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    Home > Biochemistry News > Biotechnology News > Differential centrifugation separates the nucle nucleations of plant (animal) cells.

    Differential centrifugation separates the nucle nucleations of plant (animal) cells.

    • Last Update: 2020-10-20
    • Source: Internet
    • Author: User
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    Preparation and observation of mitochondrials and nucleosomes
    using the difference in the rate of subsidion between the nucleosomes and mitochondrials in a certain medium, the cell nucleosomes and mitochondrials can be separated step by step by step by the method of differential centrifugation. (Differential centrifugation technology)
    mitochondrials are important cells unique to the uclear cells for energy conversion. The animal and plant
    tissue
    into a homogenous slurry, in the appropriate suspension medium differential centrifugation method can separate the cell mitochondrials. In a certain centrifuge field (selecting a certain speed of
    centrifuges
    ), the velocity of the glocular particles depends on their density, radius and viscosity of the suspended medium. Centrifugation in a uniform suspended medium for a certain period of time, the various cytocytes and other inclusions in the tissue homogenizer will stay in different positions due to the different sembation velocity. By increasing centrifugal forces and centrifugal time in turn, these particles can be collected in batches by sequentizing them at the bottom of
    centrifugal tubes
    in batches of their size and weight. The sequence of cell sequential sequential is the nucleation, mitochondrial, lysosome and other microsomes, ucoseomes and macromolecules.
    suspension medium usually uses buffered sucrose solution, which is closer to the dispersion phase of cytoste, to a certain extent, can maintain the structure of the cytost and enzyme activity; The sample should be kept at 0 to 4 degrees C throughout the operation process to avoid enzyme inferation.The determination of
    organocyte marker enzyme is the main basis for evaluating the endometrial components and separation purity of the organe, such as the distribution of cytochrome
    oxidase
    on the membrane of the mitochondrial body, which keeps janus green B dye blue-green in the oxidation state, so that the mitochondrial color, and the dye in the cytoplocyte is reduced to colorless.
    Jenners Green B is a living dye that non-toxic staining of the cells or tissues of plants and animals in a live state. Because the surface of the dye (alkaline dye) with cations, acidic dyes with anions on the surface of the gel grain, and the dye itself has cations or anions, so that they attract each other, the dye is stacked down. Staining can reveal the authenticity of the existence of a natural structure in living cells without affecting the life activity of cells and producing any physical or chemical changes that can lead to cell death.
    Gimesa is a compound dye, i.e. a combination of acidic dyes and alkaline dyes. Ionized in an aqueous solution is a dye ion with positive and negative electricity.
    : Rat liver, pig liver
    s
    reagents
    :
    (1) 0.25mol/L sucrose-0.01mol/L trihydroxymethymeethane (Tris)-hydrochloric acid buffer (pH7.4)
    0.1mol/L trihydroxymethymethyl methane solution Liquid 10ml
    0.1mol/L hydrochloric acid 8.4ml
    plus double steamed water to 100ml, plus sucrose to make the concentration of 0.2mol/L
    (2) 0.34mol/L sucrose-0.01mol/LTris-hydrochloric acid buffer (pH7.4), with the same method.
    (3) 1% Janus green B (Janus green dye, said to take 50mg (pH7.4), Janus green B dissolved in 5 ml of physiological salt water, slightly
    heated
    to dissolve,
    filtered
    , that is, 1% original liquid.
    (4) Giemsa dye (Giemsa): called Giemsa powder 0.5g, glyceroid 33 ml, pure methanol 33ml. First add a small amount of glycerol to the Gimsa powder, then grind in the research until grain-free, and then pour the remaining glyceroid into the mix, 56 degrees C around 2h insulation to make it fully dissolved, and finally mixed with methanol, become the Kimsa original liquid, stored in a brown bottle. When used, a small amount is sucked out and diluted 10 to 20 times with 1/5mol/L phosphoric acid buffer.
    (5) 1/15 mol/L phosphoric acid buffer?(pH6.8)
    1/15 mol/L phosphate potassium dihydrohydrogen (KH2PO4)50 ml
    1/15 mol/L phosphate di sodium (Na2HPO4) 50 ml
    Cornay fixation: waterless ethanol 6 ml ice acetic acid 1 ml chloroform 3 ml
    .
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