BRAINS AND BRAIN PRESERV A.TIVES. By Ales Hrdlicka,Assistant Curator, Division of Physical Anthropology.PART I.PHYSICAL CHANGES IN HUMAN AND OTHER BRAINS COLLECTEDUNDER DIFFERENT CONDITIONS AND PRESERVED IN VARIOUSFORMALIN PREPARATIONS.Anatomical and anthropological investigations on the brain todetermine the homologies and differences in the organ and all its parts,between man and other animals, and between races and other groupsof mankind, make large collections of brains necessary. Such collec-tions imply the use of means b}^ which the brains can be kept indeli-nitel}' in good condition for study. An ideal means would be. onewhich would allow" every specimen to preserve its form, size, weight,and all macroscopical as well as minute features. The need for suchan agent has long been felt and led from dry preparations to the useof various liquid preserv^atives, among which, subsequently to theintroduction of that chemical in 1S9I: by Blum, have been solutionscontaining formaldehyde.The commercial solutions of formaldehyde, known ordinarily asformol, or formalin, have, even when much diluted, the qualitv" ofrapidly penetrating and hardening brain tissue, allowing but littlealteration in the form of the organ and preserving much of its color.Furthermore, when hardened, specimens can be kept in the formalinsolution without further noticeable change quite indefinitely and thepreservative is not expensive. The chemical, however, is not wholl}"without objections; some persons are affected adversely by its fumes,the volume and weight of the brain are increased somewhat in itssolutions, and it does not serve best the purposes of histology; yet theother advantages of formalin are so great that, until something moreefficient be discovered, it can not well be dispensed with for brainpreservation. . ? ,Proceedings U. S. National Museum, Vol. XXX?No 1451. 245 246 PROCEEDINdS OF THE NATIONAL MUSEUM.Effoj-ts have been made to correct the faults of formalin b}- theaddition of other substances to its solutions, or l)y following- these,after the desired hardening of the brain had been eti'ected, with otherpreservatives. Tt has been combined with or followed by various pro-portions of alcohol (Parker & Floyd, Marie, Gerota, etc.), potassiumbichromate, or Miiller's fluid (Diedrichs, G. Retzius), glycerin (Lan-zillotti-Buonsanti, Chencinski), sodium acetate with sodium chlorideand alcohol (Stroud, Wilder), sodium chloride and zinc chloride (Fisk),sodium chloride alone (Spitzka). and bichloride of mercury." All ofthese combinations have been reported upon favorably. The eti'ectsof several formalin solutions have been observed * with some detail,but of no single solution do we possess exact and sufficiently detaileddata as to its action on the ])rain, especially' physically, and its actionon the brains of persons of different ages, or on those of diflerentanimals, or rinall}' on those collected under widelv different conditionsof the organ, or of temperature. Yet it is important to be acquaintedwith such facts. It is desirable to know which really is the best solu-tion or combination for at least most of the specimens, so that such apreparation alone may be used. Such knowledge would tend to bring-about not only a much-desired unit}^ of procedure, but also a generalundci-standing, at any stage, of the state of our material, so far asformalin preservation is concerned. The use of a single solution withwell-known effects would regulate our records and methods, and allowof a degree of accuracy in weight determinations and measurementsnot now possible.With these facts in mind, and remembering the excellent work l)vDonaldson in 1894 on the physical changes in the brain produced byvarious preservatives in use before the introduction of formalin, thewriter, in establishing a brain collection in the Department of Anthro-pology of the United States National Museum, has endeavored to makea series of tests with several solutions, the main component of whichwas Merck's formalin.The material accruing to the ])rain collection of the Museum ishetei-ogeneous, ranging from man's braius'' to those of the lowestmanunals,'' and from aged individuals to embr3"os, hence it was par-ticularly suitable for experiments. Besides this it is alwaj^s possible ? Still other {-orapositionH were employed for the purposesi of histology and pathol-ogy?see Huiiimary in TellyeHnitzki. Special methods, also, having no bearing onthe theme of this paper, were devised for the j)reservation of the natural color ofvaiioiis organs. ^ See Dexler, p. 382, after Flataii; records of the weight of the brain in 1 per cent,5 per cent, and 10 per cent formalin solutions in 1, 3, 30, 1)0, 150, and 450 days.?Of these, unfortunately, not a sufficient number were received in good conditionduring the progress of the experiments. ''The term "mammal" is used, for want of a better term, throughout this i)ai)eras a designation f(jr other mammals than man. NO. 1451. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 247in a city to obtain in fresh condition large numbers of heads ofslaughtered animals. Utilizing both resources, a double plan was fol-lowed. A number of ditferent formalin solutions was made up, somein concentrations used l)y other workers and a few empirically as tostrength, and each solution was used on a series of brains as the}^ werereceived, including specimens of every nature. The second proced-ure was to obtain a large number of brains, as far as possible in thesame condition, from some one fair-sized animal, and to subject uni-form series of such brains to the action of different solutions. Theresults of this latter inquiry appear in the second part of this paper.There are numerous factors which, as Donaldson has already shown,aft'ect the changes in the brain in the same solution. One of these isthe degree of freshness of the brain; another is the temperature ofthe air (large differences); and still another is the presence or absenceof the soft membranes. Only the last of these conditions was capableof being fully regulated in the National Museum collection. Thesubjects from which brains are here obtained come from differentsources, and it is impossible to get all the brains equally fresh; and asto cold and heat, the collecting continues throughout the year, and thelaboratories are not so fitted as to keep up an even temperature. Yetno specimens were included in the tests that were sufficiently advancedin decomposition to make their hardening and preservation doubtful;and the changes of temperature in the laboratory where the brain col-lection is stored would not exceed 40? F. as the maximum in thecourse of the year. The brain w^as always laid into the preservativewith the soft membranes intact or but slightly injured.The regular procedure in cases of the first category was as follows:The brain, being extracted without the dura mater, was immediatel}^Aveighed; the solution in which it was to be laid was prepared before-hand; a layer of absorbent cotton was placed on the bottom of theglass jar to be used, and a quantity of the preservative poured in; thebrain was then placed into the solution, with its base dow^nward on thecotton, so as to rest easily (the cerebellum and cerebrum in the largerbrains being separated by a thin layer of cotton), and a sufficient quan-tity of the preservative was added to rise 1 to li inches above the speci-men. The jar was then closed, labeled, and placed on a shelf, whereit remained for one week. No injection through the arteries or intothe ventricles was practiced, because it would have been impossiblewith all the specimens, and it was not found essential. On the eighthday the brain was taken out, drained in a fixed manner, and thenweighed; the old cotton and solution were replaced with new, in samequantity, the brain was put back into the jar and placed again on theshelf. One month after receiving the specimen the same procedurewas repeated. Other weighings were taken in some cases, during as 248 PROCEEDINGS OF THE NATIONAL MUSEUM. well as after the first month, with always the same method of drainage,but without a change of solution.The method of draining steadily adhered to and applicable to specimens of all sizes, is to take the brain carefully into one or both hands,and then swing the arms with a somewhat rapid motion from forebackward, by which most of the liquid attached to the brain is thrown oti'; this takes only a brief time, after which the brain is placed forfive minutes upon a dry cotton towel. This procedure gives a goodand fairly uniform drainage, and is preferable to the use of funnels.In the second category of cases one of several additional proceduresintroduced was proportioning the quuntit}' of the preservative, incubic centimeters, to the weight of brains, in grams.The solutions chosen for the specimens here dealt with were 3 percent, 5 per cent, 10 per cent, and 15 per cent formalin (commercialsolution of formaldehyde) in distilled water; two solutions of for-malin, 5 per cent, to which was added salt, in one case enough to raisethe specific gravity to 1,035," and in the other to 1,030; and in addi-tion the writei' used several combinations of formalin with solutions ofordinary alum (potassium and aluminium sulphate), which was chosenfor its astringent effects on organic tissues. In two series a saturatedsolution of alum* was mixed with one part of water, and in anotherwith two parts of water. ?The changes to which most attention was paid, and which probablyrepresent best the physical changes, were, as with Donaldson, andFlatau, those of weight. The general and specific results follow:The changes in the weight of brains in all the mixtures showed(1) a characteristic typefor every solution, and (2) a noticeable variationfor every solution.