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Samenvatting Hematologie (labdiagnostiek en automatisatie) - MLT3 - HoGent
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Samenvatting van de ppt van labdiagnostiek en automatisatie hematologie 2
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Samenvatting Hematologie 2Inhoud
1. INLEIDING............................................................................................................................... 4
2. PRE-ANALYTISCHE FASE.......................................................................................................... 4
Bloedafname........................................................................................................................... 4
Onderscheid tussen plasma en serum.................................................................................... 4
Centrifugeren van bloed.......................................................................................................... 5
Chemische anticoagulantia..................................................................................................... 5
EDTA..................................................................................................................................... 5
Citraat..................................................................................................................................... 5
Heparine.............................................................................................................................. 5
Hemolysaat............................................................................................................................. 6
Factoren die bloeduitslag kunnen beïnvloeden....................................................................6
Bloedafnamefactoren........................................................................................................... 6
Bewaarcondities................................................................................................................... 6
Integriteit van staal.............................................................................................................. 7
3. HEMATO-MORFOLOGIE........................................................................................................... 7
Fixeren..................................................................................................................................... 7
Kleuren met May-Grunwald – Giemsa methode......................................................................7
Beoordelen onder microscoop................................................................................................. 8
Morfometrische elektronische differentiële telling..................................................................8
4. HEMOCYTOMETRIE................................................................................................................. 9
4.1. Celtellingen: microscopie................................................................................................. 9
4.1. Celtellingen: elektronisch................................................................................................. 9
Impedantie........................................................................................................................... 9
Optische methode met flow cytometrie............................................................................. 10
Hemocytometrie analyzer................................................................................................. 11
4.1.1 Tellen van erytrocyten............................................................................................... 12
4.1.2 Tellen van reticulocyten............................................................................................ 12
4.1.3. Elektronische telling van leukocyten........................................................................13
4.2. Hemoglobine.................................................................................................................. 17
Bepaling hemoglobineconcentratie....................................................................................17
4.3. Hematocriet................................................................................................................... 18
Hematocrietbepaling.......................................................................................................... 18
4.4. Erytrocytindices.............................................................................................................. 19
4.4.1 MCV (mean cell volume)........................................................................................... 19
4.4.2 MCH (mean cell hemoglobine).................................................................................. 19
4.4.3 MCHC (mean cell hemoglobine conc)........................................................................19
4.4.4 Erytrocytenvolumedistributie en RDW......................................................................19
1
, 4.5 Bezinkingssnelheid van erytrocyten (BSE)......................................................................20
4.5.1 Bezinking volgens Westergen................................................................................... 20
4.5.2. BSE........................................................................................................................... 22
5. HEMOSTASE.......................................................................................................................... 24
5.1. Pre-analytische fase....................................................................................................... 24
6. LABDIAGNOSTIEK PRIMAIRE HEMOSTASE.............................................................................25
6.1 LABOratoriumanalyses van de primaire hemostase........................................................25
Trombocytentelling............................................................................................................. 25
Bloedingstijd...................................................................................................................... 26
Platelet function analyser................................................................................................... 26
Aggregatieonderzoek......................................................................................................... 27
Flowcytometrie.................................................................................................................. 28
Secretieonderzoek............................................................................................................. 28
6.2 Labdiagnostiek van secundaire hemostase.....................................................................28
6.2.1 Inleiding.................................................................................................................... 28
6.2.2 Technieken................................................................................................................ 29
6.2.3 aPTT.......................................................................................................................... 29
6.2.4 PT.............................................................................................................................. 29
