S T U D I A G E O L O G I C A P O L O N I C AVol. 124, Kraków 2005, pp. 21–35.

Methods and Applications in MicropalaeontologyEd ited by J. Tyszka, M. Oliwkiewicz-Miklasiñska,

P. Gedl & M. A. Kaminski

Andrzej BOC ZA ROWSKI1

Chromo-stere os copy as a tool in mi cro pa lae on to logi calin ves ti ga tions: echi no derms as a case study

(Figs 1–4; Tab. 1)

Ab stract. An ana glyph tech nique, as a va ri ety of chromo-stere os copy, is pre sented and adapted forthe in ves ti ga tion pur pose. This method al lows per fect docu men ta tion of 3- dimensional ob jects,in clud ing all types of 3D mi cro/fos sils. Ana glyphs are es pe cially rec om mended for de scrib ing typecol lec tions and make them avail able world wide. The meth ods of stereo gram prepa ra tion by means ofa scan ning elec tron mi cro scope and ad ap ta tion of the tech nique to mi cro pa lae on tol ogy are de scribed.This use ful tech nique is tested on SEM im ages of com plex echi noid struc tures (or gans) calledpe di cel lar iae. The ob tained stereo grams docu ment their anat omy and help iden ti fi ca tion of the fos silma te rial as well as their com pari son to Re cent speci mens. Nu mer ous echi noid pe di cel lar iae fromBa jo cian and Batho nian de pos its of the ore- bearing clays of the Czês to chowa for ma tion have beenana lysed. Two sepa rate meth ods of ana glyph im age prepa ra tion are de scribed (for RGB and gray scale im age mode). The sam ples come from sev eral brick yards lo cated in the area of the Kraków- Czêsto-chowa Up land, Cen tral Po land. Pe di cel lar iae are rep re sented by glo bif er ous and ophicepha lous types. It has been proved that Ju ras sic echi noids al ready had very well de vel oped de fence strate gies, whichap plied pe di cel lar iae, quite of ten equipped with a venom- bearing sys tem simi lar to the one in Re centspeci mens. Large mor pho logi cal simi lar ity of the in ves ti gated Ju ras sic speci mens to the Re centrep re sen ta tives of the Pare chini dae fam ily, es pe cially to the ge nus Para cen tro tus Mortensen has been proved. How ever, no rep re sen ta tives of this type of echi noid have been found in Ju ras sic sedi ments sofar.

Key words: chromo-stereoscopy, Echi noi dea, anat omy, Mid dle Ju ras sic, Re cent, Po land, Croa tia.

IN TRO DUC TION

Or gans of hear ing and sight en able a hu man be ing to re ceive and ana lyse spa tialin for ma tion. Stereophony was de vel oped some time ago and be came an ob vi ous

1 Uni ver sity of Sile sia, Fac ulty of Earth Sci ences, De part ment of Pa lae on tol ogy & Bi os tra tigra phy, ul. Bêdzi ñska 60, 41- 200 Sos now iec, Po land. E- mail: boc zar@us.edu.pl

part of our life. Stere os copy is still a very lit tle known tech nique and ob tain ing aproper 3D im age still causes a lot of prob lems. The word ana glyph pre vi ouslymeant low- relief or na ments on gob lets, chal ices or pitch ers. The word origi natedfrom a Latin word ana gly phus, which means – made in low re lief or pressed, whichre lated to the light ef fect re sem bling 3D im ages. Cur rently the word is ap plied alsofor a stereo scopic il lus trat ing tech nique that uses the si mul ta ne ous pro jec tion oftwo stereo grams. Ana glyphs are one of the easi est meth ods of chromo stereo scopicre cord ing of 3D space on a 2D sur face. The main dif fer ence be tween chromo- stereoscopy and other tech niques is the way the im age is trans ferred through the eye to the brain; each im age from a given ste reo pair is fil tered by one of com ple men -tary col ours (McAl lis ter & Heb bar, 1991). The im ages as signed for the left and theright eyes are su per im posed and var ied in col our. They are ob served throughglasses with spe cial fil ters, where the left glass is most fre quently red and the rightone is blue, cyan or green.