(1) In every solution the first three to five days were with all brainsthe period of the most rapid changes in weight. In probably all ofthe solutions here reported upon, and with all brains, there was aninitial stage of gain. This reached more or less promptly its maxi-nuun. and was followed by a general, long-continuing loss. A periodof stal)ility was established but slowly. So far as the observationswent (two years), absolute stability in weight of the specimens was notreached. In every solution the daily changes in the brain weightformed a characteristic curve. This will be better illustrated inPart II.In all the simple solutions of formalin in water, up to 15 per cent ofthe former (the strongest tested), the initial gain was well marked.It was larger with the weakest solution and decreased as the propor- " Near the speeilic gravity of the whole brain; after Spitzka.f> A saturate.! sohuion of alum Avas prepared l)y placin-r an excess of that salt in alarjre jar of water, stirring' well, allowing the mixture to stand at ordinary tempera-ture for several days, and filtering just before using. N0.1451. BRAINS AND BRAIX PRESERVATIVES?IIRDLICKA. 249tion of formalin increased, which showed that the effe(;t of formalin onthe brain was to hinder its enlaro-enient (apparently due to wateralone) and probably, in addition, to promote the loss of some of theconstituents of the organ. After the culmination of the process ofgain, which, as shown bv further experiments, was always completedbefore nine days, a gradual progressive loss followed, which in per-centage was alike with the different solutions. The ultimate weightof the l)rain depends, in consequence of this similarity of loss, verylargel}^ on the height of the initial rise in weight. If this had beenlarge, as with the 3 per cent solution, the ultimate weight (within two3"ears) would still be above the original; but if the gain was smaller,as with the 15 per cent formalin solution, the ultimate weight of thespecimen would be found more or less below its weight imn)ediatelyafter extraction from the skull.Whenever a marked increase in the weight of the brain took place,there was also a noticeable increase in its volume.The eifect of adding alum or salt to formalin solutions was todecrease or, with larger quantities, almost entirely to do away with theinitial gain, and to augment the subsequent absolute loss of brain weight.The percentage of the loss in weight, however, remained related tothat in the simple formalin solutions, Simultaneousl}" with the loss ofweight in the stronger concentrations was also visible a decrease inthe volume of the organ. No direct relation was found betweenthese phenomena and the specific gravit}" of the solutions. It appearsthat alum, sodium chloride, and, according to more recent experiences,other salts also, as well as, alcohol, act on the brain physicall}' much likegreater proportions of formaldeh3"de; hence the use of such meanswith formalin permits the obtaining of similar physical results withcorrespondingly smaller proportions of this chemical.A renewal of the preservative generally affected slightly the changesin the brain, causing a temporary rise in weight.As to the rapidity of hardening and other visible changes in thebrain, the differences between the several liquids were not great. Amoderate toughening of the brain was in every one of the preserva-tives observable on the second day, and a good hardening, with freshadult specimens, was generally reached within a week. In the salt-formalin solution the brains were, at least for a time, slightly softer,in the alum-formalin solution slightl}" more resistant, than those pre-served in simple solutions of formalin of the same strength. A higher [ percentage of formalin was favorable to a more rapid and perhaps aslightl}' greater hardening. On the whole, should one be given speci-mens of the same size, but each hardened in a different solution ofthose here dealt with, after the}^ had lain a few months in the liquid,it would be quite impossible b}^ the hardening alone, as perceptiblethrough the unaided touch, to distinguish an}' of the preparations. 250 PROCEEDiyGS OF THE NATIONAL MUSEUM.The hardening of small mammal and bird brains was effected muchlike that of the larger specimens. In hardening fatal human brains,the best results were obtained h\ the aid of stronger alum solutions.The color of the brain (except so far as it ma}' be due to hemoglobin,which is bleached) was affected but little by any of the solutions em-ployed. Sodium chloride produced a lighter color or bleaching of thetissues; alum a slighth' grayish tinge of the surface. AUnn was moreeffective tiian salt in showing the differentiation of the gray andwhite matter.(2) ^^'ith all the care exercised, the ratio of change in an}' given seriesin which the same preservative had been used was not uniform. Alarge portion of the irregularity must be attributed to the physicalstatus, and some probably to the chemical condition of the organ. "When the two halves of any brain were treated in the same preserva-tive, the results were always much alike.The physical condition of the brain includes its size and the quan-tity of blood or other liquids it may contain. The size of the brain hasbeen found in general to have a pronounced influence upon the weightand vokune changes in the organ. The larger the brain, the smallerthe per gram changes, and the opposite. While there are individualexceptions, the cases conforming to the rule (see detail tables) are toonumerous to leave any doubt on this point. What the causes of thisphenomenon are is not yet clear, though presumably the larger brainshave a tirm(M- structure?that is, could better resist absorption"?andthe very small brains are of necessity preserved in relatively nuichlarger quantities of fluid, which may aid solution. It is possible thatit is mainly if not entirely the size which accounts for the differencesbetween the changes in three principal series of brains?those of humanbeings, of mammals, and of birds?but this needs further experimenta-tion before a flnal decision can ])e obtained.The degree of brain congestion nuist be a factor affecting the brainchanges, hut not enough specimens came to hand to throw much lighton this i)()int. Theoretically, a congested ])rain ought to gain less andlose more than a normal one. in any preservative. Higher degrees ofcongestion, not unconnuon in human specimens, are rare in otherlarger maninials and are practically never met with in the smalleranimals.Besides the differences in the changes of various brains in the samepreservative, accountable for l)y marked ditterences in the physicalcharacteristics of the organ, others are met with harder to explain.In some instances, as with Lepm ctmicultts, Cathartes aura, and a fewothers (see detailed lists), there is a suggestion that the difference "In a number of instances the hemispheres of small brains, preserved in weak for-malin solution (1 ur 2 per cent) in the laboratory, have burst through the greatabsorntion. BRAINS AND BRAIN PRESERVATIVES?HRDLWKA. 251may be that of species, which opens a large field of inquiry. But, inother instances, members of the same species, and that even whericollected and preserved under nuich the same conditions, show pro-nounced ditferences, and these can hardly be accounted for on otherbasis than chemical. The following- figures show two such instances,(1) in human and (*2) in bear's brains: X ^ j: 120 252 PROCEEDINGS OF THE NATIONAL MUSEUM. vol. xxx. r I ^ ^\ --1---"T>^4:^ ^.,,.4-.U - I BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 253BRIEF DETAILS CONCERNING THE VARIOUS PRESERVATIVES.THREE PER CENT FORMALIX SOLUTION.All specimens increased in weight, nianinial l)rains more thanhuman; brains of the ^'oung-, human and mammal, increased morethan those of adults. A decrease in weight in all classes of specimensset in within the tirst month and continued slowh' as far as observed(two 3'ears). In every instance the weight of the brain at the end oftwo years was still greater than the original. 254 PROCEEDINGS OF THE NATIONAL MUSEUM. VOL. XXX. I3= r- : { :?7 d- - / XT .2 ? NO. 1451. BBATNS AND BRAIN PEESERVA TIVES-HRDLICKA . 255 I i^ o 256 PROCEEDINGS OF THE NATIONAL MUSEUM. HnJliU ' i ! ii.i ; U.I Milfmm ^4 i 1 - -!!riMm.M^m -^?A-/ //iji; ,: i....j ,.i.TWV. M-i- i l- i'! ?-^U,...4^^.,.^.u.H-i4-t^ ! M i I ; I :/ [/Trnt' F 'r t 11 -K- i-4-U IN ! A 1 IEif i ! ?? --..I IriijMII IT i ttltt+rr NO. 1451. BBAIXS AND BRAIX PRESER VA TIVES?HRDLICKA . 