Practical flor cytometry

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R.A et al (1993)

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bochem

Porosrtoi. 62, 2333242 Dluzewski. A.R. et 01 (I 992) j Cell SCI 102, 527-532 Ward, G. et oi (I 993) 1 Ceil Scrence 106. 237-248 Nash, G.B. er (II. (I 989) Blood 74, 855586 I Cranston, H.A. et al. (1984) Scrence 223, 40&403 Perlmann, H. et al ( 1987) /. Clan.Mlcrobroi 25, 234772354 Aikawa, M. and Miller, LH (1983) in M&no ond the Red Ceil (Evered, D. and Whelan. J eds). pp 45-63, Pitman Foley, M. et 01. ( 1990) Mol. Blochem. Porasrtol. 38, 69-76 Foley, M. et al (I 99 I) Mol Blochem Poros~toi 46, I 37- I48 Ruangiirachupom. W et ol ( I99 I ) Exp Parosltol 73, 62-72 Favaloro. JM. et al. (I 986) Nuclerc Acrds Res !4,8265-8277 Foley, M. et OI (1994) Exp Porosltcr 79, 34&350 Bark P. et al ( 1992) Trends Blochem SCI I7 129 Langer, T. et 01 ( 1992) Nature 356, 683-689 Da %a. E. et a/. (1994) Mof Brochem Pcrasrtol. 66, 59-69

Practical Flow Cytometry Howard M. Shapiro,john Wiley & Sons, I995. f49.95@xxviii + 542 pages) ISBN 0 471 303 763 The development of quantitative analytical methods applicable to lndivtdual cells or cell organelles is important for the progress of the understandlng of cell biology The accurate quantlficatlon of different cell types has beer advanced in recent years by the use of flow cytometry. Sensltlve, specltic and rapid measurements can be made, using a flow cytometer, when cells are labelled with fluorescent molecules In this respect, flow cytometry has inherent advantages over other techniques such as microscopy. In a flow cytometric system, large numbers of cells or particles flow within a fluid stream In a single file past a laser beam, where they are lndlvidually evaluated. If the cells have been bound by a reporter fluorescent molecule, the fluorochrome will transiently absorb the laser light and then emlt it at another light frequency. The flow cytometer IS equipped with detectors for this emitted light, which not only detect the fluorescent signal but quantlate its intensity. The intensity IS directly proportional to the amount of the fluorescent compound bound to the cell or particle. Evaluating large numbers of cells quantitatively and reproduclblv bv

22 Sherman,

I.W

et a

161-178 23 Vlal, H.J. and Ancelln. 24 25 26 27 28 29 30 3I 32 33 34

(1992) ML

t31ol. Cell 74

( 1992)

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Brochem 18. 259-306 Tllley L. et a’ (1990) Bochrm i31ophys Acto 1025, I 355 I 42 Crandall I. et al. (1993) Proc Not1 Acad So. USA 86,9089-9093 Land, K.M. et al. ( 1995) Pcrosrtology Today I I, 19923 Behan R. and Haldar K ( 1994) Exp Porosrtol. 79, 25&259 Hobbs. AR. and Saul. A.J ( 1994) EIxp Parasttol 79, 260-269 Coppel, R.L et oi cI9843 Nature 3 IO, 7899792 Coppel, R.L et 01 (I 986) MO/ Blochem Pora srtol. 20, 2655277 Lustigman, S. et 01 ( 1990; MOI Blocherr Porasltol. 38, 26 l-270 KlleJlan, A et 01 (I99 :I MY Blochem Porasitol 44, I75 - I82 Chlshtl. AH et al ( 19921 MO/ Blochem Parasfto1.52, 283.-288 KlleJlan, A. ( 1979) Proc. Nati Acod Scr USA 76,

4650-4653 35 Leech JH. et of (19841 j txp 1567-1575 36 Howard, RJ. et ai. ( 1990) Am j

