Subject Index

Affinity chromatography see immunoadsorption Aldehydes see fixation -, see glutaraldehyde Alkaline phosphatase, E. coli 3 Animals 3-4,28 Antibodies, antibovine chymotrypsin 8 -, antiperoxidase 28-29,31,45 -, antirabbit IgG 3-5

Fab and Fc fragments 5,9,20 hyperimmune sera 3-4, 19 IgG and IgM antibodies 4,9,19-20

, isolation 4,8-9,19-20 -, hybrid antibodies 1 Antibody-enzyme conjugates see conjugates Antibody producing cells see lymphoid cells Antibody staining, intracellular antienzyme anti-

bodies 27-28,29,36-42,45-48 - -, penetration of marker enzyme 32,46 Antigen staining, cell surface 12,18

cell suspensions 6-7,11,14 cryostat sections 8, 14, 18, 25 direct method 23 extracellular space 1.2, 18 incubation in conjugates 6, 8, 14, 26 indirect method 23 influence of fixation see fixation intracellular space 12, 23 -25 one-step conjugates 12 penetration of conjugates 14-18

, postembeddingphase 12,18,25 - -, preembedding phase 12,23 - -, tissue fragments 18,24 - -, two-step conjugates 12 - -, ultrathin frozen sections 18, 25 Artifacts, diffusion 3,12,18,24,47 -, non specific adsorption 12,25 -, penetration of conjugates 14,18 -, steric hindrance 18 Autoradiography 28,30-31,42

Bifunctional reagents 20-21 Blast cells see lymphoid cells Bursa of Fabricius, B cells 27

Cell suspensions, enzyme treatment 7 - -, fixation 6, 7 - -, preparation 6, 7 - -, see antigen staining Chromatin, condensation state 48 Chromatography 4-5 Clasmatosis 47 Conjugates, antibody-peroxidase 5,9-11, 19-23 ---, characterization 5-6,9-11,21-23

Ultrastructural Immunoperoxidase Cytochemistry 55

enzyme activity 10,22 immunological activity 10,22

, one-step method 5,10,21-23 - - -, purification 5 - - -, storage 9 ---, two-step method 5,10-11,21-23 Conjugation see conjugates Controls see specificity Copolymers, rabbit IgG-BSA 4,6 Cryostat sections 7,29 Cryo-ultramicrotomy 7 Cytoplasm see lymphoid cells, ultrastructure

Dialdehyde see glutaraldehyde Diaminobenzidine see horseradish peroxidase,

enzymo-cytochemistry Discussion, DNA replication and antibody

synthesis 47-48 -, peroxidase labeled antibodies for immunocyto­

chemical staining 23-26 -, preparation of immunocytochemical

reagents 19-23 - primary immune response to peroxidase

immunization 45,45-47 - secondary immune response to peroxidase

immunization 45,47-48 - ultrastructure of immunocytes 44-48 DNA repair 48 DNA replication 28,42,47-48

f-Amino groups, antibodies and antigens 21,24 - -, peroxidase 21 Electron microscopic techniques 8,29-30 Electrophoresis, enzymo-immuno 6, 28-29 -, SDS 6 Emulsion, Ilford L4 see autoradiography Enzymes, digestion of antibodies 5 - -, cell-coat 7 - immunogens and markers 27-28 -, markers in enzymo-cytochemistry 2-3,20,

27-28 - -, gel filtration 5 Enzymo-cytochemistry see enzymes Ergastoplasm see lymphoid cells, ultrastructure Experimental animals see animals

Fab fragments 5, 9 Fc fragments 5 Ferritin 1,27 Fixation, aldehydes 7,23-24,29 - -, influence on antibody staining 32 ---, antigen staining 14,23-24 -, ethanol-acetic acid 6

56 W. D. KUHLMANN

-, ethanol-ether 29 Fluorescein labeled immune sera Formaldehyde see fixation

Gel diffusion 6 Gel filtration 5 Glucose oxidase, Aspergillus niger 3,5,28 Glutaraldehyde, commercial products 9 -, conjugation with antibodies 5 - -, one-step method 5 - -, two-step method 5 -, fixation 7,24,29 - polymerization 4,7,21 - purity 7 -, vacuum distillation 7 Golgi apparatus see lymphoid cells, ultrastructure

