Methoden in biomedisch
onderzoek 2
Table of Contents
VOORWOORD AUTEUR..................................................................................4
AFKORTINGENLIJST......................................................................................4
1. INLEIDING VAK................................................................................... 6
2. VRAAGSTELLING EN ONDERZOEKSMETHODIEK..................................... 6
3. ANALYSE VAN RNA.............................................................................. 6
WERKEN MET RNA, BEWARING.......................................................................7
RNA EXTRAHEREN......................................................................................7
RNA QUANTIFICEREN...................................................................................8
RNA CONCENTREREN....................................................................................................... 9
RNA IN VITRO AANMAKEN...........................................................................10
REVERSE TRANSCIPTION..............................................................................10
RT-REACTIE: AANMAAK CDNA (COPY OF COMPLEMENT)........................................................10
CDNA KLONEREN.......................................................................................................... 11
RACE (ZWARTE DEEL GEKENDE SEQ, GRIJS ONBEKEND)........................................................12
RNA ANALYSEREN.....................................................................................12
NORTHERN BLOT........................................................................................................... 13
KWANTITATIEVE TOEPASSINGEN: RT-QPCR EN DDPCR..........................................................13
IN-SITU METHODEN........................................................................................................ 15
4. ANALYSE VAN EIWITTEN.................................................................... 17
1) ISOLEREN VAN EIWITTEN.........................................................................17
2) AANMAKEN VAN EIWITTEN!......................................................................17
1. CHEMISCHE SYNTHESE IN VITRO................................................................................... 17
2. ENZYMATISCHE SYNTHESE IN VITRO...............................................................................17
3. IN VIVO EXPRESSIE SYSTEMEN...................................................................................... 17
4. BEREIDING ANTISTOFFEN............................................................................................. 17
3) CONCENTRATIEBEPALING.........................................................................18
1. UV-ABSORPTIE.......................................................................................................... 18
2. COLORIMETRISCHE METHODEN..................................................................................... 18
4) CONCENTREREN VAN EIWITTEN..................................................................18
,1. ZOUTPRECIPITATIE (UITZOUTEN)................................................................................... 18
2. CENTRIFUGALE FILTERS............................................................................................... 19
5) KWANTIFICEREN/IDENTIFICEREN VAN SPECIFIEKE EIWITTEN................................19
1. WESTERN BLOTTING................................................................................................... 19
2. ANTILICHAAM-MICROARRAYS........................................................................................ 19
3. FLOW CYTOMETRIE..................................................................................................... 19
4. ELISA.................................................................................................................... 19
5. RADIOIMMUNOASSAY................................................................................................. 19
6. IMMUNOHISTOCHEMIE................................................................................................ 20
5. MICROSCOPIE................................................................................... 20
1) FLUORESCENTIEMICROSCOPIE....................................................................20
2) BEELDANALYSE.....................................................................................22
3) SUPERRESOLUTIE MICROSCOPIE.................................................................23
4) ELEKTRONENMICROSCOPIE.......................................................................26
5) SCANNING PROBE MICROSCOPIE...............................................................28
6. ELEKTROFYSIOLOGISCHE METINGEN.................................................. 28
1) BASISBEGINSELEN.................................................................................28
2) EQUIVALENTE CIRCUITS...........................................................................29
3) INTRACELLULAIRE MEETTECHNIEKEN............................................................29
4) EXTRACELLULAIRE MEETTECHNIEKEN...........................................................30
5) OPTISCHE MEETTECHNIEKEN.....................................................................31
7. IN-VIVO METINGEN BIJ MENS.............................................................. 32
0. INLEIDING...........................................................................................32
1. ULTRASONE BEELDVORMING (US)..............................................................32
1) KARAKTERISTIEKEN VAN GELUID EN GELUIDSGOLVEN.........................................................32
2) INTERACTIES TUSSEN ULTRASONE GOLVEN EN MATERIE......................................................32
3) APPARATUUR............................................................................................................ 33
4) ACQUISITIE (MEETPROCES).......................................................................................... 33
5) DOPPLER-ECHOGRAFIE............................................................................................... 33
6) AFSTANDSMETINGEN.................................................................................................. 33
7) CONTRASTMIDDEN..................................................................................................... 33
8) BIOLOGISCHE EFFECTEN............................................................................................. 33
2. MRI..................................................................................................34
1) INLEIDING................................................................................................................ 34
2) RESONANTIE............................................................................................................ 34
3) T1 EN T2 RELAXATIEPARAMETERS................................................................................35
,4) BASIS ACQUISITIEPARAMETERS..................................................................................... 35
5) SPIN ECHO (SE)....................................................................................................... 35
6) INVERSION RECOVERY SPIN ECHO SEQUENTIE (IR SE).......................................................35
7) GRADIENT ECHO (GE)............................................................................................... 36
8) SIGNAALLOCALISATIE.................................................................................................. 36
9) K-RUIMTE................................................................................................................. 36