Med

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flow cytometry Increases the statistical confidence and precision of data. Pracrlcai Flow Cytometry IS a very useful book on this complex but vet-y widely used technology. In the preface to the first editlon Shapiro says: ‘This book can tell you almost everything you need to know about flow cytometry what t IS, how It works, what you can do v&h It and how to buy or build and use flow cytometers’ Indeed, the book describes in detail the basics of flow cytometry with chapters on ‘How flow cytometers work ‘Data analysis’, ‘Flow sot-tlng’, ‘Buying flow cytometers’ and ‘Using flow cytometers’. An extended (very InformatIve, clear and coherent) chapter deals wtth a large list of parameters and probes. This chapter In par-tcular gives a good Insight Into what you can do with a flow cytometer and the physlcal and/or chemical charactenstlcs of cells which can be measured The most popular applications are In the fields of lmmunology and DNA content/cell cycle analysis The immunological appllcatlons, such as leukocyte dlfferentlal counting and lmmunophenotyping are now quite routine. From the chapter on ‘Parameters and probes’ it IS clear, however, that flow cytometry as a field continues to expand rapldly. Extensive progress In the development of new probes and methods, as well as new appllcatlons, has ocrilrred durmg the

Hyg. 43, 15-29 C.F. et al. (1991) Proc. Nat1 37 Ockenhouse, Acod. Ser. USA 88, 3 175-3 I79 EMBO 1. 12. 38 Hen-era, S. et al. (1993) 1607-1614 39 Pasloske. B.L. et a/ (I 993) MO/. Brochem. Porasitol. 59, 59-72 40 Helmbly. H. et oI (I 993) Infect. Immun. 61, 284-288 41 Shen, 8. et 01.(I 985) 1, Ceil Brol. 102, 997- I006 42 Gruenberg, J. and Sherman, I.W. (I 983) Proc. Noti Acad Scr USA 80. IO87- IO9 I 43 Borst. P. et 01.Ceil (in press)

Note added in proof A senes of papers has recently been published that identify PRMP- I as a large gene family of variable sequences Implicated In cytoadherence and antlgenic vanatlon43. Michael

Foley and Leonn TJley ore at the

School of

Brochemrstry,

La

i-robe

Unrvenrty,

3083, Australia. Tel: +61 9479 2158, Fax: +61 9479 2467, e-mail:

Bundoora,

Victoria,

[email protected]

past few years. Numerous new methods have been Introduced and applied in a variety of fields. The long list of parameters, presented by Shapiro, which can be measured by flow cytometry using fluorescent probes Includes: DNA and RNA content, chromatin structure, antigens, lipids, surface sugars, membrane organization and permeability, endocytosis, oxidatlve metabolism, DNA degradation (as In apoptosis), enzyme activity, membrane potential. These parameters give a good impression of the wide vanety of possible applications of flow cytometry in the study of cell biology. Apart from the wealth of basic Information, I appreciated the critical assessment of the applications in different fields of research. However, the chapters describing the specific applications of flow cytometry contain less detail on practical aspects, although many useful references are given. Therefore, the book may be of less immediate value for researchers who want to use flow cytometry for a specific purpose and look for experimental detail of already established methods. Other books on flow cytometry might then be more helpful, for example Methods in Cell Biology: Flow Cytometry’, where many different applications are described in detail in a manual-like format. The short chapter on applications within the field of parasitology lacks expenmental detail, but It does give a 439

Books comprehensive oven/lew of papers or flow cytometry rn parasitology. For example, the papers on the use of flow cytometry in malana research (detection of parasites, analysis of drugs, red blood cell antigens) and In iryponosomo and iedtmonra research (DNA content, growth and surface characterIstics, effects of drugs etc.). Shapiro ends this chapter as follows: ‘There were a bunch of other papers on flow cytometry In parasitology which I have Included in the references; since I’d IIke to move on, I won’t open that can of worms and protozoa. I wtll. Instead. suggest that you run a copy of thus sectIon by the pat-asitologlsts in your institution. especially if they don’t use your machlne. Take them to lunch. Buy