The first in ter ests in the stere os copy tech nique date back to the XVI cen tury.The real de vel op ment of the tech nique ap peared in the XIX cen tury to gether withthe be gin nings of pho tog ra phy. Roll mann con sti tuted the ba sis of stere os copy in1853 (www.stere os copy.com). The first chromo stereo scope was de signed byD’Al meida in 1858. It was adapted for prac ti cal us age by du Hauron in France in1891 (www.stere os copy.com). Stere os copy was a popu lar en ter tain ment in townsin the thir ties of the last cen tury. Later it was al most com pletely for got ten. Cur -rently due to de vel op ment of digi tal tech niques it may be come popu lar again.

Our brain re ceives op tic im pulses, analy ses them and pro cesses into a 3D im age. Three- dimensional vi sion was fixed by evo lu tion in our an ces tors in the or der Pri -mates at the turn of late Pa laeo cene and early Eo cene, to gether with ap pear ance offirst pro simi ans. The fea ture oc curred much ear lier in some preda tors. Stereo scopic vi sion of the sur round ing world is used by or gan isms to de ter mine per fectly the dis -tance be tween ob jects and per sons ob serv ing them (Car roll, 1988). Such evo lu -tional ad ap ta tion of the body of rep re sen ta tives of the pri mates was forced by liv ingamong tree branches. When mov ing through the crowns of trees, proper es ti ma tionof dis tance played a cru cial role in their chance to sur vive. It re duced a field of vi -sion, but this short age was com pen sated with ex treme mo bil ity of the neck. Im agesre ceived by a left and right eye is ac quired from two dif fer ent points of view, whichdif fer from each other with par al laxes (Ostafic zuk, 1978). A 3D im age is a true pic -ture of re al ity. Geo met ric fea tures, such as size, shape, sym me try, lo ca tion and il lu -mi na tion play sig nifi cant roles among vari ous fea tures of ob jects, which we ac -quire. Ap pli ca tion of 3D il lus tra tions in natu ral sci ence seems to be un lim ited, but it is very rarely used. Stereo pairs in mi cro pa lae on tol ogy is only ap plied rarely, mostno ta bly in the Stereo- Atlas of Os tra coda (Boomer et al., 1973–1998), as well as inde scrip tion of type speci mens of holo thu ri ans (Boc za rowski, 1997). In the days ofim me di ate re sults ob tained by digi tal pho tog ra phy, pre par ing an ana glyph it self isex tremely easy, more dif fi cult is ob tain ing a proper ste reo pair of pho to graphs. Therec og nis ing abil ity of a healthy hu man eye en ables to dis tin guish two points be ingin a dis tance of 20 µm.

22 A. BOCZAROWSKI

THE ANA GLYPH METHOD

When ap ply ing stere os copy for small ob jects, an im age can be dis tin guishedwith the same re solv ing power, but this is not suf fi cient. It is pos si ble to make ste reo pairs ap ply ing mi cro scope tech niques. In the case of a scan ning elec tron mi cro -scope (SEM), the left im age is the ba sic one, the sec ond im age is turned left by thean gle of 6°–12°. The im age may be also turned right but in that case be fore mak ingan ana glyph the places of im ages should be ex changed. The smaller an gle is usedwhen the ob ject is very sa li ent and strongly var ied mor pho logi cally. A larger an gleis ap plied when pho to graphed speci mens are al most flat. The author used an en vi -ron mental scan ning elec tron mi cro scope Phil ips XL30 ESEM. This equip ment al -lows tak ing pic tures of large pa lae on to logi cal ob jects en larged even four times andin a large field of vi sion. Hence, a con cept of SEM mac ro pho tog ra phy may be in tro -duced to pho tog ra phy. Field pic tures are taken from two places at dif fer ent an glesin re la tion to the cen tre of the im age. In the case of re mote pic tures the dis tance issev eral me ters, in the case of close pic tures it is about a few cen ti me tres. Graphicpro grams, such as Adobe Pho toshop® 7 or 8, CorelPHOTO- PAINT® 9, 10, 11 or12, Jasc Paint Shop™ Pro® 9 or Mi cro grafx® Pic ture Pub lisher 10, trans fer pre -pared pic tures, where proper col ours are ap plied (red and cyan). A graphic pro gramwith RGB chan nel sepa ra tion or a spe cial pro gram for ana glyphs may be ap pliedfor the pur pose. The fur ther step is to pro cess im ages into semi- transparent ones and to draw them aside un til the stereo scopic ef fect is ob tained (Dubois, 2001).

