9 Subject index

Acridine fluorescence 66Aldehyde-forrnazan-technique 54Amidrazones, from forrnazans 13, 14Amines, from forrnazans 13Antimycin A 58p-Arylazoarylhydrazones 11Arylazoforrnazans 12, 13Atypical forrnazans- biological significance 64, 65- birefringence 65, 66- and cell state 65- dichroism 65- enzymatic formation 65- and lipids 65- optical characteristics 66- and type of injury 65

Bathochromic shift 26, 36, 37, 40, 422-Benzothiazolyl-3-phthalhydrazidyl-5-sty-

ryl-tetrazolium chloride, BPST see BPSTBis(arylazo)methan derivatives 12Blue tetrazolium, see BTBPST salt- lipophilia 45- reducibility 23, 53, 55- reduction and oxygen 56- spectrum 24- substantivity 62- use in electron microscopy 42, 45BPST formazan- localizing potentials 35, 62- metal chelation 9- osmiophilic property 45, 46- osmium black of 46- spectrum 25, 27BT salt 37, 58- in electron microscopy 44- half-wave potential 22- reducibility 23, 53, 54, 55, 57- TLC 17- UV spectrum 24BT formazan- TLC 17- visible spectrum 25

Cell injuries- detection by nitrotetrazolium salts 63, 64"Conjugated bond number" 48, 49Correlations between structure of- forrnazans and color 40- forrnazans and localizing qualities 41- formazans and substantivity 38- tetrazolium salts and histochemical use 1, 17,

18, 35, 66- tetrazolium salts and reducibility 38- tetrazolium salts and substantivity 383-Cyan-l,5-diarylformazans 475-Cyan-2,3-di(p-tolyl)-2H-tetrazolium

chloride, CTCas fluorescent tetrazolium salt 47

Cyano group and fluorescencein formazans 47

Cytochrome c 58, 60

Detergents and tetrazolium reduction 23, 53,57,61

Diaphorase and tetrazolium reducibility 23Diformazans- contamination with monoformazans 11, 15- localization qualities 33- optical characteristics 24 et seq. 25, 27, 29, 33- physical properties 6, 9, 10- preparation 11- purification from by products 11- TLC 17- valuation 623-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl­

2H-tetrazolium bromide, MIT 10, 54, 58,62

- half-wave potential 22- and oxidative phosphorylation 50- reducibility 22, 53- spectrum 24- role of substantivity 20- TLC 17- MIT formazan- color in solvents 26- in electron microscopy 44- metal chelation 9, 44- spectrum 25

- spectrum of Co-chelat 25- TLC 17p-Dinitrotetrazolium chloride, p-DNTIC- "nitrosensitivity" 37, 64- "nitrosensitivity" and color 64- quantification. 66- reducibility 23, 53- UV spectrum 24p-DNTIC formazan- "aci-form" spectrum 33- visible spectrum 25, 27a-Dinitrotetrazolium violet, a-DNTV- "nitrosensitivity" 64- reducibility 23a-DNTV formazan- "aci-form" spectrum 33- visible spectrum 25- UV spectrum 241,5-Di(phenyl)-3-H-formazan 271,5-Di(phenyl)-3-methyl-formazan 27Dismutation- of superoxide(o Oz) radicals 60- of tetrazolinyls (0 TS) 52, 62Distyryl-nitroblue tetrazolium 44Ditetrazolium salts- contamination 15- half-wave potentials 22- heterogeneity of reaction product 54, 62- histochemical advantages 62- mechanism of reduction 51, 52- optical characteristics 24- purification from mono-salts 14- TLC 16, 17- TLC of half-reduction products 17- N-N'-type 5, 6, 37- C-C' -type 5, 6, 37, 42- valuation of the "red" and "blue" products

53,54,62

Enzyme inhibition- by metal ions 35- by tetrazolium salts 35

Fluorescent formazans and tetrazolium salts46,47

Formazans- "aci-form" 29, 31, 32- amino-imino-tautomerism 9, 26, 36

Subject index' 81

- amphoteric character 31- analytical chemistry 15, 16, 17- as anions 30, 31- biaryl-substituted 11, 15- biological uses 9- "blue" form 7, 54, 62, 64- as by-products in diformazans 11, 15- chemical and physical properties 8, 9, 10, 12,

15- decomposition 11- diffusion in tissues 20, 26- as dipole 32- elution from sections 20- extinction coefficients 25, 27, 33- formation from tetrazolinyls 28, 51, 52- formation from tetrazolium salts 4,7,18,21,