257 i i 1 258 PROCEEDINGS OF THE NATIONAL MUSEUM. VOL. XXX.TEN PEK CENT AND 15 I>ER CENT FORMALIN SOLUTIONS.In these the number of mammals was rather small and all were ofsmall size, which had an effect on the figures. All the specimensincreased in weight, as in the 3 per cent and 5 per cent formalin solu-tions during the early part of the first month, and slowly and con-tinuously declined afterwards. The ultimate weight reached was ineyery case, and particularly in the 15 per cent liquid, lower than withthe weaker solutions. It was greater in the birds than in the mammals^Experiments on the larger and more uniform brains of sheep showedplainly a progressiyely less initial augmentation and lower subsequentfall in weight with the increase in strength of the formalin to 10 and15 per cent.l.im) AND 1,035 SPECIFIC GRAVITY SALT SOLUTIONS, WITH 5 PER CENT FORMALIN.These two solutions acted practically alike; the 1,030 sp. gr. liquidwas more largely used, for the reason that fewer brains will float in it. XXi- 90 per cent. l I .; 70 per cent. 4-4-^ , T-T"7ftMammals, adult, Bird.s, adultFig. 8.-CURVES showing changes in brain weights in 1,030 specific gravity salt formalikSOLVTION.Only adult mammal and bird brains were preseryed in the solution, nonormal human or 3'oung specimens haying reached the laboratorywhile it was being employed. A large majority of brains in bothseries showed at the end of the first week a decided loss in weight, BBAIXS AND BRATN PRESERVATIVES?HRDLICKA. 259and this gradually progressed. The bird brains showed greater lossthan those of niannnals. On the whole the effects of the solutionresemble those of the alum-formalin com})inations.ONE-HALF SATURATED SOLUTION OF ALUM, WITH O PER CENT FORMALIN.The various series of specimens in this solution behaved in the sameway as those in one-half saturated solution of alum with 10 per cent I I I I ? 80 percent \\-v- "\ Mammals, adult, Birds, adult Mammals, young, ? ? ? . ? . ? .Fig. 9. ? Curves .showing changes in brain weights in one-half saturated solution of alum,with 5 per cent formalin.formalin, except that the loss was throughout slightly less. Anincreased amount of formalin with this alum .solution favored some-what a loss of weight of the specimens. 260 PROCEEDINGS OF THE NATIONAL MUSEUM.ONE-HALF SATrRATEl) SOLUTION OF ALUM, WITH 10 PKR CENT FORMALIN.At the end of the tirst week one human specimen showed a smallincrease, another a slight diminution in weight; among the mammals,two adults and two 3'oung showed a very slight increase, the rest ofthe mammals and all the birds a decrease in the original weight.Apparentl}- there was an initial rise, but it was slight and of short 102 per cent,p- 90 per cent. 80 per cent. -- -?A "^ - -- 1 34i 70 per cent t Z? ..: IIL_. uAfammals, adult, ?Mammal.s, young, ? ?Fig. 10. Human, adult,Bird.s. adult,-Curves .sHowi.VG changes i\ bhain weights in one-half saturated sui.itiox of .\i.um,WITH 10 per cent formalin.duration. T\w hniins of adult l)irds lost more than those of full-grownmammals, and these lost more than the adult human ])rains. Brainsof young mammals lost more than those of full-grown. The ultimatedeticiency in weight Avas greater than in any of the simple formalinsolutions.ONE-THIKI) SATUKATKI) SOLUTION OF ALUM, WITH O PER CENT FORMALIN.This preservative influenced the weight in the various series ofspecimens much as did the one-half saturated alum solutions, only theloss of weiglit was on the whole still slightly smaller. The one adulthuman brain preserved in this liquid showed a slight initial increase,but in the mammal and ])ird brains there was at the end of a week in Ko. 1451. j^ji^uNS AND BRAIN PRESERVATIVES-HRDLICKA. 261most cases already a decided loss. The hv^Mu^or'ih^'^^^^^T^Yihuman and mammal, with one exception, lost more than those of theadults The brains of adult mammals and birds behaved generallymuch alike in this solution. ^Human, adult, Mammals, adult, Birds, adult, 110 percent. Human, young, Mammal,., young "?-"'~"""'??''"?""?^;?ri:r;?r""-""?"--"???--prIt.tfv'rr",f:f ""'^'' p''''" ""'"" ^^^ '??-'"' brainPC lalo e Tfti , ?,1'"'-'''"^'=t?' ??? '?'- ?ac,-o.seopie.l pur-poses al ne. It it is desired to preserve specimens of a mixed fhunianand comparative) collection near tlieir aetL] weight andTolumeTwo 262 PROCEEDINGS OF THE NATIONAL MUSEUM. vol. xxx.or three solutions of different concentration for brains of widely differ-ent mass would seem to be needed.Hijjhiy diluted (less than 5 per cent) and again hio-hl}' concentrated(over 15 per cent) formalin solutions, and large additions of salts, aredisadvantageous and ought not to be employed.Addition of alum to the formalin solution favors the process of hard-ening, and is to be reconnnended in preserving brains of the young,particularly of human fcetuses.Among the numerous points left to be determined are the effects ofadditional solutions, the influence of different quantities of the pre-servative, and the exact daih* changes during the first month at leastin the specimens. Experiments made in the laborator}^ during thepast summer with fifteen series of sheep brains and reported in PartII, will throw some light on these matters.The changes in individual brains of this first series are given in thefollowing tables: NO. 1451. BRAINS AND BRAIN PRESER VA TIVES?HRDLI^KA. rH inooeoeo ^ ooo P^S'I Pes bjo? fl'S 53 to63- ? iC lO O C^l iC' -.D lO OS CO OCU5 00 t^ Cs -f? C^l rH rH CC CI bog t.*- - 33 t. b : oi.^ OS c S S a; *^ uJu cj bjD CI -1^ be OS tuj O ^^ i-H CO QC C^CO -,C '^C^ 1-1 (M GOiCOOO COCl OCOCOOii-^ 'M* -'* CO 1-1 C4 rH ' CO" C^I rA I I I 1 + ++ +111 I + i IOOOGCO coco lOX'^r^ o 000 t^t^ao'OOiCOCCifl coos ?Dt^ id-^TfTjJ (N o-tp "^ococ^'coCOi-HrHC^.-H COIM C0r-(C^C^ ,-( ,-( -^ ^ S i5^ lOO O vCiC iC 00 "^O O O O O 00 i-H CO CO iC 'I' x" ^ r-" '^D (N GO ir: -^' ic oi o r>^ od ci r-I r^ t^Tcoco C^ i-H(N ClC^i-H "^ (MrH r^ i^ ift ci ic o ic ^' 30 t^^DCOrH^CO?-? CO-M COi-HC^iO l>C^00 cooc^i i-^ o ^'00000 00 iC o o oocic^i^co ai:o 1-40 ?co'^-riGOcoo oiQO rico'-cfn O iC Oi 00 00 csO '^ CO CO Tf 'iJ S : .0 ^< g : ? cS o . ? OJ'O ??^"^ : too o CO CO cc CO CO coco CO CO CO CO CO coco00000 00 0000 o 00t^ a; rt X X CO o??( C^ i-t CJ C^ CO <1"0 J3 X r- -^ '? ? 5 o- 05 00 00 xi 01 oi r^ i^ JM o ^ iCi?I 00 Ol O tHcocococooo CO o^ -r CO CO CO -J, ^ ^ ^ ^ -r ^ .---.-C^ C^l 01 C-1 2f)6 PROCEEDINGS OF THE XA TIOXAL MUSEUM. ' t: .'*'= St iP* .-I ?- t^ x" ci ci T" 3D C'l ??' C-l I^ C-J ^a> 3i t5> OS 00 0> S5 OS Oi OS QC Oi X 05 X 'tis '-~ i ;o cc ^^ OS OS '5 ^ I^ '.^' ?-? ?*!;5 O OS -.O iC O iC ?2l^ CO C-l C-l X u- o^ OXi^f-H-OC-1f^ '.Sfaso t, o he j; = X = t.^" ^s=sa I +OO COiO OS t^ ' '-difi'CC ? - .-iC-lf-H COM^t-HC^ O f-HCO O OS cc ?-?lO O -w O OSD O X 1^ -^ ^* lit C^COXX i^Xi/tXCO CO irt CJOS O ?5 rt O =-? .-I -?S-H . Xr-lX "^i^ cc 00 ^" ri ' r- CICC CCgCSXCO XM ?C OS OS *-? ?C 5 OS e-. c-- c- ^ c bp..^ S-r^ C3 K -..i 5 oso t^ o -r OS5j m' x' o c4 -t t^ ? CO CC CO CO cc cccocccooooooo ooooOSOSOSCSOiOS CSOSOSOSOS t^ X OS CO ^H O t?t -I* o O cS o *J*J*J > > >U C> o o o c > > > i _O O O O ? O zj c o c .13 ^ Q S ^? ?, oE 9 .a r c 3 ^ 2.Sc u u ^ sx: O >s o 5.3 = 3. = _^.g|l fe -<2?^tr''^'-^*^ ocotcc^^oscoJScoco ? ^-T -T-r^^-^cccoJl ~1 M Tl M M M r) M ;.l ^.1 M -I 'c;li'E QJo "oco BRAINS AND BRAIN PRESERVATIVES?IIRDLICKA. 267 "<-.? tie Ceo -0 a! Q2-3 "-5 tie - ^5 . , i_i j^ C t^ >i( , o cs? aj^^-c o CD03 o ^3=* a 1^ t-H iH t^ IC OO lO ?C ^ OS oCO c? 00 t^ t^ 'X ?-*a3 00Q0=O Ol>0:00 "l^CCCOCCiOc-i oi o ^' 00 _ -X t^ X 00 X ? I~ X X X X Oif-i -.c :0 O00 r-i IC CC rf" C-l i-H ,H . c-- ic 1-H oi C-. CO C- C-- 00M ! ! I I CO C^ c^. OS iCOi-H oo 05''^cc>-^ooiccocoiCocO'MTj'coiCTPTj?r^cv:iu:5Q'^?-HOGCai'T'o6lC3CGOCOi-H--lJr-HTJ^rH-1':DI?lOTOsOOasOiOiOOOOiOC^Oi-HOOOO C "^ C> c^- CO '^6 ^ OOCCi?tt^COOS tDQOiCQCi-O-f.?'CO C* rH i-I r-I M O t-^ t^' 1-i X CO C^i iC t^ - s. f>. 00 -r^- *- COiC. r-I c4 - <;O00GO rH? 1^' Oi GC C-. t^V- ' lO lO CO !>? Ol iC "^a5 -OOOrHt^OS?D(M(MrH'^.-HC^*C^ca(:c?OrHac(N^cocoTr'Tl^o a . : : :3 O O O O 3 M a, 3^ ?'? OS I 3 *-o^ S-^ c I 3 u? ' -""OJ 3 C ?-1 - ?-"ui cS "CI C '3 o ^a ^'ss-a "3 2,^S ^-^ =S' ' J= .???"? ?I'u rt ci i.S " 3'S5533?^ ct ^ ? ^O ospa -O lO -f ? cCO^'-HlOrH lOTCOOl-f O .-^ -O O CO X ?C 'M OS O 'M r^ c-i cC^ C-1 ;?) IM CM _ _ _ . 4 -r 01 ri CO c>j rf Soooo oooooooooooooooooooo ^3'0'0'0'0 'O 'O 'O 'O 'C 'O 'CO 'CO 'O 'O 'CO "0 'O 'CC'C'C ?"o ? ? ? ! I : ; 1 ; ; S< \ ; \ :::;:;::??:::: i ;;; i ; ? -t*-fcO'^'^ -t'-r-fcoco-r'^-fco-r-f-f-fcC'COco-f*t"-j'-^ -^ooooo oooooooooooooooooooo o ^ c^ oo''30 rf oTlO lO ',o o (M ic (oTco t-Tos c-fco (rrj->-'":o'^rc4^oTir:r im'j? I?' cJ tH r" t^ ^ ^ c^ > ri ri ^ C; ri r- '-* .D d > (li (?' ^ ^ ti ?-' i C^l -M (M C^I C"4 Ol C^ (M (M M (M CJ C^I (M n M C-1 C^ (M C^(M C^l ? ^ ro cc00 ai ^ -^ "^t^ (M* c^ lo oi:o r^ ao i:^i> I-^-OI> nC0OrHCniCC(NX'-H-I'O<;CC^C0'7<'*CC .-H CO lO OC c4 C^i i-H o6 CO t-^ Tp (N x' C^ era iC CD ' C^' I>COri -( 1-1 T-H t-HtH (N, I I 4- [ M I M I I I I I I M I I I rH-t"^ iMOoC'i-ti-HOitM-O'-HtOiM'?lOt^X^O^CO " I> iC 00 OC - c^iOi?'Oi-roccGCior?fi? t iXit^X'OoocaiCit^aiOioooiooaiOif^c ' ?C IC lis lO(N *rHr-4t>^Oa5COCCiO-^COr-irHO'<^*Tjic OOO OOOOOOOOOOOOOOOOOOOj'ri>icicO'--c^t^cotoc50^HcooO"^-rxMi-H C^O^Jrti-lrH' OH ?r ''^ioecco?:)(MOo^ o o oi as o o : ^ O iC iC COooooco ^aOGO-r-pxxaoxooaooocoxaOGOccGCXN tN (N C^I CS CM C^l C-l O) C^l C^ C^ C) C^ OJ !N CM Ol C^ OJ C-1 CICMfNN C^dCMCMCM-lCMCMCMlMC^CMCMIMCMCMCMCMCM CCg y-5 ES SpqOOOcot-OS o^ OS OS ci '-' 270 PROCEEDINGS OF THE NATIONAL MUSEUM. "^ Isa. ;3 t^OD C^ C^ 3^OC f.