natgillin/translations-raw huggingface dataset:
Chemical exposure duration
F – izrasline s trbušne strane u odnosu na E. Obično malene i u parovima.
Biological endpoints
Prisutne u mužjaka iz kontrolnih skupina i ženki izloženih androgenima.
survival
Obrazac za ocjenu izraslina:
behaviour
Brojčana ocjena
fecundity
Datum
sex characteristics
1 – prisutne
optionally gonadal histopathology
Ukupan broj bodova
Test acceptability
2 – uvećane
Dissolved oxygen ≥ 60 % of saturation; mean temperature of 25 ± 2 °C; 90 % survival of fish in the controls; measured test concentrations within 20 % of mean measured values per treatment level.
3 – izražene
Dissolved oxygen ≥ 60 % of saturation; mean temperature of 26 ± 2 °C; 90 % survival of fish in the controls; measured test concentrations within 20 % of mean measured values per treatment level.
Dodatak 5.B
SOME CHEMICAL CHARACTERISTICS OF ACCEPTABLE DILUTION WATER
PROCJENA SEKUNDARNIH SPOLNIH OBILJEŽJA U MEDAKE RADI OTKRIVANJA ODREĐENIH ENDOKRINO AKTIVNIH KEMIKALIJA
COMPONENT
U nastavku se opisuje mjerenje papilarnih procesa [1] koji su sekundarno spolno obilježje u medake (Oryzias latipes).
CONCENTRATIONS
Nakon vađenja jetre (Dodatak 6.) truplo se stavlja u epruvetu s približno 10 ml 10-postotnog neutralnog puferiranog formalina (glavom prema gore i repom prema dolje).
< 20 mg/l
Ako je gonada fiksirana u otopini koja nije 10-postotni neutralni puferirani formalin, s pomoću skalpela napravite poprečni rez preko trupla između prednjeg dijela podrepne peraje i anusa, pazeći pritom da ne oštetite gonoporu i samu gonadu (slika 3.).
< 2 mg/l
Stavite glavu s prednjim dijelom ribe u otopinu za fiksiranje radi očuvanja gonade, a repni dio u 10-postotni neutralni puferirani formalin prema prethodnom opisu.
Unionised ammonia
Nakon što ste ribu stavili u 10-postotni neutralni puferirani formalin, pincetom uhvatite prednji dio podrepne peraje i držite ga presavijenog oko 30 sekundi kako biste raširili podrepnu peraju.
< 1 μg/l
Pri hvatanju podrepne peraje pincetom uhvatite i nekoliko šipčica peraje u prednjem dijelu, pazeći pritom da ne ogrebete papilarne procese.
< 10 μg/l
Nakon što je podrepna peraja bila raširena 30 sekundi, pohranite tijelo ribe u 10-postotni neutralni puferirani formalin na sobnoj temperaturi do mjerenja papilarnih procesa (mjerenje je potrebno provesti nakon najmanje 24-satnog fiksiranja).
Total organophosphorus pesticides
Mjerenje
< 50 ng/l
Nakon fiksiranja tijela ribe u 10-postotnom neutralnom puferiranom formalinu u trajanju od najmanje 24 sata, izvadite truplo ribe iz epruvete i obrišite formalin filtarskim papirom (ili papirnatim ručnikom).
< 25 ng/l
Položite ribu trbuhom prema gore.
Appendix 4A
Škaricama pažljivo zarežite podrepnu peraju (poželjno je, osim podrepne peraje, odrezati i mali dio perajne potpore).
SPAWNING SUBSTRATE FOR ZEBRAFISH
Pincetom uhvatite prednji dio odrezane podrepne peraje i stavite ga na stakalce s nekoliko kapi vode.
Spawning tray
Potom podrepnu peraju pokrijte pokrivnim stakalcem.
all glass instrument dish, for example 22 × 15 × 5,5 cm (l × w × d), covered with a removable stainless steel wire lattice (mesh width 2mm).
Pazite da pri hvatanju podrepne peraje pincetom ne ogrebete papilarne procese.
The lattice should cover the opening of the instrument dish at a level below the brim.
Prebrojite kosti bodlje s papilarnim procesima s pomoću brojača pod biološkim mikroskopom (uspravni ili invertni mikroskop).
On the lattice, spawning substrate should be fixed.