51, 52, 54, 59, 60- halfreduction products of ditetrazolium

salts, see half-formazans- halochromism 29, 30- Hansch-n-values 48, 49- effects of hetaryl groups 8, 9, 10, 13- heterogeneity of colors 35- history 1, 6- hydrogen bonds 7, 8, 9, 10,20,28- IR-spectroscopy 17- isomeric forms 6, 7, 8, 9, 30, 55- light effects on structure 30, 35, 55- lipophilia 34, 48- localizing qualities 18, 33, 34, 35, 62- mesomerism and tautomerism 7, 8, 9- metal chelation 8, 9, 10, 38, 55- microdensitometry 26, 28, 66- numbering system 6- negative metachromasy 32, 37, 65- nitrodichroism 29,31, 32, 33, 37, 42- nitrosensitivity 37, 63, 64, 65- optical anisotropy 37- optical characteristics 15, 24 et seq., 33, 40- properties and parameters for histochemical

use 17, 18- photochromism 28, 29- photooxidation 28, 29- properties in tissues 18,32,33,34, 55- purity and purification 11, 15, 16, 17,36- quantifying 15, 26, 28- as quasiaromatic rings 7, 8- "red" form 7, 8, 9,25,27,28,30,31,55,62,

64

82 . Subject index

- reduction beyond the formazan stage 13, 14,21,22

- as resonance hybrid 8- solubilities 10, 18, 20, 48, 49, 55- solvatochromism 29, 31, 42- structure 6, 7, 8, 9, 10,26,31- substantivity 18- synthesis 10, 11, 12, 13- thermochromism 29, 31, 42- time stability 35- TLC 16, 17- transarylation 12- types 6,7- "yellow" form 7, 27, 29, 30, 31, 55

Glucose-6-phosphate dehydrogenase 53

Half-formazanes- formation 51, 52, 54, 60- localizing qualities 34, 35- optical data 25, 26, 27- properties 51, 52, 54, 55, 62- significance 54, 55, 62- solubilities 34, 35- stability 51, 52- structure 51, 52Hammett-(J-constants 48, 49Hansch-n-values 48, 49, 53Histochemical qualities and histochemical de-

sign 35, 36, 37, 38, 39, 40, 41, 42Histochemical result- interpretation 15- quantifying 15Hydrazidines 13, 14

Inhibitors of respiratory chain 582-Iodophenyl-3-p-nitrophenyl-5-phenyl-2-H-

tetrazolium chloride INT 58, 62- in electron microscopy 44- half-wave potential 22- and oxidative phosphorylation 50- reducibility 23- TLC 17- UV spectrum 24INT formazan- spectrum of "aci-form" 33- TLC 17- visible spectrum 25

Isobestic wavelength 26Iso-nitrotetrazolium salts 37, 64

Mediators 53, 56, 58, 59, 60, 61M-effects 38, 39, 40, 41, 48, 51Menadion 58o-Methyl-lTC VII, 22Microdensitometric techniques 26Monoamino oxidase, MAO 50MPMS 56MSAO-technique 66Monoformazans- diaryl-substituted 11, 15, 62- discrimination in TLC 15, 16- from monotetrazolium salts 62- optical data 24, 25, 27, 33, 65, 66Monotetrazolium salts- advantages in histochemical use 62- histochemical applications 62- contaminating ditetrazolium salts 15, 62- half-wave potentials 22- isomeric forms in TLC 16, 17- optical data 24- preparation 62- significance of reduction products 55- TLC 16,17

NADH-PMS test 23NBT 18, 42, 52, 58, 61, 62, 63- in electron microscopy 44- formation of mono-formazan 11- half-wave potential 22- iodine derivatives 44- isomeric forms 16- localizing qualities 35, 63- in oxidative phosphorylation 50- acetone preincubation 20- redox potential 21- reducibility 23, 53, 54, 55- reduction and oxygen 56- reduction by oxygen radical 62- as standard indicator 18- synthesis 11- TLC 16, 17- UV spectra 24NBT formazan- "blue granula" 63- "nitrosensitivity" 64- spectrum 25, 27

- spectrum of "aci-form" 33- TLC 16, 17NBTmono, 2-p-nitrophenyl-3-(3,3' -dimethoxy­

