(195) Pella, E., Mikrochim. Acta, 1968,13. (196) Ibid., 1969, 490. (197) Pennington, S., hleloan, C. E., ANAL.CHEM.,39, 119 (1967). (198) Perez-Bustamante, J. A., Burrielhlarti, F., Inform. Quim. Anal. Pura Apl. Ind., 22, 25 (1968). (199) Perkin-Elmer Corp., Norwalk, Conn., Bull., Model 240 Elemental Analyzer, 1969. (200) Pitre, D., Grandi, M.,Xikrochim. Acta. 1967. 347. (201) Pietrogrande, A., Dalla Fini, G., ibid., p 1168. (202) Zbid., 1968, 228. (203) Podorozhanskii, 11. 1I., Zeidlits, E. M., Eru, I. I., Zavod. Lab., 33, 697 i1967). (204) Prokopov, T. S., Mikrochim. Acta, 1968, 675. (205) Raspanti, G., 2. Anal. Chem., 225, 24 (1967). (206) Rauch, P., Tykva, R., Chem. Listy, 61, 1669 (1967). (207) Reznitskaya, T. V.,Burtseva, E. I., Zh. Anal. Khim. 21, 1132 (1966). (208) Rison, 11. H., Barber, W. H., Wilkniss, P., ANAL. CHEM., 39, 1028 (1967). (209) Rudran. K.. Kamath. P. R.. Microchem. J . . 11: 481 119661. ’ (210) Saharoiici, R., Pascal, P., Rev. Chim. (Bucharest) 17, 560 (1966). (211) Saran, J., Khanna, P. N., Banerji, S., Zaidi, S. B. N.. Nzkrochim. Acta. 1968, 1124. (212) Sato, T., Takahashi, T., Ohkoshi, S., Japan Analyst, 16, 309 (1967). (213) Scheidl, F., Microchem. J . , 13, 155 (1968). (214) Schlunz, AI., Koster-Pflugmacher, A., 2. Anal. Chem., 232, 93 (1967). (215) Schoniger, W., Mzcrochem. J., 11, 469 (1966). 1216) Schultz. R. B. T.. Z . Lebensm.Cnters. Fokch., 134, 353 (1967). (217) Schwarz-Bergkampf, E., Mikrochim. Acta, 1967, 1001;
(218) Scott, B. F., Kennally, J. R., ANAL. CHEM.,38, 1404 (1966). (219) Scroggins, L. H., Microchem. J . , 13, 385 (1968). (220) Selig, W., Z . Anal. Chem., 234, 261 (1968). (221) Sels, F., Demon, P., Mikrochim. Acta, 1969, 530. 12221 Shanina. T. RI.. Gel’man. K. E.. Mikhailovskaya, V.‘ S., Zh. Anal: Khim., 22, 782 (1967). 1223) Shibazaki, T., Koibuchi, hI., J . Pharm. Soc. Japan, 88, 140 (1965). (224) Sloane-Stanley, G. H., Biochem. J., 104, 293 (1967). (225) Smith, A. J., Cooper, F. F. Jr., Rice, J. O., Shaner, W. C., Jr., Anal. Chim. Acta, 40, 341 (1968). (226) Smoczkiewiczowa, A., Augustyniak, J., hleissner, W., Chem. Anal. (Warsaw), 12, 629 (1967). 1227) Sro. L.. Chem. Prum.. 17. 390 (1967): 12281 Stanfer. C. H.. Dworkin. R. D.. A ~ A LCHI&., . 40, is91 (1968j. (229) Starcuk, Z., Cupak, LI., Chem. Listy, 60, 1543 (1966). (230) Strukova, hl. P., Fedorova, G. A., Zh. Anal. Khzm.. 21. 509 11966). (231) Strukova, Ai. P:, Kashiricheva, I. I., Lapshova, .4. A., ibid., 22, 1110 (1967). (232) Strukova, hl. P., Kirillova, T. V., ibid., 21, 1236 (1966). (233) Strukova, hl. P., Kotova, V. N., ibid., 22, 1239 (1967). (234) Taves, D. R., ANAL. CHEM.,40, 204 (1968). (235) Technicon Corp., Tarrytown, N. Y., Bull. CHNO Analyzer, 1969. (236) Terent’eva, A. P., Bondarevskaya, N. A,, Gradskova, N. A,, Kropotova, E. D., Zh. Anal. Khim., 22, 454 (1967). (237) Terent’eva, A. P., Volodina, AI. A., Fursova, E. G., i b i d . , p 640. (238) Terent,’eva, E. A., Bernatskaya, 31. V., ibid., 21, 870 (1966). ~
(239) Terent’eva, E. A., Smirnova, N. N., Zavod. Lab., 32, 924 (1966). (240) Terent’eva, E. A., Vinogradova, E. N., Akimov, N. P., ibid., 34, 414
__
(1968\. ~ - -
(241) Thomas, A. H., Philadelphia, Pa., Bull., hlodel 35 hlicro C-H Analyzer, 1969. (242) Truffert, L., Favert, M.,Le Gall, Y., Ann. Falsij. Expert. Chim., 60, 27.5 11967). (243) Trutnovsky, H., Mikrochzm. Acta, 1968, 97. (244) Trutnovsky, H., Z . Anal. Chem., 232, 116 (1967). (245) Turuta, A., Crisan, T., Pal, M., Reo. Chim., 18, 306 (1967). (246) Tykva, R., Collect. Czech. Chem. Commun., 32, 2001 (1967). (247) Tykva, R., Int. J . Appl. Radzat. Isotopes, 18, 45 (1967). (248) Uhle, L., 2. -4nal. Chem., 231, 194 (1967). (249) Utsumi. S.. hlachida. W.., Ito., S.., Japan Anaiyst,’l6, 674 (1967). (250) Voegeli, P., Christen, F., Z . Anal. Chem., 233, 175 (1968). (251) Walisch, W., Humme, G., Mzkrochtm. Acta, 1968, 748. (252) Walisch, R., Jaenicke, O., ibid., 1967, 1147. (253) Walisch, W., Xarks, W., ibid., p. 10.51
(2;4YWalisch, W., Schafer, K., ibid., 1968, 765. (255) Wheeler, P. P., Fauth, M. I., AKAL.CHEM.,38, 1970 (1966). (256) Wineburg, J. P., ibid., 40, 1744 (1968). (257) -%right, H., Explosirsto$e, 14, 274 (1966). (258) Wronski, hl., Bald, E., Chem. Anal. (Warsaw), 12, 863 (1967). (259) Yamazaki, 11.) Ishihama, H., Kasida, Y., Znt. J . A p p l . Radiat. Isotopes, 17, 134 (1966). (260) Yeh, C. S., Microchem. J . , 14, 279 (1969).
Chemical Microscopy G. Cocks,
George
T(W
Cornell University, lfhaca, N . Y.
iii this series covered the two-year period from January 1965 through December 1967. Some earlier articles and books omitted from previous reviews are included in this review. I t is the purpose of this review to report on articles and books of potential interest to those mho use the microscope to solve chemical problems. However, because “chemical problems” are so diverse, and because the microscope is used in so many fields, it was necessary to restrict this review in several ways. KO attempt has been made to report publications in the fields of biochemistry, petrography, or metallography unless they were deemed to be of direct interest to cheniists or chemical engineers. The chemical applications of electron microscopy are the subject of a separate review, and they are not reHE PREVIOUS RCVILW
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port’ed here. References to publications in the fields of optics and crystallography are included if they appear to be of interest to chemical microscopists. Past reviews in this series have included a section on meetings and symposia. Because meetings and symposia have become so numerous, and because of the desirability of reducing length of this review, reports of meetings and symposia have been omitted. Printed proceedings of meetings or published articles based on papers presented a t meetings are reported. The great’ diversity and number of publications of possible interest’ to chemical microscopists makes it impossible to find and review all of them. Therefore the author would appreciate comments or suggestions, particularly with regard to the omission of important published art’icles.
