scholarly journals Non-contact acoustic capture of microparticles from small plasma volumes

Lab on a Chip ◽  
2015 ◽  
Vol 15 (12) ◽  
pp. 2588-2596 ◽  
Author(s):  
Mikael Evander ◽  
Olof Gidlöf ◽  
Björn Olde ◽  
David Erlinge ◽  
Thomas Laurell
Keyword(s):  
Contact Method ◽  
Differential Centrifugation ◽  
Limited Volume ◽  
Centrifugation Method

An acoustic non-contact method for extracting platelet-derived microparticles from small plasma volumes is presented. The method has higher recovery and can handle smaller sample volumes than conventional differential centrifugation method. It is optimal for the analysis of precious biobank samples with limited volume.

Radioimmunoassay of human thyroglobulin with use of "thyroglobulin-free" plasma prepared by ultracentrifugation as diluent.

1980 ◽  
Vol 26 (11) ◽  
pp. 1566-1568 ◽  
Author(s):  
K Ikekubo ◽  
J Jutton ◽  
A B Schneider
Keyword(s):  
Total Thyroidectomy ◽  
Human Plasma ◽  
Normal Subjects ◽  
New Method ◽  
Differential Centrifugation ◽  
Thyroid Malignancy ◽  
Centrifugation Method ◽  
Preparation And Evaluation ◽  
Free Plasma

Abstract We describe the preparation and evaluation of a new diluent for use in preparing standard curves for the radioimmunoassay of human thyroglobulin (TG). This diluent, "TG-free human plasma, allows measurement of TG in lower concentrations than with previous assay systems because the components of the standards are more similar to those of the samples. TG-free plasma was prepared by a modification of the differential centrifugation method of Edelhoch (J. Biol. Chem. 235:1326, 1960). Standard curves prepared by use of TG-free plasma and plasma obtained from thyroidectomized patients were identical. In our assay, plasma from patients who had undergone total thyroidectomy produced no displacement when 25 to 100 microL of plasma was assayed. Twenty-five normal subjects all had detectable TG concentrations, ranging from 4.0 to 25.7 microgram/L. Only 27 of 2000 determinations performed by this new method had undetectable values and 26 of these were from patients with total or near-total thyroidectomies. TG concentration determined by this assay may be more sensitive for finding recurrent thyroid malignancy.

scholarly journals Selective loss of microvesicles is a major issue of the differential centrifugation isolation protocols

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Annamaria Nigro ◽  
Annamaria Finardi ◽  
Marzia M. Ferraro ◽  
Daniela E. Manno ◽  
Angelo Quattrini ◽  
...  
Keyword(s):  
Common Mechanism ◽  
Differential Centrifugation ◽  
Selective Loss ◽  
Pathological Conditions ◽  
Centrifugation Step ◽  
Centrifugation Method ◽  
Downstream Analysis ◽  
Cell Conditioned Medium ◽  
Rotor Type ◽  
Centrifugation Protocol

AbstractMicrovesicles (MVs) are large extracellular vesicles differing in size, cargo and composition that share a common mechanism of release from the cells through the direct outward budding of the plasma membrane. They are involved in a variety of physiological and pathological conditions and represent promising biomarkers for diseases. MV heterogeneity together with the lack of specific markers had strongly hampered the development of effective methods for MV isolation and differential centrifugation remains the most used method to purify MVs. In this study, we analysed the capacity of the differential centrifugation method to isolate MVs from cell-conditioned medium using flow cytometry and TEM/AFM microscopy. We found that the loss of MVs (general population and/or specific subpopulations) represents a major and underestimate drawback of the differential centrifugation protocol. We demonstrate that the choice of the appropriate rotor type (fixed-angle vs swinging-bucket) and the implementation of an additional washing procedure to the first low-speed centrifugation step of the protocol allow to overcome this problem increasing the total amount of isolated vesicles and avoiding the selective loss of MV subpopulations. These parameters/procedures should be routinely employed into optimized differential centrifugation protocols to ensure isolation of the high-quantity/quality MVs for the downstream analysis/applications.

Peroxidase Localization in Hartmanella Trophozoites

1971 ◽  
Vol 29 ◽  
pp. 346-347 ◽  
Author(s):  
George E. Childs ◽  
Joseph H. Miller
Keyword(s):  
Hydrogen Peroxide ◽  
Ultrastructural Localization ◽  
Pathogenic Strain ◽  
Oxidative Enzymes ◽  
Differential Centrifugation ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
The Subject ◽  
Axenic Cultures

Biochemical and differential centrifugation studies have demonstrated that the oxidative enzymes of Acanthamoeba sp. are localized in mitochondria and peroxisomes (microbodies). Although hartmanellid amoebae have been the subject of several electron microscopic studies, peroxisomes have not been described from these organisms or other protozoa. Cytochemical tests employing diaminobenzidine-tetra HCl (DAB) and hydrogen peroxide were used for the ultrastructural localization of peroxidases of trophozoites of Hartmanella sp. (A-l, Culbertson), a pathogenic strain grown in axenic cultures of trypticase soy broth.