Heavy metals in immunocytochemistry 1 Hemagglutination, passive 29 Historical data, immuno-electron microscopy 1-2 Horseradish peroxidase, «activation» 5

activity 6, 10 conjugation with antibodies 5,9,20-21 enzymo-cytochemistry 3,6,8,29 immunization see immune response immunogen and marker 27-28 purity 3

Hyperimmune sera see antibodies

Illustrations 10-11,13,15,16-17,30-31, 33-35,37-41,43

Immune response, peroxidase 28,31,32-48 ---, primary 28,31,45-47 ---, secondary 28,31,45,47-48 Immune system see lymphoid cells Immunization, Freund's adjuvant 3-4,28,32 -, rabbits 3 - -, booster injection 3 - -, primary 3 -, rats see immune response -, sheep 3-4 Immunoadsorption, cross-linked antigens 4,8, 19-20 -, Sepharose affinity chromatography 4,9,19-20 Immunocompetent cells see lymphoid cells Immunocytes see lymphoid cells Immunocytochemical reagents see conjugates Immunocytochemistry see antibody staining -, see antigen staining Immuno-electron microscopy see antibody staining - - -, see antigen staining - - -, historical data 1-2 Incubation in conjugates see antigen staining Ion exchange chromatography 5 Isolation of antibodies see immunoadsorption

Light microscopy, antiperoxidase cells 31, 45

- -, control of conjugates 6, 11 incubation in conjugates 6, 14,26

, paraffin embedment 6 Lymphocytes see lymphoid cells Lymphoid cells, antibody synthesis 27-28,45-48 - - -, secretion 40, 46, 47

cooperation 27 - -, differentiation 27,32-36,46-48 - -, lymphocytic cell-types 32-42

reticulum cell-types 36-37,44 - -, ultrastructure 32-42,43-48 Lysine see E-amino groups

Macrophages 32 Materials see animals -, see methods Methods, autoradiography 28,30-31

characterization of conjugates 5-6 - conjugation of antibodies with peroxidase 5 - detection of antibodies 6,28-29

electron microscopic techniques 8, 29-31 immunocytochemical procedures 8, 29

- isolation of antibodies 4 - light microscopic cytology 6, 29

preparation of hyperimmune sera 3-4 - -, tissues for immuno-electron microscopy 6-7,29 -, primary immune response 28 -, purification of conjugates 5 -, secondary immune response 28 MOTI cells 36

Nucleolus see lymphoid cells, ultrastructure Nucleoplasm see lymphoid cells, ultrastructure

Papain digestion of antibodies 5,9 Paraffin embedment 6 Paraformaldehyde see fixation Penetration of conjugates see antigen staining -, enzyme markers see antibody staining Perinuclear space see lymphoid cells, ultrastructure Peroxidase, endogenous 12, 25, 32 -, from horseradish see horseradish peroxidase Phytohemagglutinin 47 Plant viruses 1 Plaque-forming test 27 Plasmablasts see lymphoid cells Plasma cells see lymphoid cells Polymerization see copolymers Popliteal lymph nodes for immunocytology 6-7,29 Postfixation 8, 29 Prefixation period 23 Pr!mary immune response, peroxidase see

Immune response Purification of antibodies 4 - -, see immunoadsorption -, conjugates see conjugates

Radial immunodiffusion 6 Radioactivity see autoradiography References 50-54 Replication see DNA replication Results, conjugation of antibodies with

peroxidase 9-11 - DNA replication and antibody synthesis 42 - immuno-electron microscopic cytology 11-18,

31-42 - isolation of antibodies 8-9 - primary immune response 31,32-38 - secondary immune response 31,38-42 - ultrastructure of immunocytes 31-42 Reticulum cells see lymphoid cells Rosette-forming test 27,48 RUSSELL's bodies 36,38,39

Ultrastructural Immunoperoxidase Cytochemistry 57

Secondary immune response, peroxidase see Immune response

Specificity of immunocytochemistry 12, 19, 25-26,49

- -, control reactions 8, 12,25,28,29,32,49 - -, wash procedures 8, 12,26 Summary, Part I 26 --, II 49

Tables 9,14,19,42 Thymidine incorporation see DNA replication Thymus, T cells 27 Tissue fixation see fixation -, sampling 6-7,24-25

Ultrastructure of lymphoid cells see lymphoid cells Ultrathin frozen sections see cryo-ultramicrotomy