ANALYTICAL CHEMISTRY, VOL. 42, NO. 5, APRIL 1970
BOOKS OF GENERAL INTEREST
A book entitled “The Microscope, A Practical Guide” (1%) has been written by ?Seedham. This book and a third edition of Barer’s excellent “Lecture Notes on the Use of t,he Xicroscope” (9) proyide an introduction to microscopy for beginners. Barer’i book has been expanded by including appendices on phase-cont’rast microscopy, and on the use of research type illuminators. Fraii~oiiJs “Progress in Microscopy” has been translat’ed into German by Ludwig (54). The translator has also included revisions of the sections on phase and interference microscopy. Two more books iii the series entitled “hdvances in Optical and Electron llicroscopy” ( I f ) have appeared. Volume 2 contains four chapters of direct interest to light microscopists.
I n t’he first chapter, G. T. Reynolds describes light intensification systems as applied to light microscopy and ho the detection of radioactive t,racers. Intensifier film systems are analyzed and bhe statisbics of the multiplication process are discussed. Welford’s chapicroter on the “AIach Effect and the I11‘ scope” is a state-of-the-art report discussing errors which may affect the microscopical determination of the locat’ion of the edge of a n object. Errors arising from physical phenomena are dealt with as well as t h e visual errors attributable to the Mach effect. T h e review entitled “Digital Transformation and Computer Analysis of Microscopic Images” is a description of work carried out jointly by groups from the University of Pennsylvania and from the Airborne Instruments laboratory. The work described is directed toward the development of instruments and techniques for the automatic interpretation of microscopical images, for example, the clinical diagnosis of white blood cells and human chromosomes. K o r k being done in other laborat,oriesis also reviewed. There is also a brief chapter on “;iutoradiography and the Photographic Process” in which the factors affecting the preparation of autoradiographs for light or electron microscol)y are discussed, Other chapters in the book are concerned 1%-ith high voltage electron microscopy, with the fixation and embedding of tissues, anti with field ion microscopy. Volume 3 of “aidvances in Opt,icaland Electron Microscopy” (22) contains two chapters of direct interest to light microscopists. “Zoom Systems in Alicroscopy” presents the historical background for t h e development of these systems. It also discusses the design of zoom systems, describes some actual zoom microscopes7 and finally discusses trends and limitatioiis in the design of zoom systems. .I second chapter is devoted to “Mensuration Methods in Optical Xicroscopy.” This is a review of the apparatu.q, the techniques, and t h e theory of microscopical mensuration. Fully automatic methods are not discussed. The measurement of dimeiisioiis in the image plane and perpendicular to it are covered. lleasurement of areas, volumes, and angles are also coiisidered. “The Particle - 4 t h ” ( l o b ) by N c Crone, Draftz, and Delly contains a large number of excellent color micrographs of part’icles. To aid in identification, a binary classification is presented along \vith an extensive discussion of microscopical techniques for part,icle identification. There is also a section on dispersion staining and an extensive bibliography containing 1185 references
on various aspects of microscopy. The same authors published a series of pamphlets called “The Particle Analyst” (106) in which they anslvered quest,ions pertaining to microscopy and discussed many topics of interest to microscopists. These pamphlets came out semi-monthly during 1968. Publication has been discontinued, but’ the compiled 1968 set is still available from the publisher. Books on optical crystallography include new editions of Hartshoriie and Stuart’s “Practical Optical Crystallography” (74) and JJ-ahlstroni’s “Optical Crystallography” (202). The former book is a n abridgement of the author’s more extensive “Crystals and the Polarizing Microscope.” T h e most significant, change in Kahlstrom’s book is the inclusion of references follon-in$ most chapters; however, the sections on eubstage illuminators aiid optical activity have been extended a i d a chapter on dispersion st’aiiiiiig has been added. Bishop has published a new book entitled “=in Outline of Crystal Morphology” (21). Zussnian’s book “Physical Methods in Determinative llineralogy” (214) includes a section on optical microscopy using 110th transmitted and reflected light. Kley’s classic “Organische JIikrochemische AInalyse”has been translated into English by R. E. Stevens (54). T h e techniques for the microscopical identification of organic compounds described deserve to be more widely applied by organic chemists. Tolansky has published trvo new books on interferometry, “Interference Xicroscopy for the Biologist” (192) and “hlicrostructures of Surfaces Using Interferometry” (191). The latter is a n introduction to the techniques of interferometry, and t h e former contains many suggestions on how the biologist can make use of interferometry. “Medical Colour I’hotoinicrography” (185)by Szabo is a guide for physicians, biologists, and others for n-hom color photomicrography is either part of their routine work or is needed in special research projects. I t ContainL; much practical information for anyoiic interested in color 1)hotomicrogral)hy. Stevens has r e v i d xiid greatly enlarged his book on “Alicrophotography: Photography and Photofabrication a t Extreme Resolution” (185). This book contains much information of direct interest to microscopists. ARTICLES OF GENERAL INTEREST
An article by Uradbury gives a brief history of the development of the mechaiiical and optical parts of the micro. LlcCroiie and Hertrich (107) describe a home-built laminar flow clean bench which easily fulfills the coiiditioiis for :i class 10,000 clean room. Delly (38, 106) diqcusses particle reference staiidardq, i.e., sets of known particles which are helpful for the identification of uiikno~vnparticulates. Sources for thrse sets of st’andards are given. The microscol)e is potentially an important tool for exploring estraterrestrial bodies. .I niicrosco1)e suitable for soft-landing rockets is difficult to build particularly lircausc of the problems of prclmring s1jrcimens and manipulating thP microscope. Soffeii (1S1) discusses three instrumcnts which h a w heen suggrsted for this piirpo>c. Chemical microsco1)ists arc often faced with the necessity for workiiig with hydrofluoric a d . Richardson (258) suggests that damage to metallograph objectives can be eliminated by treating the specimen with ammonium pentaborate after it has been exposed to
HF .
