Immunohistochemistry and Immunocytochemistry: Methods and Protocols

284 Pages·2021·8.721 MB·English

by  Luis Del Valle

Methods in
Molecular Biology 2422
Luis Del Valle Editor
Immuno histo-
chemistry
and Immunocyto-
chemistry
Methods and Protocols
M M B
ETHODS IN OLECULAR IO LO GY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire, UK
Forfurther volumes:
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For over 35 years, biological scientists have come to rely on the research protocols and
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Immunohistochemistry
and Immunocytochemistry
Methods and Protocols
Edited by
Luis Del Valle
Department of Pathology and Medicine & Louisiana Cancer Research Center, Louisiana State
University Health Sciences Center, New Orleans, LA, USA
Editor
LuisDelValle
DepartmentofPathologyandMedicine
&LouisianaCancerResearchCenter
LouisianaStateUniversityHealthSciencesCenter
NewOrleans,LA,USA
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-0716-1947-6 ISBN978-1-0716-1948-3 (eBook)
https://doi.org/10.1007/978-1-0716-1948-3
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Preface
Theuseoflabeled antibodiesfor thespecificdetection andaccurate localizationofcellular
proteins and structures was fathered by A.H. Coons in the 1940s, but it was not until the
early 1970s whenit exploded into a widespread technique. Since thenimmunohistochem-
istry and immunocytochemistry have become indispensable tools in diagnostic pathology
departments and essential investigative techniques in research laboratories. Labeling with
antibodies issuch a powerful and versatile tool; it can beapplied to cellcultures, frozenor
fixed tissue, from single cell microorganisms to plants, pathogens, experimental animal
models all the way to human tissues, and to a variety of techniques, from classic immuno-
histochemistry to novel assays, such as in situ proximity ligation (Duo Link). However,
becauseoftheimportanceoftheanswersitprovides,theneedforaccurateandreproducible
results is of crucial importance, and small variations in protocols can result in unreliable
results.Withthisinmind,wehavecompiledagroupofexpertsinthefieldtoprovidetheir
knowledge in the different aspects of the process and in the different techniques in which
immunoassays are utilized. Each chapter is written by an expert in the field and includes a
protocolusedintheirownlaboratoriesaswellasdetailedinformationoneachsubject.
This book offers a comprehensive reference guide to anyone interested in the applica-
tions of labeled antibodies, from novice graduate students who wish to understand the
theory and practice of immunohistochemistry, immunocytochemistry, and fluorescent
methodologies to the experienced immunochemist, pathologist, or researcher who may
wishtofurtheroptimizetheirexistingprotocols.
NewOrleans,LA,USA LuisDelValle
v
Contents
Preface ..................................................................... v
Contributors................................................................. ix
1 IntroductiontoImmunohistochemistry:FromtoEvolvingScience
toTimelessArt ......................................................... 1
LuisDelValle
2 ImmunohistochemistryinHistoricalPerspective:KnowingthePast
toUnderstandthePresent ............................................... 17
CarlosOrtizHidalgo
3 SamplePreparationandFixationforHistologyandPathology................ 33
EnriqueAgustı´nSampedro-Carrillo
4 TissueProcessing ....................................................... 47
SarahJ.AzizandCadyE.Zeman-Pocrnich
5 AntigenRetrievalandSignalAmplification................................. 65
LoganDunkenbergerandLuisDelValle
6 PrimaryAntibodySelectionandBlockingTechniques
forImmunohistochemistry............................................... 75
LoriA.PetonandMiguelReyes-Mu´gica
7 ProbesforFluorescentVisualizationofSpecificCellularOrganelles........... 85
TimothyPaulFoster
8 EnzymaticImmunohistochemistry........................................ 125
RodolfoHenrichLobo
9 FluorescentImmunohistochemistry....................................... 131
VanessaMoreno,ElizabethA.Smith,andSergioPin˜a-Oviedo
10 DoubleLabelingFluorescentImmunocytochemistry........................ 147
MonikaRakandKrzysztofReiss
11 MultiplexingandSpectralMicroscopy..................................... 163
LoganDunkenberger,AdrianaZapata,andLuisDelValle
12 FluorescentInSituHybridization(FISH) ................................. 179
JeanetteM.Ramos
13 ProximityLigationAssay(PLA) .......................................... 191
MuhammadS.Alam
14 Immunohistochemistry,QualityControl,andPrinciplesofValidation
intheCentralNervousSystem ........................................... 203
Ver(cid:1)onicaVelasco-Vales,DannySoria-Ce´spedes,
