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    Home > Biochemistry News > Biotechnology News > The determination of multi-dyed red blood cell microkernels in mice bone marrow.

    The determination of multi-dyed red blood cell microkernels in mice bone marrow.

    • Last Update: 2020-10-30
    • Source: Internet
    • Author: User
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    . 1.
    Experimental principle The whole chromosome without silky fragments of chromosomes or chromosomes, or lost due to damage to spindle bodies, is still left in the cytoste in the later stages of cell division. After the end of the period, one or more rules of the secondary nucleus is formed separately, is contained in the cytonal of the subcell, because it is smaller than the main nucleus, so called the micro nucleus. The size should be 1/20 to
    1/20
    the
    core of the
    . The recactive rate and cytochemical reaction properties of the micro-core are the same as among the main nucleations. Microkernel rate is positively related to the dose or radiation accumulation effect of the drug, so simple, fast and sensitive micro-core counting can be used instead of complex distortion chromosome counts.
    Matter
    and
    Schmid
    established micro-core testing is an ideal method, at home and abroad have been used in radiation damage, chemical mutagents, new drug tests, chromosomal genetic diseases and pre-cancer diagnosis and other aspects.Poly-dyed red blood cells are a stage in the development of red blood cells from an early age to mature red blood cells, at which point the main nucleus of red blood cells has been excreted, because the cytoste contains cytosomes,
    Giemsa
    stained gray-blue, mature red blood cells have disappeared, dyed pale orange. The number of red blood cells in the bone marrow is sufficient, and because it is nuclear-free, it is easy to observe micro-cores, so poly-dyed red blood cells in the bone marrow become the preferred cell group for micro-nuclear testing.. 2.
    subjects Adult healthy mice, weighing
    25g
    , both male and female.. 3.

    Reagents
    and Equipment(
    1
    ) Equipment:
    Microscope
    , Low Speed
    Centrifuge
    , Electronics
    Balance
    Anatomy devices (enamel anatomical discs, probes, scissors, tweezers), syringes,
    tubular tubes
    , test tube racks, abrasive reagent bottles, droppers, dyeing cylinders, ear wash balls,
    -measure barrels
    , cleaning slides.(
    2
    ) reagents:
    0.9
    % physiological saline, inactivate calf
    serum
    , cyclophosphamide, methanol,
    PBS (pH6.8)
    ,
    Giemsa
    dye, Rhys dye.. 4.
    Experimental method and step(
    1
    ) cyclophosphamide treatment:
    24h
    before taking bone marrow, the mouse abdominal cavity was injected with cyclophosphamide at a dose of
    100mg
    /
    kg
    animal body weight.(
    2
    ) Take bone marrow: Kill the mice with spinal cord injury, then use scissors to cut the skin and muscles off the thighs, remove the thigh bones, and use a small piece of gauze back and forth to clean the muscle residue attached to the bones. Cut off the puffed joint heads at both ends of the syringe, then use a syringe to absorb
    5 ml
    of physiological saline, insert it into one end of the collarbone, and rinse the bone marrow cells into a
    10 ml
    centrifuge tube. Rinse repeatedly until the bone marrow cavity is white.(
    3
    ) centrifugation treatment: the resulting cell suspension at
    1000rpm
    centrifugation
    10min
    , suck up the liquid, add
    2
    drops of inactivated calf serum to the sediment, make cell suspension.(
    4
    ) smear: drop a drop of suspension at one end of the slide, smear according to the conventional method, dry in the air.(
    5
    ) fixation: Put the dried slides in a methanol fixation liquid
    10min
    and remove to dry.(
    6
    ) staining: flat on the glass plate, with
    1
    :
    9
    :
    10


    :

    PBS
    : Rui's dye, dyeing
    15min
    , after washing dry mirror.(
    7
    ) observation: poly-dyed red blood cells are young red blood cells, gray-blue, slightly larger than red blood cells (red), micro-cores are round and oval, blue-purple or fuchsia (Figure
    13-3
    ).. (
    8
    ) Results Analysis: Under a microscope, each mouse bone marrow smear specimen count
    1000 to 2000
    poly-infected red blood cells, including the number of cells in which the micro nucleus appears, fills in the table, and calculates the microcycle rate, observing the record results (model
    13-2
    ).was injected with cyclophosphamide in the abdominal cavity of mice at a
    100 mg/kg
    body weight dose, and their poly-infected red blood cell microkernel rate (
    48.9
    ±
    3.6
    ).) Normal mice with poly-infected red blood cell microkernels had a microkernel rate of
    5
    per 1,000, and more than
    5
    per 1,000 were abnormal.
    . Question 1 1
    . What are the four main points of the conventional outer blood staining system? 2
    . What causes the chromosomes under the mirror to be too short, not dispersed, or the chromosomes to be lost? 3
    . What is the role of PHA
    , autumn daffodils and

    0.4% KCl
    solutions in the process of
    blood culture and dyeing system tablets? 4
    . What are the methods of human chromosome research? What are the characteristics of each? 5
    . What types of chromosomes are visible? . (Shu Qing) .
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