3-D organization of fibrinogen receptors using computer reconstruction and whole-mount microscopy

1988 ◽  
Vol 46 ◽  
pp. 30-31
Author(s):  
E. T. O'Toole ◽  
R. R. Hantgan ◽  
J. C. Lewis
Keyword(s):  
Whole Blood ◽  
Colloidal Gold ◽  
Blood Platelets ◽  
Cell Contact ◽  
Computer Assisted ◽  
Adherent Cells ◽  
Differential Centrifugation ◽  
Computer Reconstruction ◽  
Sds Page ◽  
Whole Mount

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.

Ultrastructural Characteristics of Platelets Separated from Plasma by ADP-Induced Aggregation

1976 ◽  
Vol 34 ◽  
pp. 164-165
Author(s):  
B.A. Shinoda ◽  
M.D. Hardison ◽  
S.F. Mohammad ◽  
H.Y.K. Chuang ◽  
R.G. Mason
Keyword(s):  
Blood Platelets ◽  
Gel Filtration ◽  
Differential Centrifugation ◽  
Ultrastructural Characteristics ◽  
Induced Aggregation

The utilization of blood platelets in experimentation frequently requires their separation from blood and subsequent resuspension in media of known composition. Several methods are available for preparation of isolated platelets (1-3) by differential centrifugation or gel filtration, but most methods are tedious and time consuming. Often platelets obtained by use of such methods are in a state different functionally and ultrastructurally from that of platelets in plasma (4).Recently Mohammad, Reddick, and Mason (5) reported a method in which platelets were separated from plasma by ADP-induced aggregation, washed several times, and then incubated in a carefully selected medium that resulted in deaggregation of platelets.

Pathophysiological function of the mitochondria as a calcium reservoir: Quantitative x-ray microanalysis

1990 ◽  
Vol 48 (2) ◽  
pp. 164-165
Author(s):  
K. Teraoka ◽  
N. Kaneko ◽  
Y. Horikawa ◽  
T. Uchida ◽  
R. Matsuda ◽  
...  
Keyword(s):  
Bolus Injection ◽  
Contact Method ◽  
X Ray

The aim of this study was to elucidate the role of the mitochondria as a store of calcium(Ca) under the condition of pathophysiological Ca overload induced by a rise in extracellular Ca concentration and the administration of isoproterenol.Eight rats were employed, and hearts were perfused as in the Langendorff method with Krebs-Henseleit solution gassed with 95% O2 and 5% CO2. Tow specimens were perfused with 2mM Ca for 30 min, and 2 were perfused with 5.5 mM Ca for 20 min. 4 specimens were perfused with 2 mM Ca for 5 min, and of these 4, 2 were infused with 10-7 mM/kg/min. isoproterenol for 5 min, and 2 were given a bolus injection of 3 x 10-7 mM isoproterenol. After rapid-cryofixation by the metal-mirror contact method with a Reichert-Jung KF80/MM80, and cryosectioning at -160 to -180° C with a Reichert-Jung Ultracut Fc-4E, ultrathin specimens (100nm) were free-ze-dreid for several hours at 10-5 Torr in the JEOL FD 7000, and mitochondrial Ca was determined by quantitative x-ray micranalysis (JEOL 1200EX, LINK AN 10000S).

Early ossicle growth in the regenerating disc of a brittle star

1994 ◽  
Vol 52 ◽  
pp. 364-365
Author(s):  
M. Shlepr ◽  
R. L. Turner
Keyword(s):  
Cell Membrane ◽  
Light Microscopy ◽  
De Novo ◽  
Current Model ◽  
Syncytium Formation ◽  
Brittle Star ◽  
Intracellular Location ◽  
Limited Volume ◽  
Tissue Calcification

Calcification in the echinoderms occurs within a limited-volume cavity enclosed by cytoplasmic extensions of the mineral depositing cells, the sclerocytes. The current model of this process maintains that the sheath formed from these cytoplasmic extensions is syncytial. Prior studies indicate that syncytium formation might be dependent on sclerocyte density and not required for calcification. This model further envisions that ossicles formed de novo nucleate and grow intracellularly until the ossicle effectively outgrows the vacuole. Continued ossicle growth occurs within the sheath but external to the cell membrane. The initial intracellular location has been confirmed only for elements of the echinoid tooth.The regenerating aboral disc integument of ophiophragmus filograneus was used to test the current echinoderm calcification model. This tissue is free of calcite fragments, thus avoiding questions of cellular engulfment, and ossicles are formed de novo. The tissue calcification pattern was followed by light microscopy in both living and fixed preparations.

Sign in / Sign up

Export Citation Format

Share Document