr5, ﹛0002﹜Co/﹛110﹜Cr6, have been suggested and appear to be related to the Cr thickness, the texture of the Cr underlayer itself is still not understood very well. In this study, the texture of a 2000 Å thick Cr underlayer on Nip/Al substrate for thin films of (Co75Ni25)1-xTix dc-sputtered with - 200 V substrate bias is investigated by electron microscopy.

Transmission Electron Microscopy of Evaporated Perylene Thin Films

1990 ◽  
Vol 48 (2) ◽  
pp. 336-337
Author(s):  
C. Ewins ◽  
J.R. Fryer
Keyword(s):  
Electron Microscopy ◽  
Thin Films ◽  
Transmission Electron Microscopy ◽  
Molecular Electronics ◽  
High Vacuum ◽  
Amorphous Layer ◽  
Electric Heater ◽  
Transmission Electron ◽  
Polycyclic Aromatic

The preparation of thin films of organic molecules is currently receiving much attention because of the need to produce good quality thin films for molecular electronics. We have produced thin films of the polycyclic aromatic, perylene C10H12 by evaporation under high vacuum onto a potassium chloride (KCl) substrate. The role of substrate temperature in determining the morphology and crystallography of the films was then investigated by transmission electron microscopy (TEM).The substrate studied was the (001) face of a freshly cleaved crystal of KCl. The temperature of the KCl was controlled by an electric heater or a cold finger. The KCl was heated to 200°C under a vacuum of 10-6 torr and allowed to cool to the desired temperature. The perylene was then evaporated over a period of one minute from a molybdenum boat at a distance of 10cm from the KCl. The perylene thin film was then backed with an amorphous layer of carbon and floated onto copper microscope grids.

Fine structure and possible functions of the hermaphrodite duct of some pulmonate snails (Mollusca: Gastropoda)

1990 ◽  
Vol 48 (3) ◽  
pp. 68-69
Author(s):  
Alan N. Hodgson
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Seminal Vesicle ◽  
Reproductive Biology ◽  
Light Microscope ◽  
Light Microscope Level ◽  
Transmission Electron ◽  
Standard Techniques

The hermaphrodite duct of pulmonate snails connects the ovotestis to the fertilization pouch. The duct is typically divided into three zones; aproximal duct which leaves the ovotestis, the middle duct (seminal vesicle) and the distal ovotestis duct. The seminal vesicle forms the major portion of the duct and is thought to store sperm prior to copulation. In addition the duct may also play a role in sperm maturation and degredation. Although the structure of the seminal vesicle has been described for a number of snails at the light microscope level there appear to be only two descriptions of the ultrastructure of this tissue. Clearly if the role of the hermaphrodite duct in the reproductive biology of pulmonatesis to be understood, knowledge of its fine structure is required.Hermaphrodite ducts, both containing and lacking sperm, of species of the terrestrial pulmonate genera Sphincterochila, Levantina, and Helix and the marine pulmonate genus Siphonaria were prepared for transmission electron microscopy by standard techniques.

Magnetic and dielectric properties of BiFeO3 nanoparticles

2015 ◽  
Vol 7 (2) ◽  
pp. 1393-1403
Author(s):  
Dr R.P VIJAYALAKSHMI ◽  
N. Manjula ◽  
S. Ramu ◽  
Amaranatha Reddy
Keyword(s):  
Diffuse Reflectance Spectrum ◽  
Precipitation Method ◽  
Secondary Phases ◽  
X Ray Diffraction ◽  
Bifeo3 Nanoparticles ◽  
Transmission Electron ◽  
Multiferroic Bifeo3 ◽  
Pure Phase ◽  
Simple Chemical ◽  
Co Precipitation

Single crystalline nano-sized multiferroic BiFeO3 (BFO) powders were synthesized through simple chemical co-precipitation method using polyethylene glycol (PEG) as capping agent. We obtained pure phase BiFeO3 powder by controlling pHand calcination temperature. From X-ray diffraction studies the nanoparticles were unambiguously identified to have a rhombohedrally distorted perovskite structure belonging to the space group of R3c. No secondary phases were detected. It indicates single phase structure. EDX spectra indicated the appearance of three elements Bi, Fe, O in 1:1:3. From the UV-Vis diffuse reflectance spectrum, the absorption cut-off wavelength of the BFO sample is around 558nm corresponding to the energy band gap of 2.2 eV. The size (60-70 nm) and morphology of the nanoparticles have been analyzed using transmission electron microscopy (TEM).   Linear M−H behaviour and slight hysteresis at lower magnetic field is observed for BiFeO3 nanoparticles from Vibrating sample magnetometer studies. It indicates weak ferromagnetic behaviour at room temperature. From dielectric studies, the conductivity value is calculated from the relation s = L/RbA Sm-1 and it is around 7.2 x 10-9 S/m.

Effect of synthesis and activation methods on the character of CoMo/ultrastable Y-zeolite catalysts

2021 ◽  
Vol 19 (1) ◽  
pp. 745-754
Author(s):  
Khoirina Dwi Nugrahaningtyas ◽  
Eddy Heraldy ◽  
Rachmadani ◽  
Yuniawan Hidayat ◽  
Indriana Kartini
Keyword(s):  
Electron Microscopy ◽  
Reduction Process ◽  
Particle Morphology ◽  
Zeolite Catalysts ◽  
X Ray Diffraction ◽  
Y Zeolite ◽  
X Ray ◽  
Irregular Particle ◽  
Transmission Electron ◽  
Pure Metals

Abstract The properties of three types of CoMo/USY catalysts with different synthesized methods have been studied. The sequential and co-impregnation methods followed by activation using calcination and reduction process have been conducted. The properties of the catalysts were examined using Fourier-transform-infrared (FTIR) spectroscopy, X-ray diffraction (XRD) with refinement, and surface area analyzer (SAA). The FTIR spectrum study revealed the enhanced intensity of its Bronsted acid site, and the XRD diffractogram pattern verified the composition of pure metals, oxides, and alloys in the catalyst. The SAA demonstrated the mesoporous features of the catalyst. Scanning electron microscopy showed an irregular particle morphology. Additional analysis using the transmission electron microscopy indicated that the metal has successfully impregnated without damaging the USY structure.

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