4-biphenylylene)-5-phenyl-2H-tetrazoliumchloride 22

NBT radical 62Neotetrazolium (tetrazol purple, TP),

see NTNHI60Nitroblue tetrazolium, see NBTNitro-ditetrazolium salts- "blue granula" 63Nitro group, effect on- fluorescence 46- nitrosensitivity 64- numerical parameters 48, 49- tetrazolium salts 63Nitro-monotetrazolium salts- characteristics 63- for detection of cell injuries 35, 63, 65- dimerization 64- half-wave potential 22- histochemical value 62, 63, 65, 66- localizing qualities 35, 42, 63- and other morphological techniques 65, 66- nitrosensitivity 63, 64- optical characteristics 24- for quantifying 65, 66- reducibilites 35, 42, 48, 63- substantivity 19, 20- TLC 16, 17p-NTTC, 2-p-nitrophenyl-3,5-di(phenyl)-

2H-tetrazolium chloride- UV spectrum 24- visible spectrum 25NT 36, 48, 58, 61- in electron microscopy 44,... formation of mono-formazan 11- and oxidative phosphorylation 50- redox potential 21- reducibility 23, 53, 54, 55, 57- reduction and oxygen 56- spectrum 24- role of substantivity 20- synthesis 11- TLC 17Numerical parameters- and chemical structure 48, 49- employment on histochemica~ properties 49

Subject index . 83

Osmiophilic formazans and tetrazolium salts43, 44, 45, 46

"Osmium black" of BPST 45, 46Osmium tetroxide- reaction with tetrazolium salts 43, 44, 45- postfixation 44Os-TNST 44,45Oxygen- actual redoxpotential 60- and tetrazolinyls 50, 59- effect on tetrazolium reduction 22, 26, 49,

55, 56, 57, 59, 61, 62

Parameters of tetrazolium- formazan system- for histochemical use 18- for histochemical qualification 38,39,40,41,

42,48,49Phenazine methosulfate, PMS 23, 58, 59, 61Phototetrazolinyls 28Photo-TIC 28Phthalhydrazidyl group- as fluorescent group 47- role in BPST 45, 46Polarization optical methods 66

. Polarography 21,22"Poly(nitro-tetrazolium salt)" 22Pulse radiolysis 28

Q-TNBT 45

Redox mediators- PMS 56, 57, 59, 60, 61- mechanism of action 56, 57- Meldola blue 53, 56- methoxy-PMS 56- reaction rates 56, 57- role in tetrazolium reduction 56, 59Redox potentials 21, 22, 52, 53, 60, 61- "actual" potentials 22, 52, 53, 60- definition 22- estimation 21- polarographic dates 22- and reaction rates 22, 57- and respiratory chain 58- of tetrazolium salts 22- variability 22, 52, 53Reducibility of tetrazolium salts 18,21,22,49,

57,59,63- critical value 23

84 . Subject index

- definition 22- and design 22- effectors 22- groups of 52, 53, 55, 56- and Hammett-a-constants 48- and mediators 56, 57- order of precedence 22, 53, 54, 55- and reaction rates 22, 23, 53, 54- and substantivity 23- test systems 22, 23, 52, 53- variability 23Reducing agents for tetrazolium salts 52, 53- ascorbic acid 22, 52, 53- diaphorase 23, 53- glucose-6-phosphate dehydrogenase 53- lactate dehydrogenase 53- NADH-PMS 23- potassium peroxide (K02) 22- succinate dehydrogenase 53- superoxide radical (- O 2) 53, 56, 57, 60, 61, 62- xanthine oxidase 22, 53Respiratory chain- cytochrome-c loosening 60- points of interaction with tetrazolium salts

57,58,59

Scanning transmission electron microscopy 44Substantivity of formazans 18, 19, 43, 63- and chemical structure 38, 39- definition 20- and formazan deposits 20- and histochemical localization 20- and hydrophobic properties 20Substantivity of tetrazolium salts 18,22,43,44,

63- and chemical design 38, 39- definition 19, 20- of flurescent tetrazolium salts 46, 47- histochemical value of 19, 20- influencing factors 19, 20- measuring 19, 20- models for 19, 20- reasons for 19, 20- and tetrazolium reduction 19, 20- and tissue components 19, 20Substitution effects on formazans 29, 38, 39,

40,41- N-N'-dimerization 37- NOrinsertion 37, 42

- red formazans 35, 36, 37- spectra 31Substitution effects on tetrazolium salts 36 et

seq., 42Succinate dehydrogenase- test system for histochemical suitability 49,

50,53,56Superoxide dismutase- role in tetrazolium reduction 57, 59, 60, 61,

62Superoxide radical (.02)

