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Temperaturpåverkan på lymfocyters stabilitet i helblod taget i TransFix/EDTA-rör
Jönköping University, School of Health and Welfare, HHJ, Dep. of Natural Science and Biomedicine.
Jönköping University, School of Health and Welfare, HHJ, Dep. of Natural Science and Biomedicine.
2019 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
The effect of temperature on lymphocyte subset stability in whole blood collected in TransFix/EDTA-tube (English)
Abstract [sv]

Vid flödescytometrisk analys av lymfocytpopulationer i helblod används EDTA-rör vilket kräver att analysen utförs inom 48 timmar, då cellens nedbrytningsprocess startar efter provtagning. Den korta hållbarheten för helblod medför även problem för interna kontroller som dessutom är kostsamma. Syftet med studien var att utvärdera om fryst, upptinat prov taget i TransFix/EDTA vakuumrör kan användas för analys av lymfocytpopulationer och lymfocytantigen, med flödescytometri. Syftet var även att utvärdera vilken koncentration av rören som är lämpligast att använda. Blodprover insamlade i två olika rör med spädning 1:5 och 1:10 från tolv blodgivare förvarades i -80°C.  Proverna tinades sedan upp i omgångar totalt tio gånger under sex veckor och analyserades med flödescytometri. Resultaten jämfördes med det första analyseringstillfället som utfördes på rumstempererade prover inom sex timmar efter provtagning. Resultaten visade att lymfocytkoncentrationen minskade i blodprover som förvarades i -80°C medan fördelningen av lymfocytpopulationerna hölls stabila. Det framgick i studien att TransFix/EDTA-röret med den lägre koncentrationen 1:10 av cellkonserveringsmedel ger bättre resultat vid förvaring i -80°C.

Abstract [en]

Flow cytometric analysis of lymphocyte subset is performed on whole blood, collected in EDTA vacutainer, requires analysis to be performed within 48 h, to prevent cellular degradation and to achieve reliable results. The short preservation of whole blood in EDTA vacutainer makes them not suitable as a laboratory quality control, because they are expensive. The purpose of this study was to evaluate if frozen and thawed blood samples, collected in TransFix/EDTA, can be analysed with flow cytometry without affecting the lymphocyte profile. The purpose of this study was also to evaluate which concentration of TransFix/EDTA vacutainer tube is most suitable for analysis. Blood samples were collected in two vacutainer tubes, concentration 1:5 and 1:10, from twelve blood donors and then stored in -80°C. The samples were then thawed ten times during a six-week period and analysed with flow cytometry. The result was compared to the result from the first analysis on room temperature samples that was performed within 6 h of veinpuncture collection. Some differences were observed in the concentration of CD45+ lymphocytes, but the percentage of lymphocyte subpopulations maintained stable in blood samples that was stored in -80°C when analysed with flow cytometry. The study showed that TransFix/EDTA vaccutainer tube with the low concentration 1:10 of cell preservative solutions shows better results on samples stored in -80°C.

Place, publisher, year, edition, pages
2019. , p. 20
Keywords [en]
Flow cytometry, TBNK panel, Freezing, Lymphocytes
Keywords [sv]
Flödescytometri, ToBNK-analys, Frysning, Lymfocyter
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:hj:diva-45237ISRN: JU-HHJ-BLA-1-20190091OAI: oai:DiVA.org:hj-45237DiVA, id: diva2:1333805
Subject / course
HHJ, Biomedical Laboratory Science
Supervisors
Examiners
Available from: 2019-07-04 Created: 2019-07-02 Last updated: 2019-07-04Bibliographically approved

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Värme, CarolineWirström, Johanna
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