OPTICS
~iprimary coiiccrn of the microicopist is the resolution ohtained with his microscop. Orhaug (130) writes on the resolution of imaging systems nntl defines angular rcsolut ioii as the ability
ANALYTICAL CHEMISTRY, VOL. 42, NO. 5, APRIL 1970
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of a system to estimate the position of a point' source object observed in a background of gaussian noise. H e applies the theory of statistical estimation to this situat'ion. As mentioned above Zieler (212) gives a good discussion of resolving power. Jones (86) describes two apparent exceptions to -4bbB's theory of resolution; namely, resolution in red light and not' in blue light when annular bright-field illuminat,ion is used, aiid t h e case where a diffracting object is illuminated by diffracted beams from a n underlying diffracting object,. Hewlett (76) also describes a case where visibility and resolution of a periodic structure in oblique dark-field illumination seemed to be anomalous. Both this case and the cases described by Jones were compatible with AbbB's theory correctly applied. Hewlett concludes that the general statement t h a t resolution with dark-field is equal to t h a t with bright-field illumination is not true, Martin (115) describes a method of improving the resolving power of a glass lens by means of a Hoppe zoneplate. Payne (134) discusses bright-field illumination for microscopy describing the types of illumination, the basic requirements for good illumination, and the types of illuminators and lamps available. Reference (12) contains a chapter by Benford and Rosenberger on zoom syst'ems in microscopy. Martin (116) reviews methods of obtaining stereoscopic images using single-objective binocular microscopes, and introduces a new method using semitransparent half shade caps over the eyepieces. ,4n analysis of stereoscopic imagery in single objective stereoscopic microscopes is given by Kavanagh (91). The light microscope as a n optical diffractometer is the subject of a paper by Gall (62). Uhlig (199) gives a brief simple explanation of how anti-reflection coatings on optical surfaces work. H e also tells how such coatings are utilized in the design and manufacture of microscope lenses. Llart'in (117) describes the use of phase plates located at t'he Ramsden disk of the eyepiece. This location is convenient for converting any objective into a phase objective. It is especially useful for objectives whose back focal planes lie within t'he lens itself. Polarized light can be analyzed using a stereographic projection of the Poincark sphere (113). R o o d and Mills (206) give a proof that trirefringence does not occur in the region of linear optical effects. Spry et al. (182) discuss the optical phenomena associated with Brazil-twin boundaries in quartz, and the optical activity occurring in nonenant'iomorphous AgGaS is discussed by Hobden (77). The AgGaS crystal is interesting in that optical rot'ation can be right', left, or zero depending on the 116 R
direction in which light t'ravels through the crystal. R a t h and Pohl (152, 153) discuss the effect of lens surfaces and the first order red plate on interference figures. Goldstein (67) presents two new methods of detect'ing dichroism. They also discuss the theory of dichroism and t'he relat'ionship of dichroism to the color of stained histological specimens. The influence of focusing errors on the measurement of phase differences in biological objects using polarized-light microscopy is demonstrated by Hutschenreiter and Schemer (82),and Thetford and Simmens (190) explain how the interference pattern of a cylindrical fiber is displaced depending on the refractive index of the medium in which the fiber is mounted. Haydon (75) suggests that one source of image contrast in electron microscopical images appears to be analogous to the contrast' obtained in light microscopy when the object is mounted in a material of widely different refractive index. As a result, electron stains on t'he surface of a specimen are more effective than those within a specimen. Edholm (42) has made a study of the effect of lens aperture on the energy distribution in the image of a nonrefracting sphere. A number of papers are concerned with errors in microscopical measurements caused by optical effects. Among these are a n experimental study of the effects of the refractive index of t'he mount'ing medium on the apparent diameter of latex part'icles and glass fibers (177), the effect of the numerical aperture of the microscope objective on the determination of film t'hickness using interferometric methods (139), and errors in a real analyses resulting from the thickness of the specimen (66). Galjaard (61) discusses the sources of error in the determination of the drymass concentration in tissue sections considering the measurement of section thickness, the effect of the immersion medium, the type of microscope used, and the method of measuring the optical path difference. Rowe (165) suggests a precise method of focusing a microscope when using coherent (laser) illumination. The method makes use of a reference beam projected onto the specimen. This reference beam interferes wit,h the illuminating beam to form fringes which can be used for precise focusing. The Mach effect, as was noted earlier, was the subject of a review by Welford (11). Rowe has published two articles dealing with t'his effect (166, 167). I n one he finds that small focusing errors affect the posit,ion of the Mach bands, and in the other he shows that there is little difference in the Mach effect as observed using monocular or binocular microscopes.
ANALYTICAL CHEMISTRY, VOL. 42, NO. 5, APRIL 1970
INSTRUMENTS AND EQUIPMENT
Microscopes, Objectives, and Eyepieces. Reynolds (11, 166) has described a' microscope, fitted with a sensitive image intensifier unit, capable of observing directly t,he location of radioactive tracers in a specimen. A simple microfluorimeter is described by Stoward (187), aiid Barer (10) tells how t o obtain the maximum intensity of fluorescence by illuminating t,he specimen using both incident and transmitted illuminat'ors. Pearse describes a microspectrofluorimeter with epi-illumination and photon counting (155). This instrument is a modification of t'he Leitz microspectrograph. Pet& et al. (158) have invented a "tandem scanning reflected-light microscope" and this instrument was modified and improved by Davidovits and Egger (35). It operates by scanning a beam of laser light across the specimen, and using the reflected light to modulate the beam of a synchronously scanned oscilloscope. Through the use of polarizing filters and pinholes, it is possible to exclude stray light, to such an extent' that images of buried layers of living tissues can be formed. Bennett (19) describes improvements which he has made on a single beam reflectance photometer. H e used this instrument to measure the reflectance of polished mineral grains. A more sophisticated photometer for reflected light microscopy has been described by Larsoii (9'7), It is a double beam instrument with a null balancing photomet'er covering a wavelength range from 400 to 700 nm. It has a mechanically driven stage and a n x-y recorder and was designed for the quantitat'ive photometric measurement of reflectivit'y by polarized light microscopy. Jones et al. (88) report on a n apparatus for reflectivity measurements of reactive and radioactive materials, The photometer was assembled from commercially available components. The microscope was built into a glove box. h device, manufact'ured by Zeiss Jena, for producing phase-cont'rast like or shearing interference microscopical images a t will has been described by Beyer (20). The device is called Interphako. Hollis and Kelly (80) have described a vertical illuminator microscope for multiple beam interferometry. A photoelectric phase-measuring microscope has been reported by Smith (178). A continuously driven compeasat'or causes sinusoidal fluctuation in the intensity of the transmitted light. When a n object is placed on the stage, the phase relationship is displaced in direct, proportion to the retardation. This signal is picked up b y a photomukiplier and transformed elect'ronically to give a direct measurement of phase shift.
Martin has written two articles describing special eyepiece caps. One of these (117) deals with a phase plate which, when mounted at the Ramsden disk, transforms the microscope into a phase microscope. The second article (116) describes caps which are used to obtain stereoscopic images with monoobject’ive microscopes. A new design of reflecting objective consisting of two concentric spherical mirrors has been reported b y Beck ( 1 7 ) . T h e author claims t h a t this design has less obscuring of the aperture and higher resolution than ordinary reflecting objectives. Work on the development of inst’ruments and techniques for the automatic interpretation of microscopical images has been ment’ioned above (11). Some further description of these microscopes may be found below under Automatic Image hnalysis. Stages. T h e applications of h o t st’age microscopy are listed a n d discussed b y 1fcCrone (104). Hamer (72) has given information on how to build a constant t’emperature controller for t h e Kofler hot stage. Loasby (101) describes a low inertia hot-cold microscope stage. It is cooled by gaseous nitrogen (or other gas) and is heated electrically. It covers a range from -200 to +200 “ C and maintains temperatures to i l “C. It provides for keeping the specimen under vacuum and is designed for use on a n inverted t,ype of microscope. Price (147) describes an air curtain incubator which he uses for time-lapse cinemicroscopy. Temperature regulation is k0.5 “C. A simple, hot wire, microforge used to shape glass tools for micromanipulators is described by Cook (32) who also gives some instruction in its use. l l o r e specialized stages include a pressurized specimen chamber for light microscopy capable of containing pressures u p to 10 at,mospheres (16) and a n electron bombardment8 luminescence stage for optical microscopy (73). Miscellaneous. X number of exposure meters for photomicrography have been described. Brown’s photometer (2b) consists of a simple photoresist’ive element mounted on a probe so that it can be positioned where desired in the image plane. Saurer et al. make use of scintillation counting equipment in their sensitive photometer (169). A simple automatic exposure timer is described by Ruzicka (168), and Yos (209) reports on photomicrography with the Science and M e c h a n i c s exposure meter. A simplified time lapse cinemicrographic unit which drives t,he camera continuously and uses a synchronized clectronic fla,ph for exposure is described by Burton (26). Robb and Jabs (160) also make use of electronic flash for their cinephotomicrographic apparatus.