- as tetrazolium reducing agent 53, 57, 59, 60,61

Structure-staining-correlations 36, 48, 49

Tt 36TC-NBT 44Tetranitroblue tetrazolium, TNBT 37, 42, 44,

52,58,62- in electron microscopy 44- half-wave potential 22- localizing qualities 35, 62, 63- reducibility 23, 53, 54, 55- reduction and oxygen 35, 56, 62, 63- synthesis 11- TLC 17- UV spectrum 24TNBT formazan- "nitrosensitivity" 64- spectrum of "aci-form" 33- TLC 17- visible spectrum 25, 27TNBT1h- "nitrosensitivity" 64- reductibility 23Tetrazoles 5, 51, 62Tetrazolinyls- absorption characteristics 26, 52- ditetrazolinyls 51- formation of 51, 52, 60- formating formazans 28, 51, 62- forming tetrazoles 62- stability 51, 52, 62- in tetrazolium reduction 31, 52, 59, 65Tetrazolium-formazan system- biological uses of 49, 50Tetrazolium methods- for dehydrogenases 50- for hydrolases 54

Tetrazolium reduction- aerobic 59, 60, 61, 62- anaerobic 59, 61- and reducibility 54, 55, 56, 61, 62- factors affecting 22, 23, 50, 55, 61, 62- by flavins 59, 60, 61- by flavin-like models 59, 60, 61- by heme-containing compounds 60- manometrically 53- mechanism 52, 57, 58, 59, 60, 61, 62- models of 61,62- non-enzymatic 19,53, 54- non-reversibility 21, 22- polarographically 53- and respiratory chain 58, 59- role of detergents 57, 61, 62- role of macromolecules 55, 56- role of mediators 56, 61, 62- role of oxygen 23, 55, 59, 61, 62- role of PMS 61, 62- role of radical intermediates 61, 62- by substrates and enzymes 60- tetrazolium concentration 55, 56, 61, 62- velocities 531-H-Tetrazolium salts 22-H-Tetrazolium salts 2- analytical chemistry of 2, 15, 16- in analytical chemistry 1, 2, 14, 17, 18- betaines 4, 5- biological actions 50- in biological sciences I, 14, 15- contaminations in 15- dimerization 36, 62- effect on enzymes 50- for electron microscopy 43, 44, 45, 46- electrophoresis 17- elementary analysis 15- extinction coefficients 24- fluorescent 46, 47- half-wave potentials 22- Hammett-(J-constants 48, 49- history 1- IR-spectroscopy 17- isomeric forms 2, 3, 4, 16- light stable 28- melting points 14, 15- numbering system 2, 3, 4, 5, 6- optical characteristics 24 et seq.- "optimal salt" 17, 18

Subject index . 85

- osmiophilic salts 43, 44, 45- "oxygen-insensitive" 23, 55, 56, 57- "oxygen-sensitive" 23, 55, 57, 59- parameters of histochemical value 18, 19- penetration into sections 63- "pick-up-order" 57, 58- photodehydrogenation 28, 29- properties 3, 4, 14, 15, 18- properties for histochemistry 17, 18- purification methods 14- purity of IS, 16, 36, 37- qualification for histochemistry 35- radicals of, see tetrazolinyls- redox potentials 57, 58- reducibilites 18, 21, 22, 23- reduction of 4,21,52, 57, 58- and respiratory chain 57, 58- as resonance hybrids 4- sensitivity to light 14, 15, 28- structure 2, 3, 4, 8, 9- structure and histochemistry 1, 3, 17- substantivity 18- substituents 3, 4, 42- synthesis 2, 10, 11, 12, 13- TLC 15, 16, 17- types of 4, 5, 36, 42m-3-TNTIC- "nitrosensitivity" 64- reducibility 23m-5-TNTIC- "nitrosensitivity" 64- reducibility 23p-TNTTC salt 37- "nitrosensitivity" 64- reducibility 23, 53- UV spectrum 24p-TNITC formazan- spectrum of "aci-form" 33- visible spectrum 251,3,5-Tri(phenyl)-l-methyl-formazan 27Triphenyltetrazolium chloride TIC 18,35,48,

58,61- effect of light 28- half-wave potential 22- histochemical quality 36, 37- and oxidative phosphorylation 50- radical 26- redox potential 21, 22- reducibility 23, 35, 36, 53, 55, 57

86 . Subject index

- TLC 17- UV spectrum 24TIC formazan- isomeric forms 29, 30- localizing quality 35, 36- TLC 17- visible spectrum 25, 27Tetrazolium violet lV- localizing qualities 35- reducibility 23, 35, 53, 55- TLC 17

lV formazan- isomeric forms 27- visible spectrum 25

Ubiquinone 58, 60

Xanthine oxidase- test for tetrazolium reducibility 23

YT44