^ lO Oa ^iC ts OC iC?OC X X X o> . O ? lit O 7-1 : 1-! si in --c o: CO c^ U-: ? -T bcgJ^'S--- .<" s - - -u 5 jj ? -^ S -o B - o oS^n "Si- 5|z ^ CC -^ iC X r- xa5 X i-H ?O u- O O ?! toic lit x" ro r^ X*C> 01 wl Oa 35 X . CO ic ir: o XS -^ m' '-' oi -f^ c-i Oi00QC00GOl>l^C3CGOQ0 oo t^Tj* .-( r OO ?" i; a> c ceo CN O COrr C^ r-i iO(N coco(N "^ r-li-l . '^ , oj 3 0) s L ?01 '-^ OJ; "Tfco'.oiOTPc^iiocooas'-Hl^C^IC0r-IC^?COC00i0s:07 i I I 1 1 77 I I I C^ O l^ c-?.COt^O CJ I 1 7 I -3.92 -8.61 -7.35 -13.65 -10.40 -10.26 272 PROCEEDINGS OF THE NATIONAL MUSEUM. VOL. XXX. t. c c-r ^i"^! OiC CO CO OS O iC t; * * T-H i-H i-H .-H t^ O iC '^ t-^ O 1 cc CI C5 aoiC COCi CO 1-H .-J 00 ^ C^ (D -^ ^ vOX l> C>- X GC o t. c ?- __ .c r* GO CO o 00 trtCCOO CO oio to 1 + I I 7 I O -*O O 'J' * 02 IM rJ rt Ol l> to tH ? CO Ol (N t^ iifliO>c tc c-ioitoiOt>Q6o6coi/soX'^iCcotdiiioioo't>toio-j Ic-i CO O-^M-^ XOOOi 050300 X 00 -oif-io^<-tf--os'^.-HTrxi^cN0030iictoa>c-i ^ ^ r-^ ^ CO oi ifr oi 00 o ic ->! r^ 1--^ OS oc" ic ic c4 -I- " X01XX01!X'01QOOOXCn00100?OOOCXOOOSC-. c-- ^.---.x CCCCX-^t^OiCOi-i'^OT-'i-'CO'^OS'^OStOlCOiXO^1^ uc to x' T-* ic ^* CO co' -^ 1-H T-i rH to X* ci c^* CO CO c4 ci : ' c^ r^ -r " t- P 'T*,aj 5 - o OCO iC Oi^lC M-^COOiCl-TJ'CSCO^Ht^OINXtOOiC'^XiCC^XtO-5'OXr--.o ? ?rH^H : : : :OQOOOOOS OS OS OS OS OS oo'co"co"c^rc4't>^t-H Ol (M rt l-H a; -^ 0) O CJ 3ai-?!a300-C45 iCCOiCOOO'^OOO? 00 to ^O I> ' ^' .-^ Olt-OOii-HCJiCCO c^ Oi c^a>io'^ i-^ ci * i-H o^t-txt^xcoco-^ ?-iocooo^or-r^co*-D oicoiC'tOiTTCO'^Olt^'MrHrH OiiOrOOOrHOOCOCOi-H t^i- r4 O ICQOOOOS ^ 53 ? '^ ^ ^ C -r o OS CJ ri ?r:-*'t^C0l-O:0t0Ot^O OiCCJ-^X ci * r-i " i-H * u': x ci i'^ ?c oi OS OS a : .:::.: : : .30 ooooooooo ooocoooocco o?^oo;S'C 'O 'C'O 'O 'O 'C 'O 'V'C 'C '^ 'O "CO 'C'C'C 'C'O 'O 'O'O'O'O oocoocooo oooocoooooo cs>. ooOOC^OiOOO OOO 'OOOOOOO > c- r^ c- c^ ^. Oi ^1-1-^ ;Or i cr.cn ;""jToo'ci'^'eoiM" oco'io" 'CiOiOiOSGlS^Gi C3C3N r-IMo a>550 o o o o u J C^ rH r) rir-( C-) ?td <0 O 'r-l 00 TjlCJ to CO O ?5 oa>a> *-? CO -^ C^ C^ r- '^ COQOO t^ oc CO toc^ oo ooootoio^OtOtOOfN CO'J'I^ifltncOXOOOaa CO t^ O^ CD 0000 OS 00 00 00 Oi 05 OS 00 Oi t>- Ol O 0> 00 CC 00 OS GCOil^QOOO cec^cooooioxosSc>id-j>r~cod?iS in r< 14 l^ C-)5 r-icooo-^-.cicoiirac~r-OJOt^COHiOOO^'^r-looTJ 'co'od>-icodci Oi CO 1^ ?o o.__ _ Oiirjt--^c^inrH 1? i-H i-H f-H C^TTrHCOOO to Oi ri Oi 1^too coos TfCO O 1^ T- '^ O c- ^l ^"^ to I> to 00Oi^Oi Oi OOiOO''Oi 3 r- -.. ts oi ,^. e.. - o i-H i~- c-1 n r- COOS ri ?.. ^. to ,? ^. -r ^ ~i 1-c ic tc t^ 1^0Ct1< Os^ torH OC COTf ,-. ^'O 2 o 1.0 to rt o o rt2 'C ^ oi OS -ji X tjiw to ri -M r>. n x' m' ' Tj- 1--' d m' ^' ?-? oi O X CO'^iC oid iC to c^* t>^ oi>? co-*c^coo S ?s 5 * 5 . o in .? >. ? u, C ^ > ?^ < < S S "^ >^ to'oTic c^x'df-H'os ri to -S" -)> -f -* 'i"o ooooOS Oi Oi OS OSC^ r-^ CO i~ to c^ -r to :i-i 1-1 1-1 rH C^ C^ -- _ o j: o =: ^ 7 - s ja^ll S .. i) C 3s Oi - : a00> 'OfX CO OS rr TT ^N ?-? n to to 1^ I^ to ^^ ^I Ci OS3r i? S 2 S S 3" in ^ icic CO -s" ic CO -T ^ --?222?S cjiot^ooioiosoxo^ liIN M ^) c^ ?) c^ fi c^ 6J S jj ?S 53 6) ?J jq ?i O (M to O >Cn o OS to oOS X 1^ ? X^ -r Tj. X -r.-< ?^ C^ C^l C-l ?^?J I^CMC^C40) PART II.PHYSICAL CHANGES IN SHEEP BRAINS COLLECTED AND PRE-SERVED UNDER SIMILAR CONDITIONS IN VARIOUS FORMALINPREPARATIONS.The first part of this paper dealt with the effects of formalin pre-servatives on the weight of human, other mammal, and bird brains,adult as well as young, collected under various conditions. The resultsin any solution, although more or less characteristic for that particu-lar liquid, were by no means uniform. It was found that, in general,the changes in the large brains were less than in the small ones, andthose in the adult less than in the young. Some of the differencesmay eventually be found to be those of species or larger subdivisionsof the animal kingdom; but be3^ond all this there was seen a consider-able and unaccountable variation. This element was recognized longbefore the first experiments were completed. It rendered desirablea separate series of observations on the brains of some fair-sized animal,collected equally fresh, extracted and subsequently treated in the samemanner, and kept in proportionately the same quantities of the preserv-ative. Under such conditions the action of the various preservativesshould be much clearer and more comparable, and the differences inthe changes be reduced to the minimum ; if noticeable disagreementsstill existed, they would point to differences in the structure of thebrains or in their chemical composition.It became possible to undertake the series of experiments duringthe earl}" part of the summer just past. An arrangement was madewith one of the city butchers to deliver ever}" morning a small seriesof heads of sheep killed the night preceding. The brains, with thehelp of the laboratory aid, Mr. Docekal, were extracted in as short atime as possible and in the same manner (see Part I), then weighed andplaced in a proportioned quantity of the preservative. Fifteen serieswere determined upon and the specimens were secured in a little overtwo weeks, during quite uniformly warm weather. Every seriesexcept two, which were smaller, consisted of eleven brains. Ten ofthe specimens were placed in a quantity of the preserative amount-ing to 3 c. c, for each gram weight of the specimen, while with onethe quantity to the gram was made 6 c. c. Of the brains in 3 c. c. tothe gram liquid, one of about average dimension was weighed everyday the same hour, while the remaining nine and also the eleventhspecimen were weighed at the end of seven and again at the end of277 278 PROCEEDINGS OF THE NATIONAL MUSEUM.thirty days. Other weighings, as indicated in the detail tables, weretaken thereafter. At the end of the seventh and the thirtieth day thesolution and cotton were changed, as is done with all specimens in themuseum collection. The drainage of each specimen was as uniformas practicable by the method outlined in the first part of this paper.The solutions employed were:1, Three per cent formalin.2. Five per cent formalin.8. Ten per cent formalin.4. Fifteen per cent formalin.5. Saturated solution of common salt with 5 per cent formalin.6. 1,030 sp. gr. common salt solution with 5 per cent formalin.7. 1,015 sp. gr. common salt solution with 5 per cent formalin.8. Saturated solution of alum with 5 per cent formalin.9. One-third saturated solution of alum with 5 per cent formalin.10. One-fifth saturated solution of alum with 5 per cent formalin.11. One-third saturated solution of ahim with common salt up to sp.gr. 1,030, with 10 per cent formalin.12. Saturated solution of alum with 5 per cent formalin.13. Eight}' parts of 1>5 per cent alcohol and 20 parts of 5 per centformalin.14. Sixt^'-five parts of 95 per cent alcohol and 35 parts of 3 per centformalin (near that of Parker & Floyd).15. Sodium acetate 130 grams, sodium chloride 110 grams, formalin20 c. c.,95 per cent, alcohol 460 c. c, water 540 c. c. (Stroud, Wilder).The results, in detail, were as follows:THREE PER CENT FORMALIN.End of first week: The weight of brains in the 3 c. c. to the gramsolution had risen in average 21 per cent, or over one-fifth of theoriginal. Variation: From 118.55 per cent (specimen of 102 gramsoriginal weight) to 123.9 per cent (specimen of 102.5 grams)= 5.37per cent. The two heaviest brains (117.5 and 110.5 grams) gained,resi)ectively, 20 uiid IS. 55 per cent in weight, the two lightest ones(94 and 93.2 grams) 20.2 and 22.1 per cent. The brain in the 6 c. c.to the gram solution (100.5 grams original weight) increased 18.9per cent, less than any other except one of the heaviest specimens,and the one weighed every day, which may have been affected thereby.End of first month: Weight of five of the nine brains in the 3 c. c.to the gram solution is very slightly greater (+0.08 to +0.44 percent); of three, slightly smaller (-0.42 to -1.15 per cent), and ofone, equal. Variation: From 117.19 to 124.(12 per cent^'=^6.8 per cent.The changes were not quite harmonious with those of the first week,or proportionate to the weight of the specimens. The brain in the 6 ? As compared with oriyrinal weight. NO. 1451. BRAINS AND BRAIN PRESER 1 'J TIVES?IIRDLICKA . 279 r 280 PROCEEDINGS OF THE NATIONAL MUSEUM. c. c. to the gram solution lost most and is now relatively the lightestof the whole series, excepting the one weighed every day.The brain weighed daily showed a great gain during the first day,reached maximum on the fifth, and began to decline on the seventh;after change of solution it rose during two days, and then again beganto lose, which was repeated identically after the one-month change.FIVE PER CENT FORMALIN.End of first week: Weight of brains in 3 c. c. to the gram solutionhad risen in average 17.9 per cent, or a little over one-sixth of theoriginal. Variation: From 110.12 (specimen of 119.7 grams originalweight) to 120 per cent (specimen of 99 grams original weight)= 3. 