A photoelectric apparatus which measures refractive index b y sensing a phase contrast image while a n index liquid is being heated has been described b y Chromy (29). A similar apparatus has been used b y Vaughan (201) to observe and record the melting points of single crystals. Vaughan uses a polarizing microscope instead of a phase microscope. hlagill and Gunning (111) have built a simple microtome for cutting sections of plastic embedded materials as thin as 1 pm. This instrument can be built in a n ordinary machine shop. POLARIZED LIGHT MICROSCOPY
A number of publications relat’ing to polarized light microscopy have been reported elsewhere in this review under Books of General Interest (21, 74, 202); under Optics (66, 67, 82, 113, 152, 153, 182, 190, 206); under 1nstrumeiit.s (35, 97, 178, $01); and under Infrared and Ultraviolet Light Microscopy (204). A book b y Zussman (214) on “Physical Methods in Determinative Mineralogy” includes sections on microscopy by transmitted and reflected light. T h e applications of polarizing microscopy techniques to the study of the microstruct,ure of glass reinforced plastics are described b y F r y et al. ( 5 7 ) . Nicholson (228) has suggested a technique for estimating the thickness of a thin section by embedding sand around the specimen and measuring t h e retardation of t h e sectioned sand grains showing maximum birefringence. I n discussing Nicholson’s technique, Ehlers (43) point’s out t h a t t h e method can be improved by substituting gypsum for the sand grains. Gypsum has (0011 cleavage and lies on the cleavage plane, thus orienting itself and making measurement of the section thickness easier. Nettelnstroth (127) reports on the identification of synthetic fibers using measurements of specific birefringence. Lieder (99) reports on the curious morzinc selenite phology of anhydrous (ZnSeO,) which crystallizes as a double wedge. This material is useful for demonstrating the interference colors.
a
INFRARED AND ULTRAVIOLET LIGHT MICROSCOPY
Casida (27) has used infrared color photomicrography to reveal bacteria selectively. Yoder (208) discusses temperature measurement with a n infrared microscope. The temperature of object,s, approaching in size the wavelength of the radiation, can be measured. It is also possible to obtain thermographic images of microstructures. An ultraviolet microspectrophotometer for measurements with polarized light has been described by Wetzel et al. (204).
PHASE CONTRAST MICROSCOPY
Pluta has published five art’icles dealing with equipment for phase contrast microscopy; a n apparatus for phase contrast using positive phase rings made of soot and a dielectric substance (I@), stereoscopic phase contrast microscope (141), properties of the amplitudecont,rast microscope with soot amplitude rings (l&), a phase-contrast device with positive and negative image cont.rast (143), and a highly sensitive phase contrast device (145). RIondal (122) has examined the effect’s of varying the conjugate area of the phase plate in a phase contrast system. H e has also studied phase contrast microscopy in part’ially coherent’ light (121). Martin (117) describes phase plates which are inserted in the eyepoint of the microscope. These plates make it possible t’o produce phase contrast illumination with any objective. INTERFERENCE MICROSCOPY
Tolansky has published two books; one dealing with the biological applications of interference microscopy (152) and one dealing with the “Rficrostructure of Surfaces Csing Interferometry” (151). H e has also published a n article entitled “Multiple-Beam Interferometry in Analytical Metallography” (153). Elsewhere in this review under t h e heading Instruments t’hree articles (20, 80, 178) dealing u 4 h interference microscopy have been reported. T h e theoretical background for a type of common p a t h interferometer, exemplified by the axial twist and planar inversion interferometers, is discussed by Alpiar ( 4 ) . H e describes t h e devices and their potent,ial application to interference microscopy. 9 doubly refracting interference microscope with variable image duplicat,ion and half-shade eyepieces is described by Pluta (144). I n a long article containing numerous micrographs of test objects of known geometry, Padawer (131) presents a n experimental basis for the int,erpret’ation of images formed by the Xomarski interferencecontrast microscope. Arregger ( 7 ) discusses the application of reflected light interference microscopy to the solution of technical problems. Snow and Vandewarker (180) describe a two beam interference microscope in which a hologram is used to generate one of the beams. This is a modified bright-field microscope capable of producing horizontal, or oblique section int,erferograms, differential or totally doubled interferograms in real or recorded time. Magill and Wilson (212) also discuss applications of a holographic interference microscope. Simniens (176) describes refractive index measuremeiit,s on birefringent materials using interference microscopy. The method is a double immersion
ANALYTICAL CHEMISTRY, VOL. 42, NO. 5, APRIL 1970
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method suitable for samples whose thickness is not known. Neugebauer and Fragstein (125) present methods of measuring both thickness and refractive index of phase objects using a polarizing interference microscope. One method is as accurate as Tolansky’s interferometric methods. Pliskin and Esch (139) discuss the effect of the numerical aperture of microscope objectives on film thickness determination by interferometric methods. Special applications of interference transmission microscopy to textile fiber problems are suggested by Poling and Hollies (146). McKee and Woods (110) describe a n accurate method of measuring the birefringence and refractive indices of textile fibers. T h e method involves channelled spectra from the fibers in the Baker interference microscope.
WAVEFRONT RECONSTRUCTION MICROSCOPY
A review of the state of the a r t of holography, including holographic (wavefront reconstruction) microscopy has been edited by Kallard (89). Gabor has written an article entitled “The Outlook for Holography” (GO), and Rogers and Palmer (163) discuss the possibilities of X-ray holographic microscopy. A method of reconstructing a three dimensional image of a microscopic sample using holographic techniques is described by Toth and Collins (195). I n this method a lens is placed between the object and the hologram plate to increase the effective numerical aperture of the system. Anderson ( 6 ) discusses carrier suppression and restoration in holographlc microscopy. This is a method of obtaining variable dark-field and phase contrast images. Hohberg (79) proposes a method of improving the resolving power of a microscopic system with a limited entrance pupil by enlargement of the effective pupil by holographic means. Champagne and Massey (98) discuss the factors which limit the numerical apcrture (and hence resolving power) in holograms. Hologram techniques for particle size analyses, the theory behind the techniques and the equipment used for preparing holograms of moving particles are described by Zinky ($13). van Ligten (200) reviews holographic microscopy and compares it with conventional light microscopy. Fourney et al. (53) describe a method for determining the velocity and size of aerosol particles using holography. Holographic techniques have also been used to observe crystal growth from the melt (109). 118R
FLUORESCENCE MICROSCOPY
Only a few references to fluorescence microscopy have been selected for inclusion in this review. Most of the work in fluorescence microscopy has been done by biologists and biochemists. A few references to fluorescence microscopy can be found in this review under the headings Instruments (10, 11, 135, 156,187) and Photomicrography (210).