The im pres sion of per spec tive may be ar ti fi cially en hanced by far ther par al lelsepa ra tion of the im ages (Sand ers & McAl lis ter, 2003). De pend ing on the im agereso lu tion, the one proc essed by the red fil ter should be moved left by a propernumber of pix els un til the ef fect of sink ing the im age be low the pro jec tion sur facebe ob tained. The pro jec tion sur face means a piece of pa per, a moni tor screen or ascreen where im ages are pro jected. Su per im pos ing of both im ages may cause theim pres sions that the pho to graphed ob ject is ly ing on the pro jec tion sur face or seems to be be low it. Mov ing the red im age to the right till sepa ra tion of a ste reo pairmakes the im pres sion the ob ject is above the pro jec tion sur face. Change of im ageplaces causes in ver sion of the ap par ent re lief of the ob served ob ject. Ap proxi mateeye spac ing of a hu man be ing is equal to 65 mm. An gle reso lu tion of a hu man eye atstereo scopic vi sion is 10’’. This means, that per cep ti ble dif fer ence in dis tance in -creases with the range of stereo scopic vi sion (Ostafic zuk, 1978). This fact is thecause of a sig nifi cant limi ta tion in ob ser va tion of larger stereo scopic im ages bymeans of op ti cal in stru ments. The range of stereo scopic vi sion may be ar ti fi ciallyen larged by elon ga tion of the eye base by ap pli ca tion of spe cial op ti cal equip ment,but such a method is very ex pen sive. The ana glyph method is cheap and has no such limi ta tions and may be used to ob serve very big im ages from vari ous dis tances.Sepa ra tion of par ticu lar il lus tra tions of each ste reo pair leads to the ef fect whentheir mar ginal parts are not well visi ble and the stereo scopic ef fect van ishes. Suchil lus tra tions should be prop erly cut. In case of pa lae on to logi cal ap pli ca tions, it isrec om mended to sepa rate the pho to graphed ob jects from the ma trix and to place

CHROMO-STEREOSCOPY 23

them on an ideal black back ground. This will help to avoid the nega tive ef fect ofvan ish ing of the stereo scopic ef fect in mar ginal parts of the im age. A se quence ofmany im ages show ing revo lu tion of the fig ure or pano ramic im ages al lows mak ingfilm clips or vir tual ani ma tions. Digi tally pre pared stereo grams may be eas ily ap -plied for vari ous pres en ta tion tech niques. Col our cali bra tions of the pro ject ingequip ment and black out in the pro jec tion room are nec es sary when watch ing thestereo grams on a large screen.

AP PLI CA TION OF ANA GLYPH METHOD

In ves ti ga tion of echi noid pe di cel lar iae

Ap pli ca tion of ana glyphs to docu ment echi no derms en ables quick pres en ta tionof their very com pli cated mor phol ogy. The de scribed method has been ap pliedeven with bet ter ef fects to carry out mi cro scopic in ves ti ga tions of sin gle echi no -derm scler ites. Skele tal ele ments of the echi no derm scleri tome are usu ally built ofmany walls and their shapes are very com pli cated. Pre pared ana glyphs en able spa -tial in ves ti ga tion of the speci men’s mor phol ogy with out any so phis ti cated tech ni -cal equip ment. Quick com pari son of sev eral im ages of the same or dif fer ent speci -mens is also pos si ble (Figs 1–4) to pres ent as many pic tures as may be ana lysed si -mul ta ne ously. This is very dif fi cult or even im pos si ble when other meth ods are ap -plied.