88per cent. The two heaviest brains (119.7 and 117 grams) gained,respectively, 16.1 and 18.4 per cent in weight, the two lightest ones(98.5 and 95.5 grams) 17.8 and 19.1: per cent. The brain in the 6 c. c.to the gram solution (100.5 grams original weight) had risen 18.9 percent; that weighed every day 17.8 per cent.End of first month: Onl}' one specimen showed a slight gain (-f1.08per cent), while in eight there was a loss (?0.86 to ?2.39 per cent).Variation: From 113.87 to 119.66 per cent=5.79 per cent. Thechanges were not quite harmonious with those of the first week, orproportionate to the weight of the specimen. The brain in the 6 c. c.to the gram solution lost 2.09 per cent, that weighed dailv 2.39 percent of weight, both above the average.The brain weighed daily gained much on the first day, reached amaximum on the fourth day. declined slowl}^ to seventh, rose afterchange of solution during two days, fell gradually' to the end of thefirst month, then, after a change of solution, rose one day and has beenslowly losing since. TEN PER CENT FORMALIN.End of first week: Weight of brains in 3 c. c. to the gram solutionhad risen in average 15 per cent, or nearly one-seventh of the original.Variation: From 112.84 (original weight, 97.5 grams) to 116.87 per cent(oi-iginal weight, 80 grams)=4.03 per cent. The two heaviest brains(1 12 and 110.5 grams) gained, respectively, 11.73 and 15.38 per cent,the two lightest (96.5 and 80 grams) 15.54 and 16.87 per cent. Thebrain in the 6 c. c. to the gram solution (125 grams original weight)gained but 14 per cent of weight, that weighed daily (102 grams orig-inal weight) 16.17 per cent.End of first month: The weight of one of the nine specimens in 3c. c. to the gram solution has very slightly (+0.45 per cent) increased,that of the other eight slightly to moderately (?1.07 to ?3.29 percent) decreased. Variation: From 110.31 to 115.63 per cent= 5.32per cent. The changes did not quite harmonize with those of the first BRAINS AND BRAIN I'RESERVATIVES-HRDLIVKA. 281 282 PROCEEDISGS OF THE NATIONAL MUSEUM. NO. 1451. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 283week nor were they proportionate to the weight of the specimens.The brain in the 6 c. c. to the gram solution lost 1.05 per cent inweight, that weighed daily 3.38 per cent.Changes in the brain weighed ever}- day: Considerable rise the tirsttwent^^-four hours, continuation of rise until the sixth daj^, then slowdecline; a moderate rise of two days' duration after the first and ofone day after the second change of solution.FIFTEEN PER CENT FORMALIN.End of iirst week: Weight of specimens in 3 c. c. to the gram solutionhad risen in average nearly 13 per cent, or one-eighth of the original.Variation: From 107.61 (original weight, 98.5 grams) to 116.48 percent (original weight, 91 grams)= 8. 87 per cent. The two heaviestbrains (119 and 111.8 grams) gained, respectively, 12.1 and 13.5 percent, the two lightest ones (94 and 91 grams) 11.7 and 16.4 per cent.The brain in the 6 c. c. to the gram liquid (original weight, 97 grams)gained but 9.08 per cent, less than any of the above with one excep-tion, that weighed daily (original weight, 105.5 grams) gained but 8. 53per cent.End of tirst month: Weight of all the nine specimens in 3 c. c. tothe gram solution has diminished (?0.88 to 2.31 per cent). Variation:From 105.58 to 115.38 per cent=9.8 per cent. The changes arefairly harmonious with those of the first week, but are not propor-tionate to the weight of the specimens. The brain in the 6 c. c. to thegram solution lost 5.95 per cent, much more than any of the above,that weighed dail}" 3.49 per cent, also more than any of those in similarquantity of solution but weighed less frequently.Changes in the brain weighed daily: A moderate rise in weightduring the first twenty-four hours, maximum of rise on fourth day,then slow, continuing loss; one daj^'s rise after each change of solution.SATURATED SOLUTION OF SODIUM CHLORIDE, WITH ,5 PER CENTFORMALIN.End of first week: Weight of l)rains in the 3 c. c. to the gram solu-tion had diminished in average by 7 per cent, or one-fourteenth of theoriginal. Variation: From 90.33 (original weight, 95.2 grams) to95.19 per cent (original weight, 104 grams)= 4. 86 per cent. The twoheaviest brains (107.7 and 105 grams) lost, respectively, 6.6 and 5.72per cent; the two lightest (89.7 and 84.5 grams) 5.8 and 5.9 per cent.The specimen in the 6 c. c. to the gram solution (original weight, 95.5grams) lost 6.8 per cent, that weighed every day 7.9 per cent.End of first month: Weight of all nine brains in the 3 c. c. to thegram solution decreased (?3.15 to 5.23 per cent). Variation: From85.61 to 91.82 per cent=6.2 per cent. The decrease was quite alikein most of the nine specimens and harmonized somewhat with that of 284 PROCEEDINGS OF THE XA TIOXAL MUSEUM. ? = NO. 1451. BRAIXS AND BRAIN PRESERVATIVES?HRDLICKA. 285the tirst week, but was not proportionate to the original weight. Thebrain in the 6 c. c. to the gram solution fell to 90.05 per cent of origi-nal weight; after which it diminished imtil it became relatively thelightest of all; that weighed every day was damaged and discarded atfirst change.1,080 SPECIFIC GRAVITY SODIUM CHLORIDE SOLUTION WITH 5 PERCENT FORMALIN.End of first week: Two of the nine brains in the 3 c. c. to the gramsolution showed a slight increase, seven a slight decrease, in weight.Variation: From U3.86 (original weight, 106 grams) to 104.2 per cent(original weight, 107 grams) = 10. 3-1 per cent. The two heaviest brains(112 and lOT grams) gained, respectively, 2.68 and 1.2 per cent, the twolightest (95 and 94 grams) lost 1.58 and 3.73 per cent in weight. Thebrain in the 6 c. c. to the gram solution (original weight, 105 grams)lost 5.72 per cent, more than any of the above but one, and thatweighed every day (original weight, 103 grams), lost 2.92 per cent inweight.End of first month: All the brains in the 3 c. c. to the gram solu-tion lost slightly in weight after the end of the first week (?1.3 to ? 3.72 per cent), but two are still slightly heavier than originally.Variation: From 92.45 to 102.33 per cent=9.88 per cent. The changeswere not wholly harmonious with those during the first week, or pro-portionate to the weight of the specimens. The brain in the 6 c. c. tothe gram solution lost more than an}" but two of the above (2.02 percent) and is now relatively the lightest; that weighed every day lost 3per cent and is now also among the relatively lightest specimens.Changes in the brain weighed daily: The first day a very slight lossof weight, which continued slowly till the first change (the first dayafter which there was an insignificant gain) and then up to the onemonth change (which produced no result). After the thirty -fifth daythere were two weeks of stability, with a slight loss following. 1,015 SPECIFIC GRAVITY SODIUM CHLORIDE SOLUTION WITH 5 PERCENT FORMALIN.End of first week: Weight of brains in 3 c. c. to the gram solutionhad risen in average a little less than 2 per cent, or a little less thanone-fiftieth of the original. Variation: From 101.29 (original weight,116 grams) to 102.45 per cent (original weight, 102 grams) =1.2 percent. The brain in 6 c. c. to the gram solution, (123 grams originalweight) rose 2.03 per cent; that weighed daily (11.4 grams originalweight) 1.31 per cent.End of first month: Weight of both specimens in 3 c. c. to the gramsolution has diminished (?1.43 per cent and ?1.7 per cent). Varia- i tion=1.41 per cent. The specimen in 6 c.c. to the gram solution 286 I'ROCEEDIXGS OF THE XATIOXAL Mi'.'SEUM. vol. xxx. BHAim AND BI{AIX PRESERVATIVES?HRDLICKA. 287 288 PROVEEDIXfrS OF THE NATIONAL MUSEUM.(original weij^ht, 123 grams) lost 2.39 per cent in weight, more thaneither of the above; while that weighed daily (original weight, 114grams) lost even more, or exactly 3.03 per cent.The brain weighed daily fell very slightly in weight during the tirstday, rose slightly during the next two days (reaching maximum thethird day), remained stationary the fourth day and then began to lose.It rose the first dr.}' after a change of solution, remained one day sta-tionary, and then lost slightly, gained again a little, and then con-tinued to lose to the end of the tirst month. No rise followed the onemonth change, the specimen remaining stationary in weight for oneday, and then went on losing.SATURATED SOLUTION OF ALUM WITH 5 PER CENT FORMALIN.End of tirst week: Weight of brains in 3 c. c. to the gram solution hadfallen in average nearly 23 per cent, or nearly one-fourth of the original.Variation: From 74.27 (original weight 103 grams) to 81.74 percent(original weight 115 grams) =7.47 per cent. The two heaviest brains(115 and 109.7 grams) lost, respectively, 18.26 and 23.25 per cent inweight, the two lightest ones (92 and 88 grams) 21.74 and 23.87 percent. The brain in the 6 c. c. to the gram solution (original weight125.2 grams), the largest specimen in the series, lost but 17.74 percent, hence less than any other; that weighed ever}' day (originalweight 101 grams) lost 25.75 per cent, which is more than any of theremaining nine in same solution.End of one month: All of the ))rains in the 3 c. c. to the gramliquid had suffered noticeably further loss, and that from 4.86 to 9.04per cent. Variation: From 61.08 to 74.45 per cent =10.37 per cent.The changes were not harmonious with those of the first week norproportionate to the weight of the specioiens. The brain in the 6 c. c.to the gram solution, though large, lost considerable (10.68 per cent);and that weighed every day became relatively lightest but one of allthose in the 3 c. c. to the gram solution.The brain weighed every day showed a great loss during the firsttwenty-four hours, lost slowly and steadily for eighteen days, remainednearly stjitionary during next thirty days, and then lost slightly again.The (lay after each change of solution an insignificant rise took place.ONE-THIRD SATURATED SOLUTION OF ALUM WITH 5 PER CENTFORMALIN.End of Hrst week: Four of the nine brains in the 3 c. c. to the gramsolution showed a very slight increase (+0.42 to +0.90 per cent), fivea slight decrease (? 