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Har hantering av objektglas vid snittning betydelse för kombinerad immunhistokemi & in situ hybridisering-färgning?
Malmö University, Faculty of Health and Society (HS).
2022 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
Does handling of sections during sectioning have significance for combined immunhistochemistry & in situ hybridization-staining? (English)
Abstract [sv]

Tillväxtfaktorreceptorn human epidermal growth factor receptor 2 (her2) är ett protein på ytan av cancerceller som stimulerar tillväxt, differentiering samt överlevnad av tumörceller. Överuttryck av her2-proteinet eller ett ökat antal kopior av her2-genen i tumörcellerna klassificeras som her2-positiva. Her2-positiv bröstcancer överensstämmer med en aggressiv form av bröstcancer och därmed med en försämrad prognos. Cirka 20 % av alla bröstcancerpatienter drabbas av her2-positiv bröstcancer. Bedömning av her2-status används som behandlingsprediktiv faktor för att stödja effekten av målstyrd antikroppsterapi med Trastuzumab i kombination med cytostatika. Identifiering av her2-genproteinuttryck utförs genom en kombinerad immunhistokemi (IHC) & in situ hybridiering (ISH) analys. Ca 20–25% av alla her2-genprotein analyser på klinisk patologi i Helsingborg, genomgår en omkörning på grund av ”bakgrundsdamm”, svaga eller ospecifika signaler. Syftet med studien var att undersöka om preanalytiska faktorer vid snittning påverkar resultatet vid en kombinerad immunhistokemi och in situ hybridisering-färgning. Befintligt formalinfixerad, paraffininbäddad bröstcancervävnad med känt her2-uttryck användes som provmaterial. Två sorters objektglas (FLEX IHC Microscope Slides, DAKO och Superfrost Ultra Plus, Thermo Scientific) och 2 inkubationstider vid bakning (1 timme och 2 timmar) samt nysnittad och 2 dygn kylskåpsförvarad vävnadssnitt testades i studien. Resultatet visade att det föreligger en skillnad mellan val av objektglas, samt nysnittad/kylskåpsförvarade vävnadssnitt, då studien visade att SuperFrost Ultra Plus på nysnittat vävnad ger bättre IHC & ISH-färgning. En förbättrad kombinerad IHC & ISH-färgning leder till säkrare bedömning om antikroppsbehandling med Tastuzumab (herceptin) är aktuell för patienten.

Abstract [en]

The human epidermal growth factor receptor 2 (her2) is a protein on the surface of cancer cells that stimulates the growth, differentiation and survival of tumour cells. Overexpression of the her2 protein or an increased number of copies of the her2 gene in the tumour cells are classified as her2 positive. Her2-positive breast cancer is consistent with an aggressive form of breast cancer and therefore with a poor prognosis. About 20% of all breast cancer patients suffer from her2-positive breast cancer. Assessment of her2 status serves as a treatment predictive factor to support the use of targeted antibody therapy with Trastuzumab in combination with cytostatic drugs. Identification of her2 gene protein expression performed by a combined immunohistochemistry (IHC) & in situ hybridization (ISH) assay. Approximately 20-25% of all her2 gene protein analyses in clinical pathology in Helsingborg undergo and overturning due to "background dust", weak or non-specific signals. The purpose of the study was to investigate preanalytical factors in workplace incisions at immunohistochemistry and in situ hybridization staining. Existing formalin-fixed, paraffin-embedded breast cancer tissue with known her2 expression was used as test material. Two types of slides (FLEX IHC Microscope Slides, DAKO and Superfrost Ultra Plus, Thermo Scientific) and 2 incubation times during baking (1 hour and 2 hours) as well as news and 2 days of refrigerated tissue sections were tested in the study. The results showed that there is a difference between the selection of slides, as well as newly cut / refrigerated tissue sections, as the study showed that SuperFrost Ultra Plus and newly cut tissue give better IHC & ISH staining. An improved combined IHC & ISH staining leads to safer assessment of antibody therapy with Tastuzumab (herceptin) is relevant for the patient.

Place, publisher, year, edition, pages
2022. , p. 22
Keywords [en]
Ductal invasive breast cancer, FLEX IHC Microscope Slides Dako, human epidermal receptor 2 immunohistochemistry, in situ hybridization, Superfrost Ultra Plus
Keywords [sv]
Duktal invasiv bröstcancer, FLEX IHC Microscope Slides Dako, immunhistokemi, in situ hybridisering, human epidermal receptor 2, Superfrost Ultra Plus
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:mau:diva-51354OAI: oai:DiVA.org:mau-51354DiVA, id: diva2:1656832
Educational program
HS Biomedicinsk laboratorievetenskap
Supervisors
Examiners
Available from: 2022-05-09 Created: 2022-05-08 Last updated: 2022-05-09Bibliographically approved

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