Papilarni procesi uočavaju se kad je mala nakupina procesa vidljiva na stražnjem dijelu kosti bodlje.
It should provide structure for the fish to move into.
U obrazac zabilježite broj kosti bodlje s papilarnim procesima na svakoj šipčici peraje (primjerice, prva šipčica peraje: 0, druga šipčica peraje: 10, treća šipčica peraje: 12 itd.) te u tablicu u Excelu unesite zbroj tih brojeva prema pojedinačnim ribama.
For example, artificial aquaria plants made of green plastic material are suitable (NB: possible adsorption of the test chemical to the plastic material should be considered).
Po potrebi fotografirajte podrepnu peraju i na fotografiji prebrojite kosti bodlje s papilarnim procesima.
The plastic material should be leached out in sufficient volume of warm water for sufficient time to ensure that no chemicals may be disposed to the test water.
Nakon mjerenja stavite podrepnu peraju u epruvetu kako je opisano u točki 1. i pohranite je.
When using glass materials it should be ensured that the fish are neither injured nor cramped during their vigorous actions.
Slika 1.
The distance between the tray and the glass panes should be at least 3 cm to ensure that the spawning is not performed outside the tray.
Dijagram s prikazom spolnih razlika u obliku i veličini podrepne peraje.
The eggs spawned onto the tray fall through the lattice and can be sampled 45-60 min after the start of illumination.
A – mužjak; B – ženka.
The transparent eggs are non-adhesive and can easily be counted by using transversal light.
Slika 2.A
When using five females per vessel, egg numbers up to 20 at a day can be regarded as low, up to 100 as medium and more than 100 as high numbers.
procesi na kostima bodlje šipčice podrepne peraje.
The spawning tray should be removed, the eggs collected and the spawning tray re-introduced in the test vessel, either as late as possible in the evening or very early in the morning.
J.P.
The time until re-introduction should not exceed one hour since otherwise the cue of the spawning substrate may induce individual mating and spawning at an unusual time.
kost bodlje; A.S. – aksijalni prostor; P. – proces.
If a situation needs a later introduction of the spawning tray, this should be done at least 9 hours after start of the illumination.
B – krajnji dio šipčice peraje.
At this late time of the day, spawning is not induced any longer.
Na vrhu se nalaze Actinotrichia (Act.).
Appendix 4B
Slika 3.
SPAWNING SUBSTRATE FOR FATHEAD MINNOW
Fotografija tijela ribe i prikaz mjesta reza kad je gonada fiksirana u otopini za fiksiranje koja nije 10-postotni neutralni puferirani formalin.
Two or three combined plastic/ceramic/glass or stainless steel spawning tiles and trays are placed in each of the test chamber (e.g. 80 mm length of grey semi-circular guttering sitting on a lipped tray of 130mm length) (see picture).
U tom slučaju ostatak tijela odrezat će se između prednjeg dijela podrepne peraje i anusa s pomoću skalpela (crvena linija); glava s prednjim dijelom ribe stavit će se u otopinu za fiksiranje gonade, a repni dio u 10-postotni neutralni puferirani formalin.
Properly seasoned PVC or ceramic tiles have demonstrated to be appropriate for a spawning substrate (Thorpe et al, 2007).
Papilarni procesi u pravilu se pojavljuju samo u odraslih mužjaka te se nalaze na šipčicama peraja od druge do sedme ili osme od stražnjeg kraja podrepne peraje (slike 1. i 2.).
It is recommended that the tiles are abraded to improve adhesion.
Međutim, procesi se rijetko nalaze na prvoj šipčici peraje od stražnjeg kraja podrepne peraje.
The tray should also be screened to prevent fish from access to the fallen eggs unless the egg adhesion efficiency has been demonstrated for the spawning substrate used.
Ovim standardnim operativnim postupkom (SOP) obuhvaćeno je mjerenje procesa na prvoj šipčici peraje (u ovom SOP-u broj šipčice peraje odnosi se na redoslijed od stražnjeg kraja podrepne peraje).
The base is designed to contain any eggs that do not adhere to the tile surface and would therefore fall to the bottom of the tank (or those eggs laid directly onto the flat plastic base).
PREPORUČENI POSTUPCI ZA UZIMANJE UZORAKA ZA ANALIZU VITELOGENINA
All spawning substrates should be leached for a minimum of 12 hours, in dilution water, before use.