1.43 to ?2.59 per cent). Variation: From 97.41(original weight 116 grams) to 100.9 per cent (original weight 110grams) =3.49 per cent. The two heaviest brains (120 and 116.5 grams)showed, respectively, 100.42 and 97.85 per cent; the two lightest ones NO. 1451. BRAINS AND BRAIN PRESER VA TIVES-HRDLICKA. 289 Proc. N. M. vol. XXX?06 19 290 PROCEEDINGS OF THE NATIONAL MUSEUM. vol. xxx. NO. 1451. BRAINS AND BRAIN PRESER VA TIVES-HRDLICKA. 291K)3 and 9< grams), 98.54 and 100.51 per cent of their original weight(Ihere IS no relation apparent at this stage between the changes andweight ot the specimens, but ultimately the originally heaviest brainshowed with one exception only, the least loss, the lightest brain thegrea es loss. The specimen in the 6 c. c. to the gram solution (originalweight 122 gi-ams) lost 2.05 per cent, with two slight exceptions, morethan any of the above; that weighed daily (original weight 108 grams)losmgl.l< percent, or more than any other specimen in the wholeseriesEnd of iirst month: The weight of the specimens in 3 c. c to thegram solution had diminished from 1.15 to 11.27 per cent (the heaviestbrain losing least, ih^ lightest most). Variation: From 89 17 to 96 25fr" '"'Ir?" ^''' '""*? ^^" '*^^"^^^? ^^^'^ ??<^ harmonious withthose ot the first week nor proportionate to the original weight of thebrains. The specimen in the 6 c. c. to the gram solution lost 5.44 percent more than the majority of the above, that weighed daily 11 11per cent more than any but one in the whole seriesChang-es in the brain weighed every day: A slight increase inweight the first twenty-four hours, followed by gradual, steadydecrease, apparently not affected by either of the changes of solution.ONE-FIFTH SATURATED SOLUTION OF ALUM WITH 5 PER CENTFORMALIN.End of first week: Weight of one of the nine brains in the 3 c. c tothe gram solution had very slightly decreased, of one it was the same asoriginal, and It had slightly increased (0. 43 to 3. 12 per cent) with the sevenremaining. Vanation: From 99.09 (original weight 109.5 grams) to103.12 per cent (original weight 96 grams) =4.03 per cent. The twoheaviest bi^ms (116 and 115 grams) had gained in weight, respectively,0.43 and 0.8v per cent, the two lightest (99 and 96 grams) 3.03 and 3 12per cent. The brain in the 6 c. c. to the gram solution (original weight126 grams) had lost 1.59 per cent, or more than any of the above, while 1 that weighed daily (original weight 100.2 grams) gained 1.99 pe; cent. ^ l.nd of first month: All of the nine brains of the first group had lostI in weight (7.35 to 12.83 per cent). Variation: From 87.56 to 95 45 I per cent =7.89 per cent. The changes, while not differing greatly , were not harmonious with those of the first week nor proportionate ^ to the weight of the specimens. The brain in the 6 c. c. to the gram ' solution lost in weight 7.66 per cent, that weighed dailv 9.76 per centor more than any other in the whole series.Changes in the brain weighed daily: A moderate increase, reachingmaximum on the third day, and then a slow continuous loss, notaflected by the changes of solution. 292 PROCEEDINGS OF THE NATIONAL MUSEUM. NO. 1451. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 293 294 PROCEEDINGS OF THE NATIONAL MUSEUM.ONE-THIRD SATURATED SOLUTION OF ALUM WITH SODIUM CHLORIDETO 1,030 SPECIFIC GRAVITY, AND 10 PER CENT FORMALIN.End of tirst week: The brains in the group of nine in the 3 c.c. tothe gram solution all lost moderateh' in weight (? 2.99 to ?9.74 percent). Variation: From 90.20 (original weight 113 grams) to 97.01per cent (original weight 100.5 grams) =6.75 per cent. The twoheaviest brains (115.2 and 113 grams) lost in weight, respectivelj-, 7.99and 9.74 per cent, the two lightest (100.5 and 95.5 grams) 2.99 and5.24 per cent. Ultimately, however, the heaviest brain shows theleast loss, while that of the lightest specimen is among the greatestlosses. The specimen in the 6 c. c. to the gram solution (originalweight 115.5 grams) lost G.OG per cent; that weighed daily, however(original weight 105 grams), 10.95 per cent, or more than an}^ otherin the series.End of the tirst month: The brains in the 3 c. c. to the gram solutionall show a further marked loss of weight (?6.6 to ?13.81 per cent);tlie heaviest ))rain had lost the least, the lightest the most. Variation:From 78.70 to 85.94 per cent =7.24 per cent. Changes were not har-monious with those of the first week, and were more in a reverse thana direct proportion to the original weight of the specimens. Thel)rain in the 6 c. c. to the gram solution lost 11.52 per cent, that weigheddaily 11.76 per cent.Changes in the brain weighed daily: A moderate loss the lirst andsecond day and gradual loss, unatiected by the changes of solution,thence onward.ONE-THIRD SATURATED SOLUTION OF ALUM WITH SODIUM CHLORIDETO 1,080 SPECIFIC GRAVITY, AND 5 PER CENT FORMALIN.End of first week: Weight of brains in the 3 c. c. to the gram solu-tion had risen slightly (+0.44 to +4.08 per cent). Variation: From100.44 (original weight 114 grams) to 104.08 per cent (original weight98 grams) =3.64 per cent. The two heaviest brains (123 and 114grams) had gained, respectively, 4.06 and 0.44 per cent, the two lightest(98 and 97.5 grams) 4.08 and 3.58 per cent. The brain in the 6 c. c. tothe gram solution lost 1.66 per cent, that weighed daily 4.63 per cent.End of tirst month: Weight of all brains had notably diminished(? 13.67 to ?21.12 per cent in the group of nine). Variation: From81.72 to 89.84 per cent =8.12 per cent. The changes were not pro-portionate to the weight of the specimens. The biain in the 6 c. c. tothe gram solution lost in weight 10.13 per cent, less than any of theabove, and that weighed daily 9.22 per cent, or still less than the pre-ceding, which compensated with both specimens for the loss duringthe first week.Changes in the brain weighed daily: A slow loss from the first dayonward, aecelerated slightly the day after each change of solution. NO. 1451. BRAINS AND BJiAIN PRESERVATIVES?HRDLIVKA. 295 296 PROCEEDINGS OF THE NATIONAL MUSEUM. NO. 1451. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 297EIGHTY PARTS OF 95 PER CENT ALCOHOL AND 20 PARTS 5 PER CENTSOLUTION OF FORMALIN.(Alcohol, 80; water, 19; formalin, 1.)End of first week: All the brains in the 3 c. c. to the gram solutionhad lost in weight; the average loss was 11.5 per cent, or one-ninth ofthe original. Variation: From 85.27 (original weight 112 grams) to91.45 per cent (original weight 117 grams). The heaviest brain lostleast. The specimen in the 6 c. c. to the gram solution (originalweight 106 grams) lost more than any of the above (15.1 per cent),that weighed daily (original weight 109 grams) lost 11.47 per cent.End of one month: The solution was not changed at the end of thefirst week nor at the end of the first month, except with the specimenweighed daily. No especial difierence appeared in the results. Allthe brains in the 3 c. c. to the gram solution lost slightly in weightafter the end of the first week (?0.77 to ?2.09 per cent). Variation:P'rom 83.48 to 89.31 per cent =5.83 per cent. The changes were notharmonious with those of the first week nor proportionate to theweight of the brains. The specimen in the 6 c. c. to the gram solutionlost 1.66 per cent, that weighed dailj- 1.56 per cent.Changes in the brain weighed dail}^: A moderate loss in weightoccurred during each of the first five da\'s, after which there was aslow, continuous loss up to the end of the month and beyond. Neitherthe first nor the second change of solution produced any efl'ect.SIXTY-FIVE PARTS 95 PER CENT ALCOHOL AND 35 PARTS 3 PER CENTFORMALIN.(Alcohol, 65; water, 34; formalin, 1.)End of first week: Weight of brains in 3 c. c. to the gram solutionhad fallen in average nearly 4 per cent. The lighter brain lost some-what more than the heavier one. The specimen in the 6 c. c. to thegram solution lost much more than either of the above; that weigheddaily lost slightly more than either of the other two in similar quantityof the preservative.End of first month: Weight of the two brains in 3 c. c. to the gramsolution had diminished but slightly, that of the specimen in 6 c. c.to the gram solution distinctly more, while that of the brain weigheddaily was equal.Changes in the brain weighed daily: The first day a slight (1.56 percent) rise, then a gradual loss; an insignificant rise the first and thirddays after the first change of solution, then stabilit}', with slight upsand downs. Mo rise or fall in weight after the one-month's change ofthe preservative. 298 PROCEEDINGS OF THE NATIONAL MUSEUM. NO. 1451. BRAI^^S AND BRAIN PRESERVATIVES?IIEDLICKA. 299 300 PROCEEDINGS OF THE NATIONAL MUSEUM. NO. 1451. BRAINS AND BRAIN PRESER VA TIYES?HRDLICKA. 