TeresaCristinaCuesta-Mejı´as,andNormaCeciliaPadr(cid:1)on-Pe´rez
15 CultureandPhenotypingofGlialCellCultures,
Gliospheres,andNeurospheres........................................... 217
LuisDelValle,KrzysztofReiss,andAmandaParkerStruckhoff
vii
viii Contents
16 ER+BreastCancerMammosphereFormationandAnalysis .................. 233
CieraS.Singleton,LeoLi-YingChan,KelseyJ.McCulley,
SarahL.Kessel,LuisDelValle,andJudyS.Crabtree
17 DetectionofCirculatingandTissueMyeloid-DerivedSuppressor
Cells(MDSC)byFlowCytometry........................................ 247
MariaDulfarySanchez-Pino
18 CometAssayfor theDetectionofSingleandDouble-Strand
DNABreaks............................................................ 263
LoganDunkenberger,KrzysztofReiss,andLuisDelValle
19 Fluorescent-ActivatedCellSorting(FlowCytometry)....................... 271
LuisF.Carrillo
Index ...................................................................... 283
Contributors
MUHAMMADS.ALAM • LaboratoryofImmuneCellBiology,NationalCancerInstitute,
NationalInstitutesofHealth,Bethesda,MD,USA
SARAHJ.AZIZ • DepartmentofPathologyandLaboratoryMedicine,WesternUniversityand
LondonHealthSciencesCentre,London,ON,Canada
LUISF.CARRILLO • DepartmentofPathology,CollegeofMedicine,UniversityofArkansasfor
MedicalSciences,LittleRock,AR,USA
LEOLI-YINGCHAN • DepartmentofAdvancedTechnologyR&D,NexcelomBioscienceLLC,
Lawrence,MA,USA
JUDY S.CRABTREE • DepartmentofGenetics,LouisianaStateUniversityHealthSciences
Center,NewOrleans,LA,USA
TERESACRISTINACUESTA-MEJI´AS • DepartamentodePatologı´a,CentroMedicoABC,
CiudaddeMe´xico,Mexico;DepartamentodePatologı´a,HospitalJua´rezdeMe´xico,
CiudaddeMe´xico,Mexico
LUIS DELVALLE • DepartmentofPathologyandMedicine&LouisianaCancerResearch
Center,LouisianaStateUniversityHealthSciencesCenter,NewOrleans,LA,USA
LOGANDUNKENBERGER • NeurologicalCancerResearch,LouisianaCancerResearchCenter,
LouisianaStateUniversityHealthSciencesCenter,NewOrleans,LA,USA
TIMOTHYPAULFOSTER • DepartmentofMicrobiology,Immunology&Parasitology,
LouisianaStateUniversityHealthSciencesCenter,NewOrleans,LA,USA
RODOLFOHENRICHLOBO • DepartmentofPathology,CollegeofMedicine,Universityof
ArkansasforMedicalSciences,LittleRock,AR,USA
SARAH L.KESSEL • DepartmentofAdvancedTechnologyR&D,NexcelomBioscienceLLC,
Lawrence,MA,USA
KELSEYJ.MCCULLEY • DepartmentofAdvancedTechnologyR&D,NexcelomBioscience
LLC,Lawrence,MA,USA
VANESSAMORENO • ArkanaLaboratories,LittleRock,AR,USA
CARLOSORTIZHIDALGO • DepartmentofAnatomicPathology,Fundaci(cid:1)onClı´nicaMe´dica
Sur,CiudaddeMe´xico,Mexico;DepartamentodeBiologı´aCelular yTisular,Facultadde
Medicina,UniversidadPanamericana,CiudaddeMe´xico,Mexico
NORMACECILIAPADRO´N-PE´REZ • DepartamentodePatologı´a,CentroMedicoABC,Ciudad
deMe´xico,Mexico
AMANDAPARKERSTRUCKHOFF • NeurologicalCancerResearch,LouisianaCancerResearch
Center,LouisianaStateUniversityHealthSciencesCenter,NewOrleans,LA,USA
LORI A.PETON • DepartmentofPathologyandLaboratoryMedicine,UPMCChildren’s
HospitalofPittsburgh,Pittsburgh,PA,USA;UPMCSchoolofHistotechnology,Pittsburgh,
PA,USA
SERGIOPIN˜A-OVIEDO • DepartmentofPathology,UniversityofArkansasforMedical
Sciences,LittleRock,AR,USA
MONIKARAK • DepartmentofCellBiology,FacultyofBiochemistry,Biophysicsand
Biotechnology,JagiellonianUniversityinKrak(cid:1)ow,Krak(cid:1)ow,Poland;Departmentsof
Medicine&NeurologicalCancerResearch,LouisianaCancerResearchCenter,Louisiana
StateUniversityHealthSciencesCenter,NewOrleans,LA,USA
ix
x Contributors
JEANETTEM.RAMOS • DepartmentofPathologyandLaboratoryServices,Universityof
ArkansasforMedicalSciences,LittleRock,AR,USA
KRZYSZTOF REISS • DepartmentsofMedicine&NeurologicalCancerResearch,Louisiana
CancerResearchCenter,LouisianaStateUniversityHealthSciencesCenter,NewOrleans,
LA,USA
MIGUELREYES-MU´GICA • DepartmentofPathologyandLaboratoryMedicine,UPMC
Children’sHospitalofPittsburgh,Pittsburgh,PA,USA;UniversityofPittsburghSchoolof
Medicine,Pittsburgh,PA,USA
ENRIQUEAGUSTI´NSAMPEDRO-CARRILLO • DepartamentodeBiologı´aCelular yTisular,
FacultaddeMedicina,UniversidadNacionalAut(cid:1)onomadeMe´xico,Me´xicoD.F.,Mexico
MARIADULFARYSANCHEZ-PINO • DepartmentofInterdisciplinaryOncologyand
DepartmentofGenetics,LouisianaStateUniversityHealthSciencesCenter,NewOrleans,
LA,USA
CIERAS.SINGLETON • DepartmentofGenetics,LouisianaStateUniversityHealthSciences
Center,NewOrleans,LA,USA
ELIZABETHA.SMITH • ArkanaLaboratories,LittleRock,AR,USA
DANNYSORIA-CE´SPEDES • DepartamentodePatologı´a,CentroMedicoABC,Ciudadde
Me´xico,Mexico
VERO´NICAVELASCO-VALES • DepartamentodePatologı´a,HospitalA´ngelesLomas,Estadode
Me´xico,Mexico
ADRIANA ZAPATA • LouisianaCancerResearchCenter,LouisianaStateUniversityHealth,
NewOrleans,LA,USA
CADYE.ZEMAN-POCRNICH • DepartmentofPathologyandLaboratoryMedicine,Western
UniversityandLondonHealthSciencesCentre,London,ON,Canada