Parasites in Human Tissues by Thomas C. Orihel and Lawrence R Ash, American Society of Clinical Pathologists, Chicago, I 995. $ I6500 (xxi + 386 pages) /SBN 0 89 I89 3 72 2 Medical students at Oxford Unverslty have long been taught that anythlng strange in a pathologlcal specimen IS either something that a doctor has put there or a parasite. As to whether or not It IS a parasite and, If so, what kind it IS, there need no longer be any doubt. The aim of tnls lavishly produced book IS to provide pathologists and others with a comprehenslve guide to the identification of parasites found in human tissues. The task facing pathologists IS enormous as the number of different kinds of parasite that might be encountered IS a staggenng 20 protozoa, 63 helmlnths and ten arthropods (and these figures do not include those that cannot easily be ldentlfied at the species level) All the common parasites, and virtually all the uncommon ones, are illustrated here; indeed, there IS so much that even parasltologlsts might be puzzled by the names Bolamuthia, Mammomonogomus and Rictoiaria. They WII be d sappointed, however. not to f;nc sly mention of the tncreaslngly common Bobesro species known to nfect humans, even though these are not, strictly, tissue parasites There is a lot that parastologlsts can take for granted and they snould be able to use the time-tested technique of hopefully flicking through the pages until something familiar catches the eye. On the other hand, the non-para440

them acceptable beverages You might gain fnends and or paying customen’.

From this comment you can already taste that this book IS more suited for flow cytometer operators or researchers running a department In which flow cytometry IS applied to a wider variety of research areas than for parasitologists. (Several parasitologists from our laboratory are already such paying customers who regularly visit the Flow Cytometry Department in the Unlvenlty, using the flow cytometer, for example, for lmmunophenotyplng of host cells in filarla research and for DNA synthesis measurements in malana drug-susceptibility research. Alas. we’re still waiting for that bever-

age.)

sitologlcal user IS guided through the labyrinth of posslbllitles by means of (I) a quick reference table, llstlng each parasite against each organ, and (2) well-referenced Introductory secttons, covering IIfe cycles, geographlcal distribution and cllnlcal features, prefacing each group of illustrations. There are nearly IO0 clInical lllustratlons and over 640 rrlcroscopic sections, all large enough to be seen easily and nearly all of a superb quality. Several different stalntng methods are Illustrated and even the vagaries of haematoxylln and eosln staining do not detract from the excellence of the dtagnostlc features Illustrated, which are clearly described In the accompanying text There are also lllustratlons of a number of artefacts that might be mistaken for parasites at first glance This book IS remarkably up to date, In particular the references, and, away from the pathology department, could serve to supplement more general parasitology textbooks, perhaps encouraging Instructors to use stained sections containing parasites as part of their practical courses. The ar-tefacts might also tempt unscrupulous instructors to test more than students’ knowledge. No matter how beautiful a book is, the real question IS. ‘does It work?’ This one does, In my hands at least, although one has to be careful not to reach any conclusions too quickly, and I nearly fell Into a number of traps. Several different nematodes look superficlally similar and some protozoa, such as the mlcrosporldlans and cyst-forming coccidla, present a challenge to all but the most experienced observer. Apart

For parasitologists with a broad Interest in the general cell biology of their study objects, and for those pat-asitologlsts who want to modify already existing flow cytometric techniques, or to extend their applicability to other cell systems, much useful and interesting information can be found in Shapiro’s book. Reference

I Darzynktewlu, Z., RobInson.JP and Crissman, H.A. (I 994) Methods in Cell Biology,Vol.42, Flow Cytometry (2nd edn), Academrc Press Chris Janse

Laboratory for Parasitology University of Leiden Wassenaarseweg 62 PO Box 9605 2300 RC Leaden The Nethenands

from the very obvious cases, it would probably be best for anyone who is at all uncertain about what they are looking at to seek more expert advice. Another point IS that the specimens Illustrated are the best that are available and a busy histology technician is unlikely to produce material as good as this unless given guidance as to what is required. Most of the photographs seem to be of wax sections, and obtaining the same quality with ctyostat sections would be an achtevement in itself Atthough primarily focused on human ttssues, the parasites illustrated, or similar forms, are frequently found in domertlcated or experimental animals. Postgraduate students would benefit from a familiarity with the contents of this book, which certainly deserves to be seen and used outside pathology laboratories. Finally, I have no doubt that fabnc deslgnen and artists could also find inspiration here, which is something that cannot be said of any other parasltological book! Frank Cox Drvisron of Life Sciences King’s College London

Campden Iill Road London, UK W8 7AH

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