301SODIUM ACETATE (FUSED), 130 GRAMS; SODIUM CHLORIDE, 110GRAMS; FORMALIN, 20 C. C; 95 PER CENT, ALCOHOL, 460 C. C;WATER, 540 C. C.(100 c. c. = sodium acetate, 13 ; sodium chloride, 11 ; alcohol, 46 ; water, 54 ; formalin, 2. ) End of first week: Weight of brains in the 3 c. c. to the gram solu-tion had diminished in average by 14 per cent, or one-seventh of theoriginal. Variation : From 81. 51 (original weight 103. 5 grams) to 86. 89per cent (original weight 103 grams) =2.35 per cent. The two heavi-est brains (118.2 and 115.2 grams) lost in weight, respectively, 13.28and 14.07 per cent, the two lightest ones (101 and 82 grams) 14.36 and14.64 per cent. The specimen in the 6 c. c. to the gram solution (origi-nal weight 110 grams) lost 15 per cent, that weighed every day (origi-nal weight 100.8 grams) 14.69 per cent, becoming each relativelylighter than any but one of the above.End of first month: The solution had been changed, both at the endof the first week and at the end of the first month, onl}" with the speci-men weighed daily, without, however, any material difference result-ing. Of the nine brains in the 3 c. c. to the gram solution eighthad, since the end of the first week, slightly increased in weight, whilein one the weight was the same. The gain ranges from 0.51 to 1.15per cent. Variation: From 85.51 to 87.38 per cent =1.87 per cent.The changes were quite alike. The specimen in the 6 c. c. to the gramsolution gained 0.45 per cent in weight, that weighed daily 2 per cent,or more than any other in the whole series.Changes in the brain weighed daily: A pronounced loss during thefirst twenty-four hours, the next da}^ a smaller loss, then three da3's ofstability, and then a slight loss again. After first change a slight riseduring the first twenty-four hours and lasting to next day, then aslight loss lasting four days and then slow rising. No marked effectof the second change of solution.SUMMARY.A glance at the foregoing data and at those of Part I of this papershows that, with the same preservatives, the results were in substancemuch alike, but that in the first series there was a much greater vari-ation in results.The simple formalin solutions all show, with ail brains, the sametype of effects, consisting of a sharp initial rise in the weight of thespecimens, reaching a maximum within less than a week, with a sub-sequent gradual, long-continued loss. The rise, very clearly shownby the tests on sheep brains, is related in an inverse ratio to thestrength of the formalin in the solution. The proportion of loss ismuch alike and is apparently independent of the formalin percentage,which makes it probable tliat it consists of simple solution by thewater of the preservatives. 302 PROCEEDINGS OF THE NATIONAL MUSEUM.The addition of common salt to formalin solutions acts veiy muchlike larger proportions of formalin alone. The initial rise is shortenedand reduced; in stronger concentrations it is replaced in twent3'-fourhours by loss; but the subsequent loss in weight proceeds much likethat in simple formalin solutions."Additions of alum to formalin solutions cause, though the specificgravities of the resulting liquids are less, a greater loss in brain weightthan the liquids with similar additions of common salt. The weakersolutions caused a smaller initial (one week) but a greater subsequentloss than the concentrated one.^The mixtures of alum, common salt, and formalin are characterizedby the great loss which they produce in the weight of the specimensafter the first week. There is no advantage whatever in these solu-tions.The three mixtures of alcohol with formalin all show an initial lossin the weight of the specimens, but subsequenth" there is a relativelygreat stability. Several of the groups (particularlj^ with Stroud'sliquid) show actually a little gain following the initial loss. As theproportion of the formalin in any of the solutions is quite insignificant(1 per cent), these effects must be referred nearly wholl}^ to the alco-hol and water in the solutions, with the action of which, so far as ourknowledge goes, thev agree. '^Individual variation was present with all the liquids used, most inthe 1.5 per cent formalin and the 1,080 specific gravity common saltsolution with 5 per cent formalin, least in the 'ob parts of alcohol with35 parts of 3 per cent formalin (two specimens only) and in the sodiumacetate-sodium-chloride-alcohol-formalin mixture. In a large major-it}' of the preservatives the variation was greater at the end of thefirst month than at the end of the first week; after that it still increasedwith some solutions, while with others it grew less.The most potent discernil)le cause of this individual variation was,as in the cases dealt with in the first part of this paper, the differencein size of the specimens. Another ascertainable cause, but operativeto a less extent, was the relative quantity of the preservatives. Evenwith the sheep brains alone the large ones suffered in the same rela-tive quantity of preservative less change, particularl}- less ultimateloss, than the small ones; and a double quantity of the liquid, eventhough most brains chosen for the experiment were large ones, resulted,in the majority of instances, in a loss of weight markedly greater thanthe average in the smaller proportion of the solution. The variationswhich remain unaccounted for are of obscure and probably complex ? A greater whiteness of the specimens was again noticeable.6 All the specimens showed very good hardening. The diniinution in size in thosein the concentrated ."Solution was very noticeable."See Donaldson, Jour. Morphol., 1894, p. 149. NO. 14.M. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 303nature; fortunately they are not, at least in normal animal brains,very serious.The effect of dail}' weighing the brain was, almost generally, greaterthan the average loss of weight.The changing of solution after one week and one month had in a fewinstances no appreciable effect, but mostly there was a consequent tem-porary (one to two days) rise in weight which acted as a retarder ofthe continuing loss.As to the practical results of these experiments on the value of variousbrain preservatives for macroscopical purposes, it is plain that neitheran}' of the simple formalin solutions nor any of those to which commonsalt or alum had been added, is satisfactory. The changes in theseliquids are considerable and their continuation prolonged, while thereare no compensatory advantages. No good purpose would be servedby using any of these mixtures, with one possible exception, in thefuture; the exception concerns the addition of alum to the solutionused for brains of foetuses or the very young, for the purposes ofincreasing the hardening.On the other hand, the results obtained with the alcohol and formalinmixtures are most encouraging. These liquids have produced butmoderate initial changes (much of which can be done awa}^ with byproper modifications of the solutions), followed by the all importantfeature of subsequent stability. The permanence of this stability hasnot received as yet a sufficient test of time, but Donaldson's prolongedobservations with other alcohol mixtures render it highly probable.The brain is not affected perceptibly by the necessary changes of solu-tion. At all events, it is with this class of preservatives that furtherexperiments are most justifiable.The addition of the salts in Stroud's liquid gives no superiority overthe simple alcohol-formalin solutions. The greater specific gravity ofthe mixture would commend it on account of the slightly greater pre-vention of deformation in the specimens, but the somewhat greaterinitial loss in weight and the subsequent continuous gain are disad-vantageous. If equally good results, so far as weight and size of thespecimens are concerned, can be obtained with simple mixtures, theseshould be preferred. Conservation of the form of a specimen in anypreservative is largel}^ a matter of proper care.As a result of the data obtained by the experiments reported uponin this paper, the tentative regulations below outlined concerning brainpreservation have been made in the laborator}- of ph3^sical anthropol-ogy of the U. S. National Museum. They can, it is hoped, be pur-sued with daily and longer periodical weighings of the specimens, andwith whatever modifications may become indicated in the liquids, untila substantiated and as simple as possible method of brain preservationhas been determined. It would be ver}' desirable if a concurrent 304 PROCEEDINGS OF THE NATIONAL MUSEUM.microscopical investio-ation could be made in some other laboratory,more suited for that purpose, as to the relative value of the variouspreservatives for the purposes of histology and pathology.PRESENT REGULATIONS CONCERNING BRAIN PRESERVATION IN THELABORATORY OF PHYSICAL ANTHROPOLOGY, UNITED STATESNATIONAL MUSEUM.Remove the brain as fresh as possible and with the least injury,without the dura; weigh at once; measure the proper solution into a jarof suitable size; place sufficient absorbent cotton on the bottom to pro-tect the specimen from deformation by pressure; in larger specimensintroduce a little cotton between the cerebrum and cerebellum, andlay the brain in, base downward. If necessary, support the hemi-spheres in proper position b}^ additional cotton; close jar as nearlyair-tight as possible and place on a shelf ovit of direct light of the sun;weigh, after the regular drainage, on the eighth day and change solu-tion. Weigh again on the thirt^^-first day, at the end of three monthsafter reception, and every three months (as long as the experimentslast). Solutionis and proportions. Formalin. Brains up to 50 grams in weight. . Brains 51 to 150 grams in weightBrains 151 to 300 grams in wi-ightBrains ;?)1 to ',t(X) grams in weightBrains above 900 grams in weight Distilled BRAINS AXD BRAIN PRESERVATIVES?HRDLICKA. 