In this work, chromo- stereoscopic tech niques have been tested and ap plied toin ves ti gate the pe di cel lar iae of Echi noi dea. The pe di cel lar iae are small, cal care ous, highly spe cial ised, pin cer like or gans cov er ing echi noid tests and as ter oid’s bod ies.Those mi cro scopic scler ites may con tain venom tis sues. Echi noids use pe di cel lar -iae to de fend them selves against preda tor at tempts to set tle on their tests and toclean their test. Four main types of echi noid pe di cel lar iae may be dis tin guished(Tab. 1). It must be con sid ered that an in di vid ual may have all ba sic types of pe di -cel lar iae on its test (Boc za rowski, 2001, 2004a, b).

Ma te rial

Hun dreds of kgs of rock ma te rial from four sec tions of Ba jo cian through Batho -nian de pos its of the Kraków- Czêstochowa Up land were col lected for Echi noids.Ad di tional re cent ma te rial from Croa tia (Krk Is land) was in ves ti gated.

The Echi noid speci mens have been placed in the sci en tific col lec tion of the De -part ment of Pa lae on tol ogy and Bi os tra tigra phy, Fac ulty of Earth Sci ence, Uni ver -sity of Sile sia un der the fol low ing num bers: GIUS 8–1554, 1557, 2471 Gna. –brick yard Gnaszyn (Czês to chowa); GIUS 8–1825, 1830 Sow. – so called “Sowa”brick yard (Czês to chowa); GIUS 8–2074 Bla. – brick yard Bla nowice (Zawier cie);GIUS 8–2269, 2270 Wrz. – brick yard Wrzosowa; GIUS 12–2511 Krk. – Krk Is -land, Croa tia. Their lo ca tion and bi os tra tigraphic cor re la tion can be found in Ma -tyja and Wierzbowski (2003) Anat omic ter mi nol ogy is pre sented in Figs 1–4.

24 A. BOCZAROWSKI

Re sults

Nu mer ous, well- preserved echi noid pe di cel lar iae have been found in mac er ated Mid dle Ju ras sic ore- bearing clays of the Czês to chowa For ma tion. Their oc cur rence is very high in Bla nowice and Gnaszyn sec tions (Boc za rowski, 2004a, b; Boc za -rowski in Gedl et al., 2003, 2004). Glo bif er ous and ophicepha lous pe di cel lar iae,

CHROMO-STEREOSCOPY 25

Ta ble 1

Com pari son of ba sic fea tures of li ving and fos sil echi noid pe di cel lar iae(af ter Boc za rowski, 2001, 2003, 2004a, modi fied)

Features

Type of pedicellariae

GlobiferousFigs 3: 1–3, 4: 2,

10

OphicephalousFigs. 1, 2, 4: 1, 3,

4, 11Triphyllous

TridentateFigs 3: 4–6, 4: 9

Blade teeth general many sharp many blunt absentmany sharp and

blunt

Terminal teethone or more,

venomabsent absent single

Subterminal teeth present absent absent absent

Central grasping tooth in the centre ofgripping area

absent present absent absent

Distal point of thevalve contact

present: punctualcontact

present: pick oredge contact

absentpresent: pick or

edge contact

Valve edgedenticulation

various present absent various

Handle absent present absent absent

Typical number of the head valves

3 3 2 or 3 3

Number of the headvalves (min.-max.)

2–5 2–3 2–4 2–5

Venom tissue present various absent absent

Angle between closed valves

120° or other 120° or other180°, 120° or

other120° or other

Opening angle 90°–180° 180° 180° 180°

Autotomy present ? ? ?