Potrebno je paziti da ne dođe do unakrsne kontaminacije uzoraka VTG-a mužjaka i ženki.
Appendix 5A
Postupak 1.A:
ASSESSMENT OF SECONDARY SEX CHARACTERISTICS IN FATHEAD MINNOW FOR THE DETECTION OF CERTAIN ENDOCRINE ACTIVE CHEMICALS
Debeloglava gavčica, uzimanje krvi iz repne vene/arterije
Overview
Nakon anestezije, korijen repa djelomično se odvaja skalpelom i uzima se krv iz repne vene/arterije s pomoću heparinizirane mikrohematokritske kapilarne cjevčice.
Potentially important characteristics of physical appearance in adult fathead minnows in endocrine disrupter testing include body colour (i.e., light/dark), coloration patterns (i.e., presence or absence of vertical bands), body shape (i.e., shape of head and pectoral region, distension of abdomen), and specialised secondary sex characteristics (i.e., number and size of nuptial tubercles, size of dorsal pad and ovipositor).
Nakon uzimanja krvi plazma se brzo izolira centrifugiranjem pri 15000 g u trajanju od tri minute (ili centrifugiranjem u trajanju od deset minuta pri 15000 g na 4 °C).
Nuptial tubercles are located on the head (dorsal pad) of reproductively-active male fathead minnows, and are usually arranged in a bilaterally-symmetric pattern (Jensen et al.
Postotak hematokrita može se, po želji, utvrditi nakon centrifugiranja.
2001).
Plazma se potom uklanja iz mikrohematokritske cjevčice i pohranjuje u epruvetu za centrifugu uz 0,13 jedinica aprotinina (inhibitora proteaze) na – 80 °C do mogućnosti određivanja VTG-a.
Control females and juvenile males and females exhibit no tubercle development (Jensen et al.
Ovisno o veličini debeloglave gavčice (koja ovisi o spolu), prikupljena količina plazme obično se kreće od pet do 60 mikrolitara po ribi (Jensen et al.
2001).
2001).
There can be up to eight individual tubercles around the eyes and between the nares of the males.
Postupak 1.B:
The greatest numbers and largest tubercles are located in two parallel lines immediately below the nares and above the mouth.
Debeloglava gavčica, uzimanje krvi iz srca
In many fish there are groups of tubercles below the lower jaw; those closest to the mouth generally occur as a single pair, while the more ventral set can be comprised of up to four tubercles.
Druga je mogućnost uzimanje krvi punkcijom srca s pomoću heparinizirane šprice (1000 jedinica heparina po ml).
The actual numbers of tubercles is seldom more than 30 (range, 18-28; Jensen et al.
Krv se stavlja u Eppendorfove epruvete (koje se čuvaju na ledu) i potom centrifugira (pet minuta, 7000 g, na sobnoj temperaturi).
2001).
Plazmu treba premjestiti u čiste Eppendorfove epruvete (u alikvotima ako to dopušta volumen plazme) i odmah smrznuti na – 80 °C do analize (Panter et al.
The predominant tubercles (in terms of numbers) are present as a single, relatively round structure, with the height approximately equivalent to the radius.
1998).
Most reproductively-active males also have, at least some, tubercles which are enlarged and pronounced such that they are indistinguishable as individual structures.
Postupak 2.A:
Some types of endocrine-disrupting chemicals can cause the abnormal occurrence of certain secondary sex characteristics in the opposite sex; for example, androgen receptor agonists, such as 17α-methyltestosterone or 17β-trenbolone, can cause female fathead minnows to develop nuptial tubercles (Smith 1974; Ankley et al.
Japanska medaka, vađenje jetre u medake
2001; 2003), while oestrogen receptor agonists may decrease number or size of nuptial tubercles in males (Miles-Richardson et al.
Uklanjanje ispitne ribe iz ispitne komore
1999; Harries et al.
Ispitnu ribu potrebno je izvaditi iz ispitne komore malom mrežom u obliku žlice.
2000).
Pazite da ne ispustite ispitnu ribu u ostale ispitne komore.
Below is a description of the characterisation of nuptial tubercles in fathead minnows based on procedures used at the U.S. Environmental Protection Agency lab in Duluth, MN.