305ABSTRACT. Preservative. 3 per cent formalin5 per cent formalin10 per cent formalin!.'> per cent formalinSaturated .solution of common .salt with .5 per cent formalin1,030 sp. gr. common salt solution with 5per cent formalin1,015 sp. gr. common salt solution with 5per cent formalinSaturated solution of alum with 5 percent formalinOne-third saturated solution of alumwith 5 per cent formalinOne-fifth saturated solution of alum with5 per cent formalinOne-third .saturated solution of alumwith common .salt to 1,030 sp, gr. and 10per cent formalinOne-third saturated solution of alumwith common salt to 1,030 sp. gr. and 5per cent formalin80 parts of alcohol and 20 parts of 5 percent formalin6.5 parts of alcohol and 35 parts of 3 percent formalinSodium acetate, sodium chloride, forma-lin, and alcohol solution 3 c. c. per gram (weight at periods indicated below).Averageper centoforiginalweight atthe endof 1 week. 121. 4 117.9115.112. 8 92.997.9101.977.299. 4101.2 92.1 102.488.596.385.9 Percentvaria-tion. 5.43.94.08.94.910.3(1.2)7.53.54.0 3.66.2(1.1)2.3 Averageper centoforiginalweight atthe end of1 month121.3116.0113.1111.089. 3 95.9100.371.593.292.1 81.1 85.686.995.886.5 Percentvaria-tion. Averageper centoforiginalweight atthe end of2 months6.85.8 .5.39.86.29.9(1.4J10.47.17.9 8.1 .5.8(1.2)1.9 118.2112.8110.3106.686.89.3.69.5.971.088.085.2 71.2 76.387.096.086.4 Percentvaria-tion. 6.75.34.010.66.510.8(1.3)3.710.16.6 11.4 8.97.1(1.7)1.9 Preservative. 6 c. c. per gram (weight at j 3 c.periods indicated below) . Per cent of originalweight at the endof? c. per gram (weighedevery day). 1 week. 1 2month, months 35 Per cent of originalweight at the endof? 1 week. 1 2month, months 3 per cent formalin6 per cent formalin ,10 per cent formalin15 per cent formalinSaturated solution of common .salt with5 per cent formalin ,1,030 sp. gr. common salt solution with 5per cent formalin1,015 sp. gr. common salt solution with 5per cent formalinSaturated solution of alum with 5 percent formalinOne-third saturated solution of alumwith 5 per cent formalinOne-fifth saturated solution of alum with5 per cent formalinOne-third saturated solution of alumwith common salt to 1,030 sp. gr. and 10per cent formalinOne-third saturated solution of alumwith common salt to 1,030 sp. gr. and 5per cent formalin80 parts of alcohol and 20 parts of 5 percent formalin65 parts of alcohol and 35 parts of 3 percent formalinSodium acetate, sodium chloride, forma-lin, and alcohol solutionProc. N. M. vol. XXX?06 118.9118.9114.0109.193.294.3102.082.397.998.4 93.9 98.384.990.285.0 116.9116. 4 112. 8 102. 6 90.092.499.673.592.690.9 83.1 88.483.587.98.5.4 113.9111.4108.496.981.789.095.171.588.984.1 74.0 84.282.088.486.4 117.6117.8116.2108. 592.197.1101.374.295.8102.0 89.0 95.488.596.38.5.3 115.2116.0112. 2 104.7 94.298. 2570.385.92.3 78.6 86.687.196.387.3 113.2112.2110.399.6 91.293.469.379.281.3 73.3 81.083.09.5.3 20 C!i 306 PROCEEDINGS OF THE NATIONAL MUSEUM. VOL. XXX.CHANGES IN INDIVIDUAL SHEEP BRAINS.Preservative: 3 per cent formalin {3 c. c. per gram).[Condition of brain: Medium.] 70 per cent.Fig. -.'7 Proc. N. ? IN Ill'11-i-N III) I I r.i.M ii;i.?ir.vArivi:s CONTAINING roKMAi.iNSOLUTIONS CONTAINING ALCOHOL AND KOKMALIN MKAVIi;!; i.INKSProc. N. M. vol. XXX?OH. (To face page 306.) No. 27. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 307 Preservative: 5 per cent fortaalin {S c. c. per gram).[Condition of brain: Medium.] 308 PUOCKEDiyaS OF THE XATIOSAL MUSEUM. Preservative: 10 per cent formalm {3 c. c. per gram).[Condition of brain: Medium.] 2425..2627..2829..30..31..32..33" Date ofautopsy. Vjrain? Weight of Per cent [Weight of I Per centimmediatelyafter ex- I traction. brainafter 1week. 1905.June ' ....do. ....do. ....do.June I ....do'. ....do. ....do. ....do. ....do. Grams.110. 5 97.097.597.096.5112.0102.110.580.0125.0 Orams.127. 5 109.110.0112. 2111.5128.5117.5128.593.5142.6 oforiginalweight. 115. 38112.88112. 84115. 67115.54114. 73115. 19116. 29116.87114. 00 brainafter 1month. oforiginalweight. Grams.125. 5 107.0110.5108.109.5126. 5114.5127.92.5141.0 Per centof changebetweenthe end offirst andendof fourthweek. Additional weighings. 113. 57110. 31113.33111. 85113.47112.95112. 25114.93115. 63112. 80 Date. Weight!of brain -1.56 -2.28+ .45 -3.29 -1.79 -1.56 -2. 55 -1.17 -1.07 -1.05 1905.Aug. 7 ....do... ....do... ....do...Aug. 8 ....do.. ....do.. ....do.. ....do.. ....do.. Per centoforiginalweight. Gram.123.105.108.106.106.123.111.124.89,135, 111.31108. 25110.77109. 79109. 84109. 82109. 31112. 21111.25108. 40 aAt 6c. c. per gram.[Brain weighed daily. Received June 8, 1905. Original weight, 102 grams.] At the end ofday. Per centAb.solute { ofweight. 1 original I weight. FirstSecondTliirdFourtliFifthSixthSeventh 'IEighthN'inthTenthTwelfthThirteenth...FourteenthFifteenthSixteenthSeventeenthKiirhteentliNineteenthTwentiethTwenty-lirstTwentv-second .Twentv-third ...TwentV-fourth..Twentv-fifth....Twenty-sixth ...Twenty-seventhTwenty-eighth.. Grams.113.5116.0118.0118.0118. 5 119.0118.119.8120.120.0119. 2 119.0118.5117.117.5117.5117.5117.0116. 5 116.5116.011.5.511.5.0115.711.5.0115.0114.8 111.27113. 72115. 6S115.68116.17116.()7116. 17117. 45118.13117.64116.86116. 67116.17115. 19115.19115.19115. 19114.71114.21114. 21113.72113.23112.74113. 43112.74112.74112. 55 Change inpercentageof originalweightfrom dayto day. -4-11.27+ 2.45+ 1.96? .00+ .49 -t .00 - .49+ 1.28 At tlie end ofdav. Absoluteweight. + .68 .49 .78 .19 .50 .98 .00 .00 .00 .48- .50 .00 .49 .49 .49 .69 .69 .00 .19 Twentv-ninth ..Thirtieth"Thirty-firstThirty-second ..Thirtv-third...Thirtv-fourth..Thirty-fifthThirty-sixth...Thirtv-seventhThirtv-eighth .ThirtV-ninth ..FortiethForty-firstForty-second ..Forty-thirdFortv-fourth ..Forty-fifthForty-sixthForty-seventh.Forty-eighth ..Fortv-ninth .. . FiftiethFiftv-firstSixtiethSixty-seventh .Seventy-fourth Grams.114.7114.5116.116.0116.0115.5116.0115.6115. 5 115.0115.0114.5114.0114.0113.5113.113.0113.5113.0113.0113.0112.5113.0112.5III.O109. 5 ! Change inPer cent percentageof ; of originaloriginal I weightweight, from dayto day. 112. 47112.25113.72113. 72113.72113.23113.72113.23113. 23112. 74112. 74112. 25111.76111.76111.27111. 27110.78111.27110.78110. 78110. 78110.29110. 78110. 29108. 82107. 35 -o.os+ 1.47? .00? .00 - .49+- .49 - .49? .00 - .49? .00 - .49 - .49? .00 - .49? .00 - .49+ .49 - .49? .00i .00 - .49+ .49 - .49 -1.47 -1.47 a Change of solution. NO. 1451. BRAINS AND BRAIN PRESER VA TIVES?HRDLIVKA. 309 Preservative: 15 per cent formalin {3 c. c. per gram)[Condition of brain: Medium.] 310 PROCEEDINGS OF THE NA TIONAL MUSEUM. VOL. XXX. Preservative: Saturated mlution nf mlt, vntho per cent formalin {3 c. c. jier gram).[Condition of brain: Medium.] BRAim AND BRAIN PRESERVATIVES?HRDLIVKA. 311 Preservative: 1,030 sp. rp\ salt solution, iritJi 5 per cent formalin.[Condition of brain: Medium.] 312 PROCEEDINGS OF THE NATIONAL MUSEUM. Preservative: 1,015 sp. gr. salt wlufion nnth 5 per cent formalin {3 c. c. per gram).[Condition of brain: Medium.] NO. 1451. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 313 Preservative: Saturated solution of alum, witlt, 5 per cent formalin {S c. c. jyer gram)[Condition of brain: Medium.] 314 PllOCEEDINdS OF THE NATIONAL MUSEUM. VOL. XXX. Presmdliii': One-third saturated solution of alum, vithS per centformalin {3 c. c.jjenjram).[Condition of brain: Medium.] NO. KM. BRAINS AND BRAIN PRESERVATIVES?HRDLICKA. 315 Preservative: One-fifth safarated solution of alum, with 5 'per centformalin [Sec. j^er gram ) . [Condition of brain: Medium.] 316 PROCEEDINGS OF THE NATIONAL MUSEUM. VOL. XXX. Preservative: One-third saturated solution of alum, sodium chloride up to 1,030 sji, gr., with10 per cent formalin.[Conditiuii of brain: Medium.] NO. 1451. BRAINS AND BRAIN PRESER VA TI I'ES?HRDLIVKA . 317 Premrvallvc: Oite-tlnrd mlurated soiiUioti. of alum, vnth mlt uj> to 1,030 sp. gr.; 5 i^er centformalin [3 c. c. per gram).[Condition of brain: MQdium. 318 PROCEEDINGS OF THE NA TIONAL MUSEUM. Preservative: EigMij parts of 95 per cent alcohol and ?0 parts of 5 per cent formalin {3 c. c.per gram).[Condition of brain: Medium.] NO. 1451. BRAINS AND BRAIN PRESER VA TIVES?HRDLICKA. 319 Preservative: Sixty-five parts of 95 per cent alcohol and 35 parts of 3 per cent formalin.[Condition of brain: Medium.] 820 PROCEEDISGS OF THE XATIOXAL MUSEUM.Sodium acetate {fused) 130 grams; sodium chloride, 110 (/rams; formalin, 20 c. c; 95 percent alcohol, 4')0 c. c; water, 540 c. c {3 c. c. 2>er gram).[Condition of brain: Medium.] BRAIXS AXD BRA IX FRESERVATTVES?HRDLlrKA. MOuBIBLIOGRAPHY. V. Bardeleben, K., and U. Holzix (Oi'pekmaxn). Ein nenes Mittel zur ConsiTvi-rung von organis^chen Snbstancen, etc.: Verh. Ges. deutsch. Nat. und Aerzte, (: