PROCESSING OF TISSUE

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    drmithiladrmithila
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    Registered On: 14/05/2011
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    Depending on the processing procedure involved, the
    sample can be analyzed frozen, or embedded in paraffin
    or methacrylate, and can be examined under the electron
    microscope, or as a fresh sample. Molecular analyses are
    also possible.
    As commented above, frozen biopsies allow a rapid histopathological diagnosis. This procedure is usually used
    during surgery in order to allow immediate treatment
    decisions to be taken. However, the technique is contraindicated in the case of very hard tissues (bone or calcified
    tissue), lesions requiring more extensive study due to their
    complexity, or when no immediate diagnostic decision is
    required (3,6,7,12).
    Embedding in paraffin is indicated for light microscopic
    evaluation, and is the most commonly used technique.
    The sampled tissue is fixed in 10% formaldehyde solution.
    The most characteristic zones are selected from among the
    excess tissue resected. The specimen is processed by means
    of an automatized, serial hot paraffin embedding system
    involving progressive dehydration and final cooling and
    solidification, before sectioning with the microtome. The
    embedding process takes about 24 hours. Microtome sectioning is then carried out, mounting the specimens on slides
    and posteriorly removing the paraffin, with rehydration and
    staining as required. Finally, the specimen is again dehydrated and sealed with a coverslip. The staining techniques
    used depend on the type of histological study made (1).
    On the other hand, embedding in methacrylate is used for
    electron microscopic studies, and when hard tissues or materials are involved. Electron microscopy is able to reveal
    a series of morphological features that distinguish normal
    cells from tumor cells. It is also able to establish the extent
    of the tumor (12).
    Fresh specimen studies require transport of the biopsy
    sample as quickly as possible to the laboratory, for immunofluorescence evaluations. The specimens are to be
    humidified with saline solution to prevent drying. Before
    and during transport, the tissue can be kept refrigerated
    (2-8ºC), though freezing must be avoided (17).
    Immunofluorescence techniques in turn are either direct or
    indirect, depending on whether the test antigen is detected
    with a first labeled antibody or using a second anti-antibody,
    respectively (18).
    In our setting, these procedures are useful for diagnosing
    mucocutaneous diseases of an autoimmune nature, such as
    pemphigus, pemphigoid, erythema multiforme and lichen
    planus (Table 1).
    In the case of molecular studies, genetic molecular markers
    allow us to detect alterations before changes in cell morphology occur or clinical manifestations develop. Analysis of
    such molecular alterations is objective, and aims to identify
    specific genetic anomalies.E508
    Med Oral Patol Oral Cir Bucal. 2007 Nov 1;12(7):E504-10. Oral biopsy
    VULGAR PEMPHIGUS PEMPHIGOID
    ERYTHEMA
    MULTIFORME
    LICHEN PLANUS
    DIF
    IgG 100% C3 40% IgM,
    Fibrinogen
    IgG (particularly)
    C3 IgM IgA negative
    Fibrinogen (particularly)
    IgG 35%
    IgM 30%
    IIF Ab IgG 80-90% negative negative negative
    LUPUS
    ERYTHEMATOSUS
    LINEAL IgA
    DERMATOSIS
    HERPETIFORM
    DERMATITIS
    EPIDERMOLYSIS
    BULLOSA
    DIF
    IgG IgA IgM C3 C4
    IgA
    IgA
    C3
    IgG
    C3
    IIF negative negative negative negative
    table 1. Evaluation of test positivity for diagnosis via direct / indirect immunofluorescence in application to autoimmune
    mucocutaneous diseases.
    DIF: Direct immunofluorescence; IIF: Indirect immunofluorescence; Ig: Immunoglobulin; C: Complement fraction
    The principal oral cavity carcinogens are chemical (tobacco
    smoke), physical (radiation) and infectious agents (papillomavirus, Candida); as mutagens, they are able to induce
    structural changes in genes and chromosomes through point
    mutations, deletions, insertion or rearrangements. However,
    some changes of this kind can also occur spontaneously.
    Such genetic alterations taking place in the context of
    carcinogenesis can be used as targets for the detection of
    tumor cells in clinical samples (6).
    6) Purpose of the biopsy:
    Biopsies can be performed for diagnostic or experimental
    purposes. Thus, apart from diagnosis with a view to treatment, a biopsy can yield important information for the
    histopathological study of new disease entities. With this
    aim in mind, studies are made in laboratory animals, and
    exceptionally also in humans, with the obtainment of samples that can yield valuable information on the evolution of
    a given disease process

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