How to Diagnose Tumors and Infectious Organisms?

By: Tayyaba Bhatti

How to Diagnose Tumors and Infectious Organisms?

When people go through any minor or a major surgery their consultant advises histopathology of the removed tissue. Mostly patients don’t know what histopathology is, but they still submit their biopsy sample in a lab for histopathology.

Histopathology is the study of diseased tissue that is obtained through a biopsy by a pathologist. Tissue samples of the tissue are studied under a microscope in order to identify how a disease develops. The tissue sample goes through a special process so it can be analyzed on a slide. Microscopically, final diagnosis is made and reports are being issued to the patient.
But in few cases reports are delayed due to some reasons such as in case of requirement of previous surgery, radiological findings and any other history, whereas some reports are delayed as some special test are advised. For example,
Immunochemistry is the identification of a certain antigen in a histological tissue section or cytological preparation by an antibody specific to that antigen. Immunohistochemistry refers specifically to histological tissue sections.

  • It is extremely important research technique.

  • Antibodies can be raised against almost any type of antigen (protein, carbohydrate, lipid...) bind to antigen in a specific manner can be detected in several ways:

    1. a)Through fluorescent substances (fluorophores) – “Immunofluorescence technique”.

      b)Through enzymatic conversion (often by horseradish peroxidase, HRP) of a soluble substrate (chromogen) into a non-soluble and colorful reaction product “Immunoenzyme technique”

In pathology labs, IHC is used to diagnose a cancer as benign or malignant, determine the stage and grade of a tumor, and identify the cell type and origin of a metastasis to find the site of the primary tumor.

In IHC antibodies (matching molecules) are used that can seek out, identify and attach themselves to the markers on cells. The antibodies themselves can be seen under the microscope which helps the pathologist to make a precise identification.

Immunohistochemistry Steps:

First of all tissue section is deparffinize and then antigen is retrieve because in fixation process tissue proteins cross linked and ag-ab reaction is not possible. After retrieval endogenous peroxidase is blocked to prevent the background staining and tissue section is incubated with primary antibody if respective antigen is present in tissue then antigen-antibody complex will be formed. After that when tissue is incubated with secondary antibody it will get attached it self with primary antibody and a complex of ag-primary ab-secondary ab will be formed and produce color when substrate chromogen is added. These tissues are counterstained with hematoxylin. Dehydrate and cover with cover slip.

A positive control should always run along with the test tissue for the accurate interpretation of the results and also helps to check the efficiency of the reagent.

Finally, to diagnose tumors of any kind and infectious diseases, immunohistochemical stains are the best and advanced solution in diagnostic science. Chughtais Lahore Lab is the one of the leading diagnostic laboratories providing services for immunohistochemistry, biopsies, Frozen Sections, immunostains to Hospitals and Clinics all over the Pakistan. CLL have highly qualified pathologist, doctors in all departments along with Histopathology.

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Author:Tayyaba Bhatti is a professional histopathology technician at Chughtais Lahore Lab,
Diagnostic Laboratory | Immunohistochemistry

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