anomalous resolution of diatom structures which consist of rectangular or triangular arrays. Abb6’s formula, which was developed for gratings, must be modified for application to these structures because additional diffraction maxima occur. The authors investigate some claims for dark-field resolution of diatoms. TECHNIQUES FOR SPECIMEN PREPARATION
SCHLIEREN MICROSCOPY
Dodd (41) describes a schlieren microscope and discusses the purposes of the various kinds of uses to which it can be put. Among the applications are the study of concentration gradients, heat gradients, and the observation of biological materials. MICROPHOTOMETRY AND MICROSPECTROPHOTOMETRY
References to articles describing microphotometers for reflected light can be found under Inst’ruments (19,88, 97). These instruments have been used primarily for examination of ores. I n order to adjust and standardize t’hese instruments, reflectivity st’andardsare needed. Nayak, in an article entitled “The Status of Reflectivity Standards in Ore hlicroscopy” (124), reviews the substances used as reflectivity standards and gives the most recently determined reflectivities of a number of them. Three microspect’rophotometers have been reported in the recent literature. Tonna and Rogers (194) describe an instrument incorporating a fixed double aperture optical system and an electronically controlled automatic scanning microscope stage. They report on t’he use of this instrument’ for photodensitometry and microspectrophotometry of cells using continuous automatic tlvo dimensional scanning mode of operation. Engle and Freed (46)describe a “doublebeam vibrating mirror flying-spot scanning-integrating microspectrophotometer” which thejr have used to study living tissue. Wetzel et al. (104) describe a n ultraviolet microspectrophotometer for measurements with polarized light. This is a double-beam instrument, with automatie balancing and recording. It also allows the use of an ultraviolet sensitive image converter and photographic recording of the image. h b sorption spectra for objects of the order of 1 fim in diameter can be made in t h e wavelength range from 240 to 360 nm. With t’he aid of a compensator, dichroism measurements of small objects can be made. DARK-FIELD AND ULTRAMICROSCOPY
Hewlett (76) has investigated the “visibility and resolution of periodic structures in oblique dark-ground illumination.” H e discusses the apparently
ANALYTICAL CHEMISTRY, VOL. 42, NO. 5, APRIL 1970
Mounting a n d Embedding. M o u n t ing media for particle identification are described a n d their use is discussed by Delly (39). Schrader and Weber (17‘3) have investigated the applicability of various media for mounting thin sections of polymers. ,I prime consideration is that’ the media d o not affect the specimen appreciably. The sectioning of friable deposits of minerals or other materials presents a problem in that bhey must be embedded without disturbing them. Zeidler and Ramsden (211) spray such deposit’swith poly(vinylforma1) solut’ionseveral times, then embed the consolidat’ed mass in cold setting polyester resin. Microtomy. G r a h a m (69) reports o n tracer diffusion studies of metals using a microtome. He has cut 2 pm sections of Zr, Ti, Fe, Pd, h g , and Cu using a Jung JIet’al X c r o t o m e and a tungsten carbide knife. Although he used the secbions primarily for tracer work, interferograms of the cut surfaces were prepared. X simplified technique for sharpening microtome knives has been worked out by Quackenbush (149). He replaces the usual spring back with ball bearings, and sharpens using grits on a plate glass surface. Grinding a n d Polishing. G r a y (70) has published a n article on metallographj- entit,led “Xew Methods Point the K a y (Netallography).” Freuiid (56) has edited a nelv book, Band 111, Teil I, of the “Handbuch der IIikroskopie”), which deals with the preparation of metallogrxphic specimens. This book contains one of the few discussions of the theory of etching, especially color etching. I t also has sections on surface replication and hot stage microscopy. Moreland (123) describes techniques for the preparst,ion of polished thin sections of minerals to be used for reflection and transmission microscopy and electron probe analysis. H e uses three stages of polishing on diamond and alumina grits. Staining a n d Etching. S t a u b and McCall (185) have increased the microscopical contrast of phases having similar reflectivities by coating the meta~llogra~~hically prepared specimen with a thin layer of Parlodion. A 0.75% solution of Parlodion is flowed over the surface of the specimen and allowed t o dry. hpparently this layer produces
interference effects which depend on t,he depth of etch. Pate1 and Koshy (133) have etched synthetic and naturalBaS04 single crystals on their (001 1 and { 1101 cleavage faces. The resulting etch pits reveal edge dislocations only on synthetic B a s 0 4 and both edge and screw dislocations on natural BaSO4. Bender (18) has used a three-step technique involving electroetching, microhardness test’ing, and preferential etching for the qualitat’ive analysis of uranium-plutonium mixed carbide nuclear fuels. Miscellaneous M e t h o d s . T h e isolation of radioactive fallout particles for microscopical examination is a difficult task. Svihla (188) has isolated such particles using successive dilutions of t’he particles, keeping track of t h e radioactive ones by autoradiography. Adams and Tong ( 2 ) have used a microscope-laser system as a microsampling device for refract.ory materials. Samples of the order of 10-8 gram can be separated by placing a coverglass over t h e spot to be sampled and then vaporizing the selected spot wit,h a laser beam. Sampling can be done in a vacuum system if that is desirable. Page (132) has developed a method for determining t’he fibrillar angle in wood tracheids. The lumens of the tracheids are filled with mercury using a Porosimeter. If t h e specimens are then observed in reflected polarized light, only one wall of the tracheid is observed because the light is reflected by t’he mercury. This avoids the complications which are caused by the passage of light through front and rear walls when transmission microscopy is used. I n their article on the microscopy of composite materials, Gracias and O’Dell (68) describe their method for preparing specimens of abrasive papers. The article contains micrographs of cross sections and other views of these composite materials. Riesenberger (205) hris developed t h e met’hod for the electron microscopical identification of metals by electrolytic deposition of t h e metals onto t h e specimen grid. In t’he article reviewed here (205), he describes how he was able to demonstrate this method of specimen preparation using microprojection. The electrolysis of Cu and its conversion to copper picrolonate were demonstrated. TECHNIQUES FOR SPECIMEN EXAMINATION
Photomicrography. Articles relating t o photomicrography can be found elsewhere i n this review. Under I n struments a number of exposure meters are reported (25, 168, 169, 209). Cinemicrographic equipment (26, 160) is also reported under Instruments. An article on infrared color photomicrography (27) is recorded under Infrared and Ultraviolet Microscopy. Szab6 (189) has written a book on
color photomicrography which contains much information of practical value to the microscopist. H e discusses t h e equipment, the procedure for taking color photomicrographs, and t h e procedure for developing color film and color prints. Cowen (3.4)has examined the various fully automatic cameras available for use with t h e microscope and gives a factual summary of t,heir features. T h e spectral sensitivity of “small picture films” used for photoniicrography j s discussed by Hahermalz (71). I n the same article, he discusses light filters and their use for both color and black and white film. Ehrlich (44) has written a careful and complete article about t h e factors which affect t’he fidelity of reversal color film reproduction in photomicrography. Young (210) describes methods for color photomicrography of motile organisms using electronic flash. h fluorescent vital stain is used to reveal the structure of t h e organisms. Freere has developed a cinemicrographic technique for three-dimensional reconstruction from serial sect,ions. Successive sections are photographed on cine film and when shown in a projector, t8he film reveals the three-dimensional structure of the object. X difficulty has been encountered in accurately aligning the sections. I n this article, Freere (55) gives three methods for overcoming t’his alignment problem. Refractometry. Two methods for measuring t h e refractive index of subnanogram particles are described b y LlcCrone and Teetsov (108). When using the Duc de Chaulnes method for determining refractive index, an error is int’roduced by the presence of nonparaxial rays in the imaging system. Miller (120) presents a method of calculating a correction fact’or. With this correction, a n accuracy of 10.002 is attained. Chromy (23) describes a photoelectric apparatus for determining refractive index. H e uses a phase microscope to determine when the refractive indices of the object and the immersion liquid match. The temperature is varied to control the refract,ive index of the immersion medium. Particle Size. Articles describing particle size studies involving holography (63,200, 213) have been reported under the heading Wavefront Reconstruction Microscopy. T h e double-image technique for particle size analysis, its capabilit,ies, and limitations have been discussed b y Barnett ( I S ) . H e also explores new developments in this field. I n a n article by Kaye (921, the efficiency of current stat’istical methods of particle size analysis is explored. Standard definit’ions of “random chord” diameters are proposed. The use of Martin’s and Feret’s diameter is explored, and tech-