Ispitne ribe u pravilu je potrebno vaditi sljedećim redoslijedom: kontrola, kontrola s otapalom (ako postoji), najniža koncentracija, srednja koncentracija, najviša koncentracija i pozitivna kontrola.
Specific products and/or equipment can be substituted with comparable materials available.
Osim toga, iz ispitne komore treba izvaditi sve mužjake prije vađenja preostalih ženki.
Viewing is best accomplished using an illuminated magnifying glass or 3X illuminated dissection scope.
Spol ispitnih riba utvrđuje se na temelju vanjskih sekundarnih spolnih obilježja (primjerice oblika podrepne peraje).
View fish dorsally and anterior forward (head toward viewer).
Stavite ribu u spremnik za transport i odnesite je do radne stanice radi vađenja jetre.
Place fish in small Petri dish (e.g. 100 mm in diameter), anterior forward, and ventral down.
Provjerite točnost oznaka na ispitnoj komori i spremniku za transport i provjerite je li broj riba koje su izvađene iz ispitne komore i broj riba preostalih u ispitnoj komori u skladu s očekivanjima.
Focus viewfinder to allow identification of tubercles.
Ako se spol ne može utvrditi iz vanjskog izgleda riba, izvadite sve ribe iz ispitne komore.
Gently and slowly roll fish from side to side to identify tubercle areas.
Spol je u tom slučaju potrebno utvrditi promatranjem gonade ili sekundarnih spolnih obilježja pod stereoskopskim mikroskopom.
Count and score tubercles.
Vađenje jetre
Repeat the observation on the ventral head surface by placing the fish dorsal anterior forward in the Petri dish.
Premjestite ispitnu ribu iz spremnika za transport u otopinu anestetika malom mrežom u obliku žlice.
Observations should be completed within 2 min for each fish.
Nakon anestezije pincetom (običnom) premjestite ispitnu ribu na filtarski papir (ili papirnati ručnik).
Tubercle Counting and Rating
Pri hvatanju ispitne ribe pincetom obuhvatite glavu kako ne bi došlo do kidanja repa.
Six specific areas have been identified for assessment of tubercle presence and development in adult fathead minnows.
Obrišite vodu s površine ispitne ribe filtarskim papirom (ili papirnatim ručnikom).
A template was developed to map the location and quantity of tubercles present (see end of this appendix).
Potom škaricama napravite mali poprečni rez od leđnog dijela vrata do područja srednjeg trbuha.
The number of tubercles is recorded and their size can be quantitatively ranked as: 0- absence, 1-present, 2-enlarged and 3-pronounced for each organism (Fig.
Umetnite škarice u mali rez i zarežite trbuh od točke sa stražnje strane škržnog plašta do prednje strane anusa duž središnje linije abdomena.
1).
Pazite da škarice ne umetnete preduboko kako ne bi došlo do oštećenja jetre i gonade.
Rate 0 — absence of any tubercle.
Obavite sljedeće postupke pod stereoskopskim mikroskopom.
Rating 1 — present, is identified as any tubercle having a single point whose height is nearly equivalent to its radius.
Položite ispitnu ribu trbuhom prema gore na papirnati ručnik (pri ruci moraju biti i staklena Petrijeva zdjelica ili stakalce).
Rating 2 — enlarged, is identified by tissue resembling an asterisk in appearance, usually having a large radial base with grooves or furrows emerging from the centre.
Proširite stijenke trbušne šupljine preciznom pincetom i izvadite unutarnje organe.
Tubercle height is often more jagged but can be somewhat rounded at times.
Prihvatljivo je, po potrebi, i vađenje unutarnjih organa uklanjanjem jedne strane stijenke trbušne šupljine.
Rating 3 — pronounced, is usually quite large and rounded with less definition in structure.
Otkrijte spojeni dio jetre i žučnog mjehura drugom preciznom pincetom.
At times these tubercles will run together forming a single mass along an individual or combination of areas (B, C and D, described below).
Potom uhvatite žučni kanal i odrežite žučni mjehur.
Coloration and design are similar to rating 2 but at times are fairly indiscriminate.
Pazite da ne dođe do trganja žučnog mjehura.
Using this rating system generally will result in overall tubercle scores of < 50 in a normal control male possessing a tubercle count of 18 to 20 (Jensen et al.
Uhvatite jednjak i na isti način razdvojite probavni trakt od jetre.
2001).