10) SPATIALE RESOLUTIE EN CONTRAST............................................................................. 36
11) ANGIOGRAFIE (MRA)............................................................................................... 37
12. FUNCTIONELE FMRI................................................................................................. 37
13. DIFFUSIE MRI......................................................................................................... 37
3. INTERACTIE STRALING-MATERIE..................................................................37
1) EMS...................................................................................................................... 37
2) RAYLEIGH-VERSTROOIING............................................................................................ 37
3) COMPTON-VERSTROOIING........................................................................................... 38
4) FOTO-ELEKTRISCH EFFECT........................................................................................... 38
5) PAARPRODUCTIE....................................................................................................... 38
6) DOMINANTE INTERACTIE............................................................................................. 38
7) LINEAIRE ATTENUATIECOËFFICIËNT Μ.............................................................................38
4. X-STRALEN BEELDVORMING......................................................................39
1) INTRODUCTIE............................................................................................................ 39
2) X-STRALEN PRODUCTIE............................................................................................... 39
3) X-STRALEN DETECTIE................................................................................................. 39
4) FYSISCH PRINCIPE CT EN BEELDRECONSTRUCTIE.............................................................39
5) CT ACQUISITIES........................................................................................................ 40
6) CT ANGIOGRAFIE....................................................................................................... 40
7) CT PERFUSIE............................................................................................................ 40
8) CARDIO CT.............................................................................................................. 40
9) DUAL SOURCE CT..................................................................................................... 40
10) PHOTON-COUNTING CT............................................................................................ 40
11) VOOR-/NADELEN CT................................................................................................ 41
5. PRODUCTIE RADIO-ISOTOPEN....................................................................41
1) INLEIDING................................................................................................................ 41
2) RADIOACTIEF VERVAL................................................................................................. 41
3) PRODUCTIE RADIONUCLIDEN........................................................................................ 42
6. SPECT..............................................................................................42
INLEIDING.................................................................................................................... 42
GAMMA-CAMERA........................................................................................................... 42
COMPUTER TOMOGRAFIE................................................................................................. 43
PARTIEEL VOLUME EFFECT............................................................................................... 43
ATTENUATIECORRECTIE................................................................................................... 43
VERSTROOIINGSCORRECTIE.............................................................................................. 43
TOEPASSINGEN............................................................................................................. 43
NIEUWE ONTWIKKELINGEN............................................................................................... 43
VOOR-/NADELEN SPECT................................................................................................ 43
, 7. PET..................................................................................................43
INLEIDING.................................................................................................................... 44
COÏNCIDENTIE-DETECTIE................................................................................................. 44
DETECTOREN................................................................................................................ 44
CORRECTIES................................................................................................................. 45
COMPUTER TOMOGRAFIE; RECONSTRUCTIE..........................................................................45
KINETISCHE MODELLERING.............................................................................................. 46
NIEUWE ONTWIKKELINGEN............................................................................................... 46
VOOR-/NADELEN........................................................................................................... 46
8. RADIOTHERAPIE PRINCIPES.......................................................................47
INTERACTIE DEELTJES-MATERIE.......................................................................................... 47
STRALINGSBIOLOGIE....................................................................................................... 47
RADIOTHERAPIE............................................................................................................ 47
4. PROTONTHERAPIE...................................................................................................... 47
9. RADIOPROTECTIE...................................................................................47
INLEIDING.................................................................................................................... 47
DETERMINISTISCHE EN STOCHASTISCHE EFFECTEN................................................................48
EFFECTIEVE DOSIS......................................................................................................... 48
BESCHERMING.............................................................................................................. 48
RISICO CLASSIFICATIE..................................................................................................... 48
(RICHTLIJNEN)..........................................................................................48
8. RADIO-ISOTOOPTECHNIEKEN LABO.................................................... 49
KLEINE PROEFDIEREN..................................................................................................... 49
RADIOIMMUNOASSAY..................................................................................................... 49
Voorwoord auteur
- Zowel notities bij slides als full blown samengevatte stukken
- Screenshots van slides heb ik verwijderd (copyright?) maar in de plaats
staat nog een vermelding [tussen vierkante haken]
- (onvolledige) wetenschappelijke afkortingenlijst, verder nog
onconventionele afkortingen zoals eiwit ewt, hoeveelheid hvlhd,
afhankelijk van afhv, …
Stuur gerust bij onduidelijkheden.