niques for improving t h e efficiency of statistical methods are suggested. Tuma (197) has compared particle size d a t a obtained by various dynamic methods with microscopical data. Forty-five samples of fly ashes are compared. The theory is discussed. Stereology, Stereology encompasses t h e methods b y which quantities measured on a two-dimensional section plane are related t o t h e three-dimensional shape of t h e structures in t h e specimen. I n recent years, this field of endeavor h a s become quite popular. Two books dealing with it h a v e been published (36,.45). Both of these books are proceedings of symposia, and contain articles too numerous to be summarized in this review. Humphries has written a chapter in Volume 3 of “Advances in Optical and Electron hlicroscopy” (12) entitled “Mensuration Xethods in Optical A!& croscopy.” This chapter is an extensive review and discussion with a large bibliography. Underwood (198) summarizes stereology, and suggests a systematic nomenclature for the field. H e also gives the basic equations used. Giger and Erkan (65) give the basic stereologic formulas and t,he corresponding sampling t’echniques for the geomet,ric fabric analysis of polished sections of minerals and ceramics. I n another article (66) the thickness error in quantitative transmission microscopy of ceramics is examined. Pe6ar (137) presents a series of formulas relating characteristic dimensions and t’he relative surface of particles. In two articles (151, 154),the basic theory of point’ counting for areal analysis is discussed, and suggesttioils are made for improving the method. I n another related article, iller2 (118) discusses volumetric analysis of sectioned material using lineal and point count methods. H e proposes the use of circular and test figures to overcome the difficulties which arise from preferred orientation in the specimen. Automatic I m a g e Analysis. .A general article eiitit,led “ T h e Niche for Automated Quantitative hlicroscopy” by Ratz (155) has appeared in J f e t a l Progress. A t the present time, automated quantitative analysis usually refers to particle size analysis, b u t rapid progress is being made in more subtle forms of image analysis. One of the chapters in Volume 2 of “Advances in Optical and Electron Microscopy” (11) deals wit,h “Digital Transformation and Computer Analysis of Microscopic Images.” This is a progress report on instruments and techniques which are being developed. An instrument, which is available commercially is the Quantimet Image Analyzing Computer. This instrument was the subject of a series of articles appearing in Microscope. Three articles
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discuss the instrument (48),its proving trials (S), and presentat’ion and classificat’ion of formulas used when applying the Quantimet to the solut’ion of stereological problems (49). The remainder of the articles deal with applications of the Quantimet. They include quant’itative metallographic phase analysis (78), assessment of nonmet’allic inclusions in steel ($a), t’he assessment’ of the growth and separation of deoxidation products in steel ( l o o ) ,the quantitative determination of combined oxygen in oxide inclusions (162),the areal analysis of composite materials (Si), and the quantitative evaluatioii of pigment dispersions (179). Another article describes a technique for staining certain types of transparent particles to facilitate size analysis ( 5 ) . Miscellaneous hliscellaneous inst’rumeiitalmethods of examining specimens include a combined t’ransmission reflection type of illumination for chromosome autoradiography (8),t’he use of the light microscope as an optical diffractometer (62)) and a procedure for using a camera-lucida for low contrast material (50). Huber (81) suggests a method for areal analysis using a photographic enlarger in reverse. The bulb of t h e enlarger is replaced by a photoelectric cell and a print of the specimen illuminated a t a 45’ angle is placed under the enlarger. The reading of the photocell current indicates relative areas of light and dark constituents. Rienitz (159) discusses the negative-positive method of showing relief in microscopical images and compares this technique with schlieren and interference microscopy. T h e technique appears t’o be too difficult to be practical. hlcCrone (104) discusses the applications of hot stage microscopy. -4aronson and Ansell (1) have edited and had published a collection of papers from a n AINE symposium on high-temperature high-resolution metallography. Jones (87) discusses fusion techniques in chemical microscopy using a hot wire technique to produce stable temperature gradients. Vaughan (201) has constructed a microphotometer to record single crystal melting values and other phenomena. This apparatus uses polarized light and is, therefore, suitable for anisotropic mat’erials only. Galjaard (61) discusses the sources of error in the determinat’ion of dry-mass concentration in tissue sections. These include section thickness, immersion media effects, the type of microscope used, and the method of measuring optical path differences. A study of t’he reversible agglomeration of tungstic acid sol particles has been made by Furusawo and Hachisu ( 5 9 ) . They have devised a special cell for t’his type of study. A method of examining mesophase-forming materials by optical microscopy and de-
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polarized light intensity is described b y Rarrall and Sweeney (15). Forlini (51) has expanded and revised the tables for the determination of unknowiis by dispersion staining. APPLICATIONS OF CHEMICAL M I C R O S C O P Y
Crystallography. A general method for investigating nucleatmionand growth processes by quantitative metallography has been reported by Woodhead (207). McFee (109) describes the apparatus and techniques used for the holographic observation of cryst’al growth from the melt. I n a n article on t’he precipitation of solvent-containing pseudo-polymorphic crystal forms and polymorphic modificat’ions of steroid hormones, KuhnertBrandstatter and Grimm (95) discuss thermomicroscopical and infrared spectroscopical studies of 84 steroid hormones. Stay and Reumuth (184) have studied the effect of the presence of a non-ionic surfact’anton crystal habit’. Rosen (164) reports on the polymorphism of l,l,l-t~rinitroet~hane and hexanitroethane. Both materials exhibit eiiantiotropic polymorphic t,raii:f ormations. Kantz and Desando (90) report’ the physical, optical, and X-ray crystallographic properties of diphenylsilanediol. Uarrall et al. ( 1 4 ) describe the polymorphism of cholesteryl myristat’e. They used differential thermal measurements as well as microscopy in the study. Eiinulat (47) discusses the phase identification of mesomorphic states and present’s a scheme of analysis. Resins and Polymers. T h e light microscope was used by Pearson (136) to study the st’abilization of poly(viiiy1chloride) by lead salts. Xicroscopical techniques are combined with infrared absorption spectroscopy, emission spectroscopy, and X-ray diffraction to identify trace contaminants in polymers (63). F r y et al. (67) have applied polarizing microscopy t,o the st.udy of the microstructure of glass-reinforced plastics. Textiles and Fibers. A scheme for identification of synthetic fibers b y measuring their specific birefringence has been proposed b y Netteliistroth ( 1 2 7 ) . He has applied this scheme to the identification of poly(amide), poly(ester) and poly(acrylonitri1e). Poling and Hollies (146) have discussed special applications of interference transmission microscopy to fiber problems. Specifically, they studied the impregnation and coating of fibers with polymers, the measurement of the thickness of microtome sections of fibers and small variations of refractive index within fibers. The refractive indices and birefringence of fibers can be measured accurately using channelled spectra from the fibers in a n interference microscope
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(110).
ANALYTICAL CHEMISTRY, VOL. 42, NO. 5, APRIL 1970
Wood and Paper. Page (132) has devised a method for determining t h e fibrillar angle in wood tracheids by filling t h e lumen of t h e fibers wit’h H g and examining t h e m using reflected polarized light. Quackenbush (148)discusses the analysis and solution of two problems encountered in a paper mill. These problems were concerned with the causes of flaws in poly(ethy1ene) coated papers. Minerals and Ceramics. Xpplications of microscopy to t h e s t u d y of minerals and ceramics have been reported elsewhere in this review under Staining and Et,ching (18, 133), under St,ereology (65, 66) and under Techniques for Specimen Preparation, Miscellaneous ( 2 ) . Books concerned with the microscopy of minerals and cera,mics include: “Ceramic Microstructures: Their Analysis, Significance and Production’’ ( 5 8 ) , “Die Rohstoffe der Keramik” (150) and “Phase Diagrams for Ceramists” (98). The second of these books deals with silicate structures, the others are more general. The last book mentioned is a supplement to the basic volume of the same title which was published in 1964. Techniques for t,he preparation of polished thin sections using three stages of polishing are described by Moreland (123). Trojer (196) discusses the application of microscopy in cement and plaster technology. His article contains numerous micrographs of thin sections and powder preparations. I n an art’icle entitled “Microscopic Exaniinat’ion of Fused Cast Alumina Refractories,” Coss (33) discusses the identification of cy and p A1203 by transmitted and reflected polarized light microscopy. Gracias and O’Dell (68) report on a study of abrasive papers using microscopical techniques. The etching of barite to reveal dislocations is reported by Pat’el and Koshy (133). Foivkes and Parton (62) have identified sulfur and iodine condensates in bubbles in glass by examining thermal phenomena which occur on heating the glass. Metals. h l l of t h e articles coiicerned with metallography have already been reported under Grinding and Polishing (56, 7 0 ) , under Microtomy (69),under Staining aiid Etching (133, 185) and under Interference Xicroscopy (193j. Forensic Microscopy. A series of articles dealing with forensic science has appeared in A p p l i e d Optics. T h e titles of these articles are as follows: “Forensic Opt’ics” (a&?), “Applied Opt.ics in Crime Detection” ( S I ) , “Optics and Instrumentation in Forensic Science” ( f o g ) , “Optics in Pathology” (85),“Radiation aiid Optical Techniques in S’isual Examination of Paintings” (M),and “Investigations of Historical Objects Ktiliziiig Spectroscopy and Other Optical Methods” (129). ,inother
article, not in this series, entitled “Proof in Criminal Cases by Means of hpficroscopical Evidence” has been published b y hfart’in (114). ANALYTICAL MICROSCOPY
bide phases of (U, Pu)C and PuSi precipitates in (U,Pu)C fuels can be identified. Wiesenberger (206) describes a microprojection apparatus which he used to demonstrate the electrolytic deposition of C u and the conversion of copper to the picrolonate. These are reactions which he uses for chemical electron microscopy. Kuhnert-Brandstatter and RIuller (96) report on their experiences in the hot stage microscopy of inorganic compounds. They used t h e Leite 1350 “C hot stage t o determine melting points, eutectic melting points, and binary phase relations.