Activity stimulating stimulating continuous stimulating

Stimulation chemical mechanical spontaneous mechanical

Functionsdefensive – using

venom againstbiological stimuli

defensive ,against

mechanicalstimuli

protective – keepthe test clean,

rarely defensive

defensive –against biological

stimuli

be long ing to sev eral mor pho types have been found there. Glo bif er ous pe di cel lar -iae usu ally con sist of three per fo rated valves (Fig. 3: 1–3). The base pro trudes in the form of a proxi mal base knob. A nar row keel sepa rates the valves. Keel ridges turninto a blade to wards dis tal and ad ax ial di rec tions. Lat eral and cen tral cog- teeth ap -pear close to the valve base (Fig. 3: 1–3). Tri valve ophicepha lous pe di cel lar iaehave typi cal spoon- like blade with minute den ticu la tion on the dis tal edge (Figs 1,2). The short keel di vides two el lip tic hol lows where ad duc tor mus cles were at -tached. Lat eral and cen tral cog- teeth oc cur in the wid est part of the valve. To wardsthe proxi mal edge they con nect to gether form ing ho moge nous trans verse ridges.The valve han dles may be sym met ric or not and may have a han dle fo ra men.

In ter pre ta tion and dis cus sion

Echi noids from Mid dle Ju ras sic de pos its in Po land have been rarely de scribed(Laube, 1867; Jesionek- Szymañska, 1963, 1980), but their oc cur rence has beenmore fre quently men tioned (Reh binder, 1913; Ró¿ycki, 1953). Blake (1968) de -scribed the old est pe di cel lar iae, which be long to two Si lu rian echi noids (Lud low,Here ford shire, Great Brit ain). Mortensen (1928–1951), in his mono graph, de ter -mined pe di cel lar iae as a ba sic ele ment in taxo nomic in ves ti ga tions (see also Jensen, 1982). De tailed in for ma tion on the func tions and re search his tory of the pe di cel lar -iae were car ried out by Geis (1936), Camp bell (1973, 1974, 1976, 1983), Law rence(1987) and Boc za rowski (2001, 2003, 2004a, b).

Com plete tests are rarely found in Pol ish Mid dle Ju ras sic, while dis ar ticu latedre mains are quite com mon. The struc ture of pe di cel lar iae is simi lar to the one ofspines as they evo lu tion ary de rive from them (Blake, 1968; Boc za rowski, 2001,2003, 2004a, b). Suc ces sive strat egy to frighten ene mies with venom has been ap -plied by echi noids at least since the De vo nian (Boc za rowski, 2001). The fre quentoc cur rence of those re mains in fos sil sedi ments are due to the fact that pe di cel lar iaeper formed mass au totomy in case of dan ger. Ju ras sic and Cai no zoic forms had wide blades with a deep blade sul cus, but de creased sur face of a venom gland. Ad di tion -ally, the venom glands could oc cur on the stems. The wide stems al lowed for thequick in jec tion of poi son into mi cro scopic dam ages made by ter mi nal and sub ter -mi nal venom teeth.

Com pari son of the dis cussed pe di cel lar iae from the Mid dle Ju ras sic of the studyarea with Re cent forms proves simi lari ties be tween glo bif er ous (mor pho type I) and ophicepha lous pe di cel lar iae (mor pho type III) and rep re sen ta tives of the Pare chini -

26 A. BOCZAROWSKI

Fig. 1. Mor phol ogy of the ophicepha lous pe di cel lar iae of Para cen tro tus lividus (La marck) fromKrk Is land (Re cent, Croa tia). 1 – Two valves in ana tomi cal po si tion, ad ax ial view, GIUS 12–2511Krk. P01- E- 004; 2 – Spe cial pre pared two valves in ana tomi cal po si tion, in ner view, GIUS 12–2511Krk. P01- E- 044. 3 – Sin gle valve, ad ax ial view, GIUS 12–2511 Krk. P01- E- 007. 3D ana glyphim ages: 1a, 2a, 3a (the stereo scopic ef fect will be visi ble with red-blue glasses only); 2D grey scaleim ages: 1b, 2b, 3b. Pho to graphs taken by author with Phil ips XL30 ESEM