Afkortingenlijst
βME: Bèta-MercaptoEthanol
BSE: BackScattered e-
CT: Computed Tomography scan
onderzoek 2
Table of Contents
VOORWOORD AUTEUR..................................................................................4
AFKORTINGENLIJST......................................................................................4
1. INLEIDING VAK................................................................................... 6
2. VRAAGSTELLING EN ONDERZOEKSMETHODIEK..................................... 6
3. ANALYSE VAN RNA.............................................................................. 6
WERKEN MET RNA, BEWARING.......................................................................7
RNA EXTRAHEREN......................................................................................7
RNA QUANTIFICEREN...................................................................................8
RNA CONCENTREREN....................................................................................................... 9
RNA IN VITRO AANMAKEN...........................................................................10
REVERSE TRANSCIPTION..............................................................................10
RT-REACTIE: AANMAAK CDNA (COPY OF COMPLEMENT)........................................................10
CDNA KLONEREN.......................................................................................................... 11
RACE (ZWARTE DEEL GEKENDE SEQ, GRIJS ONBEKEND)........................................................12
RNA ANALYSEREN.....................................................................................12
NORTHERN BLOT........................................................................................................... 13
KWANTITATIEVE TOEPASSINGEN: RT-QPCR EN DDPCR..........................................................13
IN-SITU METHODEN........................................................................................................ 15
4. ANALYSE VAN EIWITTEN.................................................................... 17
1) ISOLEREN VAN EIWITTEN.........................................................................17
2) AANMAKEN VAN EIWITTEN!......................................................................17
1. CHEMISCHE SYNTHESE IN VITRO................................................................................... 17
2. ENZYMATISCHE SYNTHESE IN VITRO...............................................................................17
3. IN VIVO EXPRESSIE SYSTEMEN...................................................................................... 17
4. BEREIDING ANTISTOFFEN............................................................................................. 17
3) CONCENTRATIEBEPALING.........................................................................18
1. UV-ABSORPTIE.......................................................................................................... 18
2. COLORIMETRISCHE METHODEN..................................................................................... 18
4) CONCENTREREN VAN EIWITTEN..................................................................18
,1. ZOUTPRECIPITATIE (UITZOUTEN)................................................................................... 18
2. CENTRIFUGALE FILTERS............................................................................................... 19
5) KWANTIFICEREN/IDENTIFICEREN VAN SPECIFIEKE EIWITTEN................................19
1. WESTERN BLOTTING................................................................................................... 19
2. ANTILICHAAM-MICROARRAYS........................................................................................ 19
3. FLOW CYTOMETRIE..................................................................................................... 19
4. ELISA.................................................................................................................... 19
5. RADIOIMMUNOASSAY................................................................................................. 19
6. IMMUNOHISTOCHEMIE................................................................................................ 20
5. MICROSCOPIE................................................................................... 20
1) FLUORESCENTIEMICROSCOPIE....................................................................20
2) BEELDANALYSE.....................................................................................22
3) SUPERRESOLUTIE MICROSCOPIE.................................................................23
4) ELEKTRONENMICROSCOPIE.......................................................................26
5) SCANNING PROBE MICROSCOPIE...............................................................28
6. ELEKTROFYSIOLOGISCHE METINGEN.................................................. 28
1) BASISBEGINSELEN.................................................................................28
2) EQUIVALENTE CIRCUITS...........................................................................29
3) INTRACELLULAIRE MEETTECHNIEKEN............................................................29
4) EXTRACELLULAIRE MEETTECHNIEKEN...........................................................30
5) OPTISCHE MEETTECHNIEKEN.....................................................................31
7. IN-VIVO METINGEN BIJ MENS.............................................................. 32
0. INLEIDING...........................................................................................32
1. ULTRASONE BEELDVORMING (US)..............................................................32
1) KARAKTERISTIEKEN VAN GELUID EN GELUIDSGOLVEN.........................................................32
2) INTERACTIES TUSSEN ULTRASONE GOLVEN EN MATERIE......................................................32
3) APPARATUUR............................................................................................................ 33
4) ACQUISITIE (MEETPROCES).......................................................................................... 33
5) DOPPLER-ECHOGRAFIE............................................................................................... 33
6) AFSTANDSMETINGEN.................................................................................................. 33
7) CONTRASTMIDDEN..................................................................................................... 33
8) BIOLOGISCHE EFFECTEN............................................................................................. 33
2. MRI..................................................................................................34
1) INLEIDING................................................................................................................ 34
2) RESONANTIE............................................................................................................ 34
3) T1 EN T2 RELAXATIEPARAMETERS................................................................................35
,4) BASIS ACQUISITIEPARAMETERS..................................................................................... 35
5) SPIN ECHO (SE)....................................................................................................... 35
6) INVERSION RECOVERY SPIN ECHO SEQUENTIE (IR SE).......................................................35
7) GRADIENT ECHO (GE)............................................................................................... 36
8) SIGNAALLOCALISATIE.................................................................................................. 36
9) K-RUIMTE................................................................................................................. 36