Pazite da ne dođe do izlijevanja sadržaja probavnog trakta.
The actual number of tubercles in some fish may be greater than the template boxes for a particular rating area.
Odvojite stražnji dio probavnog trakta od anusa i izvadite ga iz trbušne šupljine.
If this happens, additional rating numbers may be marked within, to the right or to the left of the box.
Uklonite masno tkivo i ostala tkiva s vanjskog dijela jetre.
The template therefore does not need to display symmetry.
Pazite da ne ogrebete jetru.
An additional technique for mapping tubercles which are paired or joined vertically along the horizontal plane of the mouth could be done by double-marking two tubercle rating points in a single box.
Preciznom pincetom uhvatite krvne žile uz jetru i uklonite jetru iz trbušne šupljine.
Mapping regions:
Položite jetru na stakalce.
A — Tubercles located around eye.
Po potrebi, s površine jetre preciznom pincetom uklonite višak masnog tkiva i okolnog tkiva (primjerice trbušnu ovojnicu).
Mapped dorsal to ventral around anterior rim of eye.
Izvažite jetru s pomoću elektroničke analitičke vage i mikroepruvete od 1,5 ml kao tare.
Commonly multiple in mature control males, not present in control females, generally paired (one near each eye) or single in females exposed to androgens.
Zabilježite vrijednost u obrazac (očitanje: 0,1 mg).
B — Tubercles located between nares, (sensory canal pores).
Potvrdite podatke o identifikaciji na naljepnici mikroepruvete.
Normally in pairs for control males at more elevated levels (2- enlarged or 3- pronounced) of development.
Zatvorite poklopac mikroepruvete u kojoj se nalazi jetra.
Not present in control females with some occurrence and development in females exposed to androgens.
Pohranite je u stalak za hlađenje (ili stalak s ledom).
C — Tubercles located immediately anterior to nares, parallel to mouth.
Nakon vađenja jedne jetre instrumente očistite ili ih zamijenite čistima.
Generally enlarged or pronounced in mature control males.
Izvadite jetre svim ribama u spremniku za transport kako je prethodno opisano.
Present or enlarged in less developed males or androgen-treated females.
Nakon vađenja jetara svim ribama u spremniku za transport (odnosno svim mužjacima ili ženkama u ispitnoj komori) stavite sve uzorke jetara u stalak za epruvete s identifikacijskom oznakom i pohranite ih u zamrzivač.
D — Tubercles located parallel along mouth line.
U slučaju uzimanja jetara za preliminarno tretiranje ubrzo nakon vađenja primjerci se do sljedeće radne stanice nose u stalku za hlađenje (ili stalku s ledom).
Generally rated developed in control males.
Nakon vađenja jetre truplo ribe podvrgava se histološkoj pretrazi gonade i mjerenju sekundarnih spolnih obilježja.
Absent in control females but present in androgen-exposed females.
Uzorak
E — Tubercles located on lower jaw, close to mouth, usually small and commonly in pairs.
Pohranite uzorke jetara ispitnih riba na ≤ – 70 °C ako se ne podvrgavaju preliminarnom tretiranju ubrzo nakon vađenja.
Varying in control or treated males, and treated females.
Škarama se načini rez neposredno ispred prsnih peraja.
F — Tubercles located ventral to E. Commonly small and paired.
Škarama se razreže središnja linija abdomena do točke približno 2 mm od kranijalne strane anusa.
Present in control males and androgen-exposed females.
Pincetom se trbušne stijenke šire radi vađenja jetre i ostalih unutarnjih organa.
Tubercle Template:
(Druga je mogućnost bočno pričvrstiti trbušne stijenke).
Numerical Rating
Strelica pokazuje jetru.
Date
Jetra se izrezuje i vadi s pomoću pincete.
resent
Slika 5.
Total Score
Crijeva se nježno povlače pincetom.
nlarged
Slika 6.
ronounced
Škarama se režu oba kraja crijeva i sva mezenterička vezivna tkiva.
Appendix 5B
Slika 7.
ASSESSMENT OF SECONDARY SEX CHARACTERISTICS IN MEDAKA FOR THE DETECTION OF CERTAIN ENDOCRINE ACTIVE CHEMICALS
(ženka) Postupak je jednak za ženke.
Below is a description of the measurement of papillary processes [1], which are the secondary sex characteristics in medaka (Oryzias latipes).