CHROMO-STEREOSCOPY 27

dae fam ily, e.g. Para cen tro tus gai mardi (Blain ville), re cently known from theSouth Af ri can At lan tic coast (Mortensen, 1928–1951; Cher bon nier, 1959) orPara cen tro tus lividus (La marck) from the Medi ter ra nean Sea (Cher bon nier, 1968). At pres ent, only rep re sen ta tives of the Pare chini dae Fam ily have a blade with ablade sul cus. The prob lem is that the old est rep re sen ta tives of the fam ily are notknown be fore the Mio cene – Isech inus Lam bert; Plio cene – Psam mech inus Agas -siz and De sor; or Pleis to cene – Para cen tro tus Mortensen, Parechinus Mortensen(af ter Smith, 2004). The de scribed mor pho types of Ju ras sic pe di cel lar iae areknown re cently e.g., at Ar ba cii dae (Ar ba cia Gray), which also oc curred in Mid dleJu ras sic. Rad wañska (1999) de scribed rep re sen ta tives of the or der i.e. Ac ro pel tis L. Agas siz and Glyp ti cus L. Agas siz but not be fore the early Kim me ridgian in Po land.They have not yet been dis tin guished in the Mid dle Ju ras sic. The mor pho type II re -sem bles Ju ras sic pe di cel lar iae Pe lan ech inus Keep ing (Smith 2004). A speci men ofPe lan ech inus co ral linus (Wright) from the Ox for dian in York shire has com pleteophicepha lous and triteeth pe di cel lar iae, how ever the lat ter ones have not beenfound in the Ju ras sic of Po land. Smith (2004) in cludes here Pe lan ech inus oo li ti cum(Hess) from the Swiss Ba jo cian (=Pe la no dia dema Hess, 1971, 1972, 1973, 1975).Tiny parts of test ac com pa ny ing the pe di cel lar iae do not en able to de ter mine theirproper ori gin. The ap pli ca tion of the chromo stereo scopic method helped to in ves ti -gate anat omy of Ju ras sic pe di cel lar iae and their sys te mat ics, as well as their com -pari son to Re cent speci mens.

CON CLU SIONS

Chromo- stereoscopy can be ap plied dur ing prepa ra tion of pa lae on to logi caldocu men ta tion. Method de scribed here can be used with suc cess in print ing, pres -en ta tions, slide or nega tive films, mov ies and Inter net sites. Ana glyphs are verysuit able to il lus trat ing holo types and type ma te rial par ticu larly. This is really a verysim ple, quick and in ex pen sive way to re ceive three- dimensional im ages. All ana -glyph im ages should be viewed with spe cial red- blue glasses. The ste reo pair can be made by SEM, ESEM, photo cam era, pair of cam eras or by 3D soft ware (e.g., 3DStu dio Max™, Me taCrea tions™ Bryce, Maya® or Vue d’Esprit™).

28 A. BOCZAROWSKI

Fig. 2. Mor phol ogy of the ophicepha lous pe di cel lar iae of Para cen tro tus lividus (La marck) fromKrk Is land (Re cent, Croa tia). 1 – Whole head, abax ial view, GIUS 12–2511 Krk. P01- E- 042. 2 –Whole head, op po site side of an other speci men, abax ial view, GIUS 12–2511 Krk. P01- E- 041. 3 –Sin gle valve, abax ial view, GIUS 12–2511 Krk. P01- E- 023. 4 – Head base, outer proxi mal view,GIUS 12–2511 Krk. P01- E- 026. 3D ana glyph im ages: 1a, 2a, 3a, 4a (the stereo scopic ef fect will bevisi ble with red-blue glasses only); 2D gray scale im ages: 1b, 2b, 3b, 4b. Pho to graphs taken by authorwith Phil ips XL30 ESEM

CHROMO-STEREOSCOPY 29

Pro ce dure of mak ing the ana glyph im ages were made with Adobe Pho toshop® 7.0(Win dows plat form)