RIany articles describing applications of microscopy to chemical analyses have been reported under various headings earlier in this review. Under the heading Refractometry, there are three articles (29, 108, 120),under Techniques for Specimen Preparation, Miscellaneous (188),under Crystallography (14, 47,90, 95), under Text’iles and Fibers (127) and under Minerals and Ceramics (33, 6 2 ) . Nearly all of t h e articles reported under t’heheadings Stereology and h u t o LITERATURE CITED matic Image Analyses are concerned (1) Aaronson, H. I., Ansell, G. S., “Highwith quantitative chemical analysis. Temperature High Resolution hletallography,” Gordon and Beach, ?Jew Part’icleSize also is a specialized type of York, N. Y., 1968. quantitative chemical analysis. (2) Adams, M. D., Tong, S. C., ANAL. The book on “1Iicroscopical IdentifiCHEM.,40,1762 (1968). cat’ion of Organic Compounds” (186) is (3) Allmand, T. R., Microscope, 16, 163 (1968). concerned with microscopical met’hods (4) Alpiar, R., A p p l . Optics, 7,1461 (1968). of analysis by precipitation of charac(5) Amor, A. F., Block, &I., J . R o y . teristic crystals. Reversible complex Microsc. Soc., 88, 601 (1968). formation also may be used as a n aid in (61 Anderson. W. L.. J . Opt. SOC.A m e r . . ‘ 59, 224 (1969). ’ identifying materials. Scullion et al. (7) Arregger, C. E., J . R o y . Aficrosc. SOC., (174) have used complex formation be87, 191 (1967). tween AD;I (acetdimethylamide) and (8) Back, F., Dormer, P., Zeiss-Mitt. HRIX (1,3,5,7-tet~ranitro-lJ3,5,7-tetra- Fortschr. Tech. Opt., 5, 80 (1969). (9) Barer, R., “Lecture Notes on the Use azocyclooctane) for identification of of the Microscope,” 3rd Ed., Blackwell either reagent. H1IX also forms comScientific Publications. Oxford and plexes with a number of other comEdinburgh, 1968. pounds. The identification of materials (10) Barer, R., Kature, 217, 672 (1968). (11) Barer, R., Cosslett, V. E., “Advances in cells is the subject of two articles (64, in Optical and Electron Microscopy, 83). Genest et al. give reagents for the Vol. 2,” Academic Press, New York, identification of reticuline, isoboldine, N. Y., 1968. laurotetanine, laurolitsine, and isocory(12) Barer, R., Cosslett, V. E., “Advances in Optical and Electron Microscopy, dine-N-oxide. Shead (175) reports on Vol. 3,” Academic Press, New York, the conditions for growing well formed N. Y., 1969. crystals of canbhardin b y sublimat,ion. (13) Barnett, M. I., Ann. N . Y . A c a d . Sci., The crystals can then be ident’ified by 158, 674 (1969). (14) Barrall, E. )I., Porter, R. S., Johnson, profile angle measurements. J. F., Mol. Cryst., 3, 103 (1967). The identification of particulat’e mate(15) Barrall, E. &I., Sweeney, M. A , , ibid., rials has become very important in 5, 287 (1969). recent’ years. McCrone and his co(16) Bayard, SI.,Reffner, J. A., Microscope, 17,287 (1969). workers have published a number of (17) Beck, J. L., A p p l . Opt., 8, 1503 articles dealing with setting up a lab1. oratory for particle ident,ification (40, (18)1969 Bender, J. H., Metallography, 1, 19 107) and with the t’echniques and tools (1968). ._ (19) Bennett, A. J. R., J . S c i . I n s t r u m . E , for particle identification (38, 105). Series 2, 2, 819 (1969). “The Particle .Itlas” by IfcCrone, (20) Beyer, H., J. R o y . Microsc. Soc., 87, Draftz, and Delly (105) is the standard 171 (1967). reference for those coiiceriied with the (21) Bishop, A. C., “An Outline of Crystal hlorphology,” Hutchinson, London, identifieation of particles. 1967. H. F. Schaeffer has published three (22j Blank, J. R., Microscope, 16, 189 articles on analytical microscopy (170(1968). 172). A11 of them are concerned with (23) Bradbury, S., J . Sci. Instrum., E , Series 2, 1 , 3 (1968). bhe identification of the platinum metals (24) Bradbury, S., Microscopy, 31, 1 and gold. The reagents are 10-methyl(1968). acridinium chloride (170), 2-amino(25) Brown, hl. S., Mikroskopie, 23, 173 quinoline (171), and lJ5-napthyridine. ( 19681. (26) Burton, A. L., J . Microscopy, 89, 186 Bender (18)describes a scheme for the (1969). qualit’ative analysis of metallographi(27) Casida, L. E., Jr., Science, 159, 199 cally prepared specimens of uranium (1968). plut’oiiiuni mixed carbide fuels. h (28) Champagne, E. B., Massey, S . G., A p p l . Opt., 8, 1879 (1969). three-step technique was used consist,ing 129) Chromv. S.. A m e r . Mincralooist. “ , 54 , of electroetchiiig, microhardness mea549 (19693.‘ ‘ surement, and preferential etching, (30) Cocks,’G. G., AN.AL.CHEM.,40, l58R T h e metallic dicarbide and sesquicar(1968). I-_
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(31) Conrad, I. W., A p p l . Opt., 8 , l (1969). (32) Cook, W. F., J . Roy. Micros. SOC., 88. 607 (19681. (33) ‘COSS, ~ P .J. , ~ u s t Ceram. . SOC.,2, 53 (1966). (34) Cowen, G. C., Proc. R o y . Microsc. SOC.,4,71 (1969). (35) Davidovits, P.,Egger, M. D., Nature, 223, 831 (1969). (3f3) DeHoff, R. T., Rhines, F. N., Quantitative Microscopy,” RlcGrawHill, Kew York, N. Y., 1968. (37) Delly, J. G., Microscope, 17, 193 (1969). (38) Ibid., p 291. (39) Ibid., p 205. (40) Delly, J. G., McCrone, W. C., Contam. Control J., September, (1967). (41) Dodd, J. G., Microscope, 17, 1 11969). (42) Edholm, P., J . R o y . Microsc. SOC., 88, 351 (1968). (43) Ehlers, E. G., J. A m e r . Ceram. SOC., 52, 451 (1969). (44) Ehrlich, S., Microscope, 17,97 (1969). (45) Elias, Hans, “Sterology, Proceedings of the Second Int’l. Congress for Steralogy, Chicago, 1967,” SpringerVerlag, S e w York, Inc., 1967. (46) Engle, J. L., Freed, J. J., Rev. Sci. Instrum., 39, 307 (1968). (47) Ennulat, R., M o l . Cryst., 5, 405 (1968). (48) Fisher, C., Cole, >I., Microscope, 16, 81 (1968). (49) Fisher, C., Nazareth, L. J., ibid., p 95. (501 Forer. A.. J. Rou. Microsc. SOC.. , 88., ‘ 611 (1968). ’ (51) Forlini, L., Microscope, 17, 29 (1969). (52) Fowkes, A. J., Parton, C., Glass Technol., 10, 147 (1969). (53) Fourney, ill. E., RIatkin, J. H., Waggoner, A. P., Rev. Sci. Instrum., 40, 205 (1969). (54) FranFon, M., “Einfuhring in die neueren Methoden de Lichtmikroskopie,” Verlag G. Braun, Karsruhe, Germany, 1967. 1553 Freere, R. H., Microscope, . . 16,. 235 (1968). ’ (56) Freund, H., “Die metallographische Proben-Praparation fur die mikroscopische Untersuchen,” Umschau Verlag, Frankfurt, Germany, 1968. (57) Fry, J., Butler, B. C. M., Holland, R . A. G., Proc. R o y . Microsc. SOC.,3, 60 (1968). (58) ,Fulrath, R. &I.,Pask, J. A,, “Ceramic blicrostruct,ures : Their Analysis, Significance and Production,” John Wiley & Son, Inc., New York, N. Y., 1968. (59) Furusawo, K., Hachisu, S., Scz. Light ( T o k o y o ) , 16,71 (1967). (60) Gabor, D., Optik, 28, 437 (1968). (61) Galjaard, H., J. R o y . dficrosc. SOC., 87, 157 (1967). (62) Gall, J. G., J. Cell. Sci., 2 (1967). (63) Garrett, H. L., Traylor, P. A,, Microscope, 16, 295 (1968). (64) Genest, K., Lowry, L. J., Hughes, D. W., Aficrochem. J., 14, 249 (1969). (65) Giger, H., Erkan, Y., Z. Wiss. Mikrosk., 69, 36 (1969). (66) Gokularat,hnam, C. V., Freiman, S. W., DeHoff, R. I., Hench, L. L., J . A m e r . Ceram. Soc., 52, 327 (1969). (67) Goldstein, D. J., J. Microscopy, 89, 19 (1969). (68) Gracias, C. E., O’Dell, J. A., i?fzcroscope, 17, 161 (1969). (69) Graham, D., Rev. S c i . Instrum., 40, 897 (1969). (70) Gray, R . J., Metal Progr., 133 (1968). (71) Habermalz, F., Mikroskopie, 22, 62 (1967). (72) Hamer, D. H., Microscope, 17, 137 (1969). (73) Harmer, P. R., Ford, S. D., Dugdale, R . A,, J. Sci. Instrum., E, Ser. 2, 1, 59 (1968). ~