6.2.5 Dosage intrinsieke factoren...................................................................................... 30
6.2.6 Dosage extrinsieke factoren...................................................................................... 31
6.2.7 Trombinetijd.............................................................................................................. 31
6.2.8 Reptilasetijd (niet echt te kennen)............................................................................ 31
6.2.9 Fibrinogeenbepaling - Methode von Clauss...............................................................31
6.2.10 Clot solubility test................................................................................................... 33
6.2.11 Antitrombine........................................................................................................... 33
6.2.12 Proteïne C en S bepaling......................................................................................... 33
6.2.13. D-dimeertest.......................................................................................................... 33
7. PATHOLOGIE-HEMOSTASE BLOEDINGSNEIGINGEN................................................................34
7.1. Oorzaken bloedingsneigingen........................................................................................ 34
7.1.1. Stoornissen in de primaire hemostase.....................................................................34
7.1.2. Stoornissen in secundaire hemostase......................................................................40
8. IMMUUNHEMATOLOGIE......................................................................................................... 42
8.1 Bloedgroepantigenen...................................................................................................... 42
8.1.1 ABO-systeem............................................................................................................ 42
8.1.2. Lewis-systeem.......................................................................................................... 45
8.1.3. Rhesus-systeem....................................................................................................... 45
9. BLOEDSTOLLING - TROMBOFILIE.......................................................................................... 47
9.1. Trombose........................................................................................................................ 47
Trombofiliemerkers............................................................................................................. 48
9.2. Genetische trombofilie: loss of function.........................................................................48
Antitrombine...................................................................................................................... 48
2
, Proteïne C........................................................................................................................... 48
Proteïne S.............................................................................................................................. 49
Valkuilen............................................................................................................................. 49
9.3. Genetische trombofilie: Gain of function........................................................................49
F V Leiden mutatie............................................................................................................. 49
Protrombine genmutatie.................................................................................................... 50
Verhoogde F VIII................................................................................................................. 50
Valkuilen............................................................................................................................. 50
9.4. Risico’s........................................................................................................................... 50
9.5. Antifosfolipiden syndroom.............................................................................................. 51
9.6. Richtlijnen trombofilie.................................................................................................... 51
9.7. Antistollingstherapie...................................................................................................... 51
Anti-vitamine K therapie (AVK)........................................................................................... 52
Heparine therapie.............................................................................................................. 52
Directe orale anticoagulantia (DOAC)................................................................................52
Anti-aggregantia................................................................................................................ 53
3
,1. INLEIDING
Hemato-morfologie: vorm en eigenschappen van bloedcellen
Hemocytometrie: chemische en fysische metingen
Hemostase
Immuunhematologie
Moleculaire diagnostiek
2. PRE-ANALYTISCHE FASE
Bloedafname
1) Venapunctie veneus bloed, meestal arm
2) Arteriepunctie arterieel bloed, pols of liesslagader
3) Capillaire punctie capillair bloed, hielprik of vingerprik
Veneus en capillair verschillend door weefselvocht
Anticoagulantia = stolling remmende stof
o Zonder dan bloed stollen
Onderscheid tussen plasma en serum
1) Onstolbaar gemaakt bloed centrifugeren
Pellet = bloecellen
Supernatans = plasma
2) Gestold bloed centrifugeren
Pellet = bloedcellen+ onoplosbaar netwerk van fibrine
Supernatans = serum
BLOED -
BLOEDCELLEN = PLASMA
BLOED - BLOEDCELLEN - FIBRINOGEEN EN STOLLINGSFACTOREN
= SERUM
SERUM = PLASMA - FIBRINOGEEN EN STOLLINGSFACTOREN
Voor centrifugatie Na centrifugatie
4
, Centrifugeren van bloed
= scheiding met centrifugaal kracht en tijd
Centrifugaal kracht = draaisnelheid en afstand van as en rotos
Normaal: 5 min 1500 - 2000 g
Stollingsonderzoek: trombocyten eruit => 15 min 400 g
trombocytenrijk => 5 min 180 g
Chemische anticoagulantia
1. Binding van calciumionen
a. EDTA
b. Citraat
c. Oxalaat
2. Remming van trombine
a. Heparine
EDTA
= ethyleendiamine tetra-azijnzuur
= chelator van Ca2+-ionen
1,5 mg dikaliumzout of trikaliumzout / ml bloed
o K2-EDTA: Lost makkelijk op
Werkt snel
Effect op erytrocyten en trombocyten
o K3-EDTA: Lost minder makkelijk op
Minder snel
Minder effect op volume RBCen TC
Buisje voor bloedafname
o Vooraf EDTA ‘film’ op de wand goed mengen!
o EDTA oplossing is sneller
o Juist volume bloed afnemen juiste concentratie EDTA
Te veel EDTA: ‘creation’ en degeneratie bloedcellen
Gebruiken voor hemocytometrie
Niet gebruiken voor morfologie
Citraat
= trinatriumcitraat
= chelator van Ca2+-ionen
Gebruiken voor sedimentatiesnelheid en stollingstesten
Bestanddeel van ACD-mengsel: onstolbaar maken van bloed voor bloedtransfusie
Altijd in oplossing in de afnamebuis
Finale concentratie BSE: 21 mmol/eindconcentratie
Stollingstesten: 11 mmol/ eindconcentratie
Heparine
Film op wand bloedafnamebuis goed mengen en juiste hoeveelheid bloed !
Niet voor bloeduitstrijkje
Niet voor WBC telling: leukocyten en trombocyten klonteren
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