Method for sepa rate ob jects of gray scale mode im ages (©Andrzej Boczarowski):01) make the im age ste reo pair;02) open the left and right im ages;03) ac ti vate “lay ers win dow”: WIN DOW LAY ERS;04) sepa rate ob jects from back ground, do the same with both im ages;05) save these im ages as *.psd files: FILE SAFE AS (= Ctrl + Shift + S);06) go to the first im age (left im age) and click the layer with sepa rate ob ject;07) than: SE LECT ALL (= Ctrl + A);08) and next: EDIT COPY (= Ctrl + C);09) cre ate a new file: FILE NEW (= Ctrl + N);10) in “dia log menu” choose name of new file, be sure that the new im age will have cor rect

reso lu tion and RGB col our mode;11) paste cop ied file as a new layer: EDIT PASTE (= Ctrl + V);12) la bel this layer as “left im age”: LAYER LAYER PROPERI TIES;13) se lect the sec ond im age, copy and paste it as a next layer (up per most);14) la bel this layer as “right im age”: LAYER LAYER PROPERI TIES;15) go to the top layer and change the opac ity to 50%, the top im age will be slightly trans par ent,16) align pre cisely fo cus area of both im ages, by mov ing and ro tat ing top layer;17) re store opac ity of top layer to 100 %;18) se lect “right im age” layer;19) and next: IM AGE AD JUST MENTS LEV ELS (= Ctrl + L);20) se lect the RED chan nel: IN PUT LEV ELS RED (= Ctrl + 1)21) go to the OUT PUT LEV ELS and change the 255 to 0 (= Tab, Tab, Tab, Tab, “0”);22) click OK (= En ter), the col our of this layer should be cyan;23) se lect “left im age” layer;24) and next: IM AGE AD JUST MENTS LEV ELS (= Ctrl + L);25) se lect the GREEN chan nel: IN PUT LEV ELS GREEN (= Ctrl + 2)26) go to the OUT PUT LEV ELS and change the 255 to 0 (= Tab, Tab, Tab, Tab, “0”);27) do the same with BLUE chan nel: IN PUT LEV ELS BLUE (= Ctrl + 3); (= Tab, Tab, Tab,

Tab, “0”);28) click OK (= En ter), the col our of this layer should be red;29) se lect top layer again;30) open the top menu “set the blend ing mode for the layer” or: LAYER LAYER STYLE

BLEND ING OP TIONS and change op tion GEN ERAL BLEND ING from “nor mal” to“screen”;

31) re size im age can vas to proper size: IM AGE CAN VAS SIZE;32) choose “back ground layer” and fill it as black;

30 A. BOCZAROWSKI

Fig. 3. Mor phol ogy of the glo bif er ous and tri den tate pe di cel lar iae of Para cen tro tus lividus(La marck) from Krk Is land (Re cent, Croa tia). 1–3 – Sin gle valves of glo bif er ous pe di cel lar iae: 1 –ad ax ial view, GIUS 12–2511 Krk. P01- E- 027. 2 – Abax ial view, GIUS 12–2511 Krk. P01- E- 029. 3 –Lat eral view, GIUS 12–2511 Krk. P01- E- 028. 4–6 – Sin gle valves of tri den tate pe di cel lar iae: 4 –ad ax ial view, GIUS 12–2511 Krk. P01- E- 030. 5 – Lat eral view, GIUS 12–2511 Krk. P01- E- 031. 6 –Abax ial view, GIUS 12–2511 Krk. P01- E- 033. 7 – Stem of some pe di cel laria, side view, GIUS12–2511 Krk. P01- E- 011. 3D ana glyph im ages: 1a, 2a, 3a, 4a, 5a, 6a, 7a (the stereo scopic ef fect willbe visi ble with red-blue glasses only); 2D gray scale im ages: 1b, 2b, 3b, 4b, 5b, 6b, 7b. Pho to graphstaken by author with Phil ips XL30 ESEM

CHROMO-STEREOSCOPY 31

33) save im age as *.psd, *.tif, *.jpg or other for mat: FILE SAFE AS (= Ctrl + Shift + S);34) con trol the new ana glyph im age us ing red – blue glasses;35) ready.