10) SPATIALE RESOLUTIE EN CONTRAST............................................................................. 36
11) ANGIOGRAFIE (MRA)............................................................................................... 37
12. FUNCTIONELE FMRI................................................................................................. 37
13. DIFFUSIE MRI......................................................................................................... 37
3. INTERACTIE STRALING-MATERIE..................................................................37
1) EMS...................................................................................................................... 37
2) RAYLEIGH-VERSTROOIING............................................................................................ 37
3) COMPTON-VERSTROOIING........................................................................................... 38
4) FOTO-ELEKTRISCH EFFECT........................................................................................... 38
5) PAARPRODUCTIE....................................................................................................... 38
6) DOMINANTE INTERACTIE............................................................................................. 38
7) LINEAIRE ATTENUATIECOËFFICIËNT Μ.............................................................................38
4. X-STRALEN BEELDVORMING......................................................................39
1) INTRODUCTIE............................................................................................................ 39
2) X-STRALEN PRODUCTIE............................................................................................... 39
3) X-STRALEN DETECTIE................................................................................................. 39
4) FYSISCH PRINCIPE CT EN BEELDRECONSTRUCTIE.............................................................39
5) CT ACQUISITIES........................................................................................................ 40
6) CT ANGIOGRAFIE....................................................................................................... 40
7) CT PERFUSIE............................................................................................................ 40
8) CARDIO CT.............................................................................................................. 40
9) DUAL SOURCE CT..................................................................................................... 40
10) PHOTON-COUNTING CT............................................................................................ 40
11) VOOR-/NADELEN CT................................................................................................ 41
5. PRODUCTIE RADIO-ISOTOPEN....................................................................41
1) INLEIDING................................................................................................................ 41
2) RADIOACTIEF VERVAL................................................................................................. 41
3) PRODUCTIE RADIONUCLIDEN........................................................................................ 42
6. SPECT..............................................................................................42
INLEIDING.................................................................................................................... 42
GAMMA-CAMERA........................................................................................................... 42
COMPUTER TOMOGRAFIE................................................................................................. 43
PARTIEEL VOLUME EFFECT............................................................................................... 43
ATTENUATIECORRECTIE................................................................................................... 43
VERSTROOIINGSCORRECTIE.............................................................................................. 43
TOEPASSINGEN............................................................................................................. 43
NIEUWE ONTWIKKELINGEN............................................................................................... 43
VOOR-/NADELEN SPECT................................................................................................ 43
, 7. PET..................................................................................................43
INLEIDING.................................................................................................................... 44
COÏNCIDENTIE-DETECTIE................................................................................................. 44
DETECTOREN................................................................................................................ 44
CORRECTIES................................................................................................................. 45
COMPUTER TOMOGRAFIE; RECONSTRUCTIE..........................................................................45
KINETISCHE MODELLERING.............................................................................................. 46
NIEUWE ONTWIKKELINGEN............................................................................................... 46
VOOR-/NADELEN........................................................................................................... 46
8. RADIOTHERAPIE PRINCIPES.......................................................................47
INTERACTIE DEELTJES-MATERIE.......................................................................................... 47
STRALINGSBIOLOGIE....................................................................................................... 47
RADIOTHERAPIE............................................................................................................ 47
4. PROTONTHERAPIE...................................................................................................... 47
9. RADIOPROTECTIE...................................................................................47
INLEIDING.................................................................................................................... 47
DETERMINISTISCHE EN STOCHASTISCHE EFFECTEN................................................................48
EFFECTIEVE DOSIS......................................................................................................... 48
BESCHERMING.............................................................................................................. 48
RISICO CLASSIFICATIE..................................................................................................... 48
(RICHTLIJNEN)..........................................................................................48
8. RADIO-ISOTOOPTECHNIEKEN LABO.................................................... 49
KLEINE PROEFDIEREN..................................................................................................... 49
RADIOIMMUNOASSAY..................................................................................................... 49
Voorwoord auteur
- Zowel notities bij slides als full blown samengevatte stukken
- Screenshots van slides heb ik verwijderd (copyright?) maar in de plaats
staat nog een vermelding [tussen vierkante haken]
- (onvolledige) wetenschappelijke afkortingenlijst, verder nog
onconventionele afkortingen zoals eiwit ewt, hoeveelheid hvlhd,
afhankelijk van afhv, …
Stuur gerust bij onduidelijkheden.
Afkortingenlijst
βME: Bèta-MercaptoEthanol
BSE: BackScattered e-
CT: Computed Tomography scan