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(101) Loasby, R. G., J . Sci. Instrum., Ser. 2, 2, 148 (1968). (102) Lucas, D. 3I., A p p l . Opt., 8, 15 (1969). (103) McCrone, W.C., Laboratory M a n agement, January 1968. (104) McCrone, W. C., A m e r . Lab., Xovember 1969. (10.5) McCrone, W. C., I h f t z , R. G., Delly, J. G., “The Particle Atlas,” Ann Arbor Sci. Publishers, Ann Arbor, hIich., 1967. (106) RIcCrone, W. C., Draftz, R. G., Delly, J . G., “The Particle Analyst,” Ann Arbor Sei. Publishers, Ann Arbor, hlich., 1968. (107) McCrone, W. C., Hertrich, J. A , , Microscope, 17, 77 (1969). (108) McCrone, W. C., Teetsov, A., ibid., p 83. (109) bIcFee, R. H., J . A p p l . Phys., 40, 3873 (1969). (110) lIcKee, A., Woods, H. J., J . R o y . Mzcrosc. SOC.,87, 185 (1967j. (111) Magill, H., Gunning, B., J . Nicroscopy, 89, 217 (1969). (112) Magill, P. J., Wilson, A. D., J . A p p l . Phys., 39, 4717 (1968). 122 R
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Nucleonics w. S. lyon, E. Ricci,
I
and H . H. ROSS, Analytical Chemistry Division, O a k Ridge National laboratory, Oak Ridge, Tenn. 37830
first review four years ago \$-e stated our intention to restrict our discussion to “items t h a t appear original, novel, or potentially useful.” R e continue to use this guideline, b u t in a period marked by considerable disenchantment with hard science, particularly the physical and chemical, and most particularly those aspects of it which seen1 to bear little relation to current pressing social problems, it is not surprising t h a t we find a smaller number of papers that meet’ our criteria. I t appears that nucleonics has reached maturity and will probably yield fewer startling innovations than in the past. Cert’ainly many applications and routine methods continue to be published, and it’ is, we think, a n iiidicat’ion of the vitality of the field t’hat these occur. B u t the innovative progress of most tracer applications, measurement techniques, and radioanalytical methods has been slower and less spectacular than one might wish. This is true for neutron activation also; both generator and reactor applications have increased, but that which is original and of non-routine interest has appeared primarily in the fields of charged particle and nuclear reaction analysis. T h e genuine interest of nuclear scientists in attempting to apply their methods to noli-nuclear problems is a healthy sign, and though this somewhat limits our review, we applaud the effort to make nucleonics relevant to today’s problems. If this trend toward application continues, the reader might appreciate a more application oriented review. Sucleonics continues to be a strong, vigorous, and exciting field in which to work. T h e technological accomplishment of the decade-landing men on the moon-was followed by return of lunar samples to the Lunar Receiving Laboratory which was designed and directed by scientists and engineers trained in the nuclear field. The first N WRITIXG OUR
measurements on the lunar samples were made by nuclear scientists a t the space lab (126), and samples from both Apollo 11 and 12 have gone to radiochemists and activation analysts all over the western world. Turning toward earth, we see t h e challenge to nuclear scientists as one of making our discipline relevant and useful in the solution of biological and environmental problems a t home. As noted below, some success in these efforts has already occurred. iit least three pertinent new journals have appeared: A c t i n i d e s R e v i e w , R a d i o chemical a n d Radioanalytical Letters, and J o u r n a l of Radioanalytical C h e m i s t r y ; the latter has a special bibliography section. A11 the journals are bubbling over with papers, and from them we have attempted to choose those t h a t seem most closely to meet our criteria. Table I, as usual, lists books, reviews, and printed matter of general interest. Some material that we might normally have covered in this article has been the subject of special reviews elsewhere. Rather than duplicate or cursorilj cover such work we have listed the reviews in Table I and omitted detailed discussion of these subjects from our text. h good general source for a wide diversity of isotope applications is the quarterly publication Isotopes a n d R a d i a t i o n Technology where extensive reviews and bibliographies can be found. Absolute Measurement of Radioactivity. Some of t h e problems asso-
ciated with choice of a method for absolute measurement are discussed in a paper concerned with current efforts in activity determinations (8). I n a method of absolute standardization using liquid scintillation counters, the observed output was compared with that produced theoretically from the beta ray spectrum for a number of nuclides; activities determined for three low energy emitters: tritium, carbon14, and chlorine-36 agreed well with standards (84). General information
concerning standards, accuracy, certification, and types available are described in a publication from the Radiochemical Centre, hmersham (142). A table listing 93 radionuclides for which standard preparations are available was published; also included is information concerning sources of supply and use (166). The same author is now engaged in a Y R C sponsored project surveying the needs and interest in radioactivity standards, as well as
Table I. Books, Reviews, and Bibliographies
A . General-23, 53, 58, 64, 119, 136, 166, 177, 184, 207 B. RadioisotoDes in Chemical Analvsis Radio-reiease-40 Chromatography-29, 194 Isotope dilution-26, 87 Elect roanalysis-3 Isotope effects-215 Separations-94 Mossbauer-89 Fluorine77 Environment-46, 60, 96, 202 Geology-70, 107, 128, 161 Biology and Medicine-9.3, 220, 12.3, 146, 203 Polymers-6, 39 AerosDace-lbS Rletailurgy-17 Oil-154 Natural gas-75 Solids-1 2 C. Radiochemistry General-176. 216 St.andards-43, 124 Transuranium-7, 13, 31, .96,61, 134, 138
Elements-Pa 90; Tc 109; C 163; Si 182 Labeled compounds-16, 42, 193 D. Industrial-19, 199, 205, 206 E . Instrumentation-18, 104, 160, 188, 200 F . Activation Analysis General-129. 163 Charged Particle and Photon hIethods-9, 36, 132 Forensic Applications-78, 98, 131 Aids--Nuclear Reaction Tables-I 15 Photon Activation Products-
io, 11
Neutron Reaction Gamma Rays-15, 71
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