Method for RGB mode im ages (© Andrzej Boc za rowski):01) make the im age ste reo pair;02) open the left and right im ages;03) be sure that the both im ages are in RGB col our mode: IM AGE MODE RGB COLOR;04) ac ti vate “chan nels win dow”: WIN DOW CHAN NELS;05) choose “left im age” and click red chan nel edi tion;06) than: SE LECT ALL (= Ctrl + A);07) and next: EDIT COPY (= Ctrl + C);08) choose “right im age” and click the red chan nel edi tion;09) re place the red chan nel of right im age by the red chan nel of left im age: EDIT PASTE (=

Ctrl + V);10) mark visi bil ity of all chan nels (= Ctrl + ~) but still hold the red chan nel edi tion;11) and next: SE LECT ALL (= Ctrl + A);12) align pre cisely fo cus area of both im ages, by mov ing and ro tat ing red chan nel;13) crop new im age to proper size;14) save im age as *.psd, *.tif, *.jpg or other for mat: FILE SAFE AS (= Ctrl + Shift + S);15) con trol the new ana glyph im ages us ing red – blue glasses;16) ready.

Ad di tional in for ma tion is avail able on the www sites:http://dog feath ers.com/3d/3dhowto.html;http://www.scec.org/ge ow all/makeana glyph.htmlhttp://nexel.stu dent.ut wente.nl/ran dysi mons.com/soft ware/3d/ – Cre ate your own 3D im agehttp://www.fear god.net/3dhowto.php – Gra ham’s Guide to 3D Pho tog ra phyhttp://www.mind help.com/_3d/how2make.htm – Cre at ing & Dis play ing Ste reo 3D

32 A. BOCZAROWSKI

Fig. 4. Echinoid skeletal elements. 1–4 – Pedicellariae from the Bathonian of the Kraków-Czêstochowa Upland, brickyard Gnaszyn (Czêstochowa), Ore-bearing Czêstochowa clay formation,Central Poland. 1 – Ophicephalous pedicellaria, morphotype III (after Boczarowski 2004a), adaxialview, GIUS 8–2471 Gna.\P13\E–401. 2 – Globiferous, morphotype I (after Boczarowski 2004a),adaxial view, specimen with broken blade, GIUS 8–2471 Gna.\P13\E–420. 3 – Ophicephalouspedicellaria, morphotype II (after Boczarowski 2004a), adaxial view, GIUS 8–2471Gna.\P13\E–410. 4 – Ophicephalous pedicellaria, morphotype I (after Boczarowski 2004a), adaxialview, GIUS 8–2471 Gna.\P13\E–414. 5-15 – Echinoid skeletal elements and their distribution on thetest of Paracentrotus lividus (Lamarck) from Krk Island (Recent, Croatia). 5 – Test, side view, GIUS12–2511 Krk. P01-E-052. 6 – Medium size secondary spine from the ambulacrum, side view, GIUS12–2511 Krk. P01-E-048. 7 – Small secondary spine from the ambulacrum, side view, GIUS 12–2511 Krk. P01-E-049. 8 – Rosette from sucking disc of tube foot, proximal view, GIUS 12–2511 Krk.P01-E-035. 9 – Single valve of tridentate pedicellaria, abaxial view, GIUS 12–2511 Krk. P01-E-033.10 – Single valve of globiferous pedicellaria, abaxial view, GIUS 12–2511 Krk. P01-E-029. 11 –Single valve of ophicephalous pedicellaria, abaxial view, GIUS 12–2511 Krk. P01-E-023. 12 –Secondary minute spine, side view, GIUS 12–2511 Krk. P01-E-050. 13 – Stem of some pedicellaria,side view, GIUS 12–2511 Krk. P01-E-011. 14 – Big secondary or regenerated primary spine from theinterambulacrum, side view, GIUS 12–2511 Krk. P01-E-003. 15 – Primary spine from the inter-ambulacrum, side view, GIUS 12–2511 Krk. P01-E-001. 3D anaglyph images: 1a, 2a, 3a, 4a, 5 (thestereoscopic effect will be visible with red-blue glasses only); 2D grayscale images: 1b, 2b, 3b, 4b,6-15. Photographs taken by author with Philips XL30 ESEM

CHROMO-STEREOSCOPY 33

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3D.http://nexel.stu dent.ut wente.nl/ran dysi mons.com/soft ware/3d/ – down loaded in 2004. Cre ate your

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