Pathohistology

 

All tissue removed from people undergo histopathological examination in Hungary, which ensures that the laesions are given a proper histopathological diagnosis. This examination helps to identify the nature of the laesion, the extension, and the possibility of metastases.

 

Formalinban fixált, feldolgozásra váró minta

The sample, along with the clinician’s application form, usually comes fixated in formaline to the Institute of Pathology. Formaline fixation provides a long lasting preservation of the structure of the tissue. The sample is at first precisely identified by an assistant according to the data on the clinician’s application form.

 

 

 

 

 Excision

 

A patológus éles késsel, pengével kivágja a szövettani beágyazásra, mikroszkópos vizsgálatra kerülő területeket.During the process of the so called „excision”, the pathologist describes the major characteristics of the recogniseable tissue, the organ, or the parts of the organ. The pathologist also observes the colour, the surface, the structure of the sample, and then cuts certain parts with a sharp blade or knife for further microscopic evaluation. The size of the sample is standardized to reach optimal efficacy of the process.

The pathologist performing the excision registers the number of the tissue blocks and the cassettes, which are used to store the sample cut for further analysis. The pathologist also describes the status of the sample that remained, including the state of fixation, and the size of the tissue.

 

 

 

If it is needed, the pathologist may apply indian ink to the edge of the surgical excision, or ask for special staining methods, such as fat staining or decalcination. In this case, the assistant places the sample into a different cassette, marking the procedure that was indicated, along with the identification number of the histopathological case. The unfixated, fresh samples are stored in formaline for one day.

 

 

Paraffine saturation

 

To create a slide suitable for microscopic histopathological examination, a further, usually 24 hour long process is needed. First, embedding the tissue sample includes the removal of water, which is followed by paraffine saturation.

 

 

 

 

Automatic paraffine saturatin machine (left), manual paraffine saturation (middle), sample saturated in paraffine (right).

 

Creating a slide

 

 

Paraffine hardens the sample, making it suitable for cutting performed by a microtome.  Certain parts of the paraffine block is then cut into 2-5 µm thick slices, which are applied to a slide by a very delicate technique by an assistant.

 

 

 

 

 

For proper microscopic evaluation, certain staining methods are available, some of them standard, some of them combinated and rarely used. The routine method in pathology is the so called haematoxiline-eosine (HE) staining. If the diagnosis of the laesion is not possible with regular techniques, special staining procedures (Tsiebel or Groscott) are used.

To cover the slides that went through staining, covering is used by an extremely thin, tenth of a millimeter thick glass cover. This also preserves the condition and colour of the slide.

 

 

 

Automatic covering machine (left) an appliance for hematoxylin-eosine staining (right).

 

Microscopic evaluation

 

Mikroszkóp

During microscopic histopathological examination, the pathologist summarizes the data on  the clinician’s application form, views the previous clinical information available, and conceives the question taken by the clinician referreing to the nature of the laesion, along with the macroscopic description. The examination includes identification of the usual and unusual tissue, their description and their relation to each other. The pathologist may also select the tissue blocks that need further special staining, immunohistochemistry, or even do a new excision from a remaining tissue mentioned above.

 

 

 

 

In the next four slides, we would like to demonstrate beningant laesions (on the right), as well as malignant diseases (on the left). These photos are from the collection of Prof. Dr. Zoltán Nemes.

 

Hydradenoma vulvae (left) - benignant laesion and carcinoma planocellulare penis (right) - a malignant disease.. 

 

 

Prof. Dr. Nemes Zoltán gyűjteményéből: balra: Naevus naevocellularis dermo-epidermalis; jobbra: Melanoma malignum nodularis typus

 

 

 

 

 

 

 

Naevus naevocellularis dermo-epidermalis (left) - a benignant laesion and melanoma malignum nodular type  (right) - a malignant disease.

(The photos are a property of Prof Dr. Zoltán Nemes.)

 

 

 

Immunohistochemistry

 

Automata berendezés immunhisztokémiai vizsgálatokhoz

 If problems occur with differential diagnosis, immunohistochemical procedures are available to the pathologist to provide a most precise diagnosis. We demonstrate an S-100 positivity in case of a malignant melanoma along with the microscopic picture of the reaction.

 

 

 

 

 

                                                    

 S-100 positivity in case of a malignant melanoma.

 

 

 Histopathological report 

 

The result of the pathologist’s work is the histopathological report. This contains the basic data of the patient, clinical information, macroscopic and microscopic description of the laesion, the diagnosis, and may contain remarks or comments  on the differential-diagnosis, or suggestions on further therapy. In the Institute of Pathology, regular personal consultation is available for every clinician with the pathologist.  

After the report is handed out, the slides are stored according to the case number. The Institute of Pathology preserves each slide for 10 years.

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