Slika 8.
After the excision of the liver (Appendix 6), the carcass is placed into a conical tube containing about 10 ml of 10 % neutral buffered formalin (upside: head, downside: tail).
Kraj postupka.
If the gonad is fixed in a solution other than 10 % neutral buffered formalin, make a transverse cut across the carcass between anterior region of anal fin and anus using razor, taking care not to harm the gonopore and gonad itself (Fig.3).
Postupak 2.B:
Place the cranial side of the fish body into the fixative solution to preserve the gonad, and the tail side of the fish body into the 10 % neutral buffered formalin as described above.
Japanska medaka (Oryzias latipes), preliminarno tretiranje jetre za analizu vitelogenina
After placing the fish body into 10 % neutral buffered formalin, grasp the anterior region of the anal fin with tweezers and fold it for about 30 seconds to keep the anal fin open.
Uzmite bocu homogenizacijskog pufera iz kompleta za analizu ELISA i ohladite je smrvljenim ledom (temperatura otopine: ≤ 4 °C).
When grasping the anal fin with tweezers, grasp a few fin rays in the anterior region with care not to scratch the papillary processes.
U slučaju upotrebe homogenizacijskog pufera iz sustava EnBio ELISA otopite otopinu na sobnu temperaturu, a potom bocu ohladite smrvljenim ledom.
After keeping the anal fin open for about 30 seconds, store the fish body in 10 % neutral buffered formalin at room temperature until the measurement of the papillary processes (measurement should be conducted after fixing for at least 24 hours).
Izračunajte volumen homogenizacijskog pufera za jetru na temelju njezine mase (dodajte 50 μl homogenizacijskog pufera po mg mase jetre).
Measurement
Primjerice, ako masa jetre iznosi 4,5 mg, volumen homogenizacijskog pufera za jetru iznosi 225 μl.
After fixing the fish body in the 10 % neutral buffered formalin for at least 24 hours, pick up the fish carcass from the conical tube and wipe the formalin on the filter paper (or paper towel).
Sastavite popis volumena homogenizacijskog pufera za sve jetre.
Place the fish abdomen side up.
Priprema jetre za preliminarno tretiranje
Then cut the anal fin using small dissection scissors carefully (it is preferable to cut the anal fin with small amount of pterygiophore).
Neposredno prije tretiranja iz zamrzivača izvadite mikroepruvetu od 1,5 ml u kojoj se nalazi jetra.
Grasp the anterior region of the severed anal fin with tweezers and put it on a glass slide with a several drops of water.
Preliminarno tretiranje jetre mužjaka potrebno je obaviti prije ženki kako bi se izbjegla kontaminacija vitelogeninom.
Then cover the anal fin with a cover glass.
Osim toga, preliminarno tretiranje ispitnih skupina potrebno je obaviti sljedećim redoslijedom: kontrola, kontrola s otapalom (ako postoji), najniža koncentracija, srednja koncentracija, najviša koncentracija i pozitivna kontrola.
Be careful not to scratch the papillary processes when grasping the anal fin with tweezers.
Broj mikroepruveta od 1,5 ml s uzorcima jetara izvađenih iz zamrzivača u određenom trenutku ne smije premašiti broj mikroepruveta koje je moguće istovremeno centrifugirati.
Count the number of the joint plate with papillary processes using the counter under a biological microscope (upright microscope or inverted microscope).
Poredajte mikroepruvete od 1,5 ml s uzorcima jetara prema redoslijedu broja primjerka na stalku s ledom (jetre ne treba otapati).
The papillary processes are recognised when a small formation of processes is visible on the posterior margin of joint plate.
Postupak preliminarnog tretiranja
Write the number of joint plate with papillary processes in each fin ray to the worksheet (e.g. first fin ray: 0, second fin ray: 10, third fin ray: 12, etc.) and enter the sum of this number on the Excel spreadsheet by individual fish.
Dodavanje pufera za homogenizaciju
If necessary, take a photograph of the anal fin and count the number of joint plate with papillary processes on the photograph.
Provjerite popis kako biste utvrdili koji je volumen homogenizacijskog pufera potrebno upotrijebiti za određeni uzorak jetre i podesite mikropipetu (raspon volumena: 100 – 1000 μl) na odgovarajući volumen.
After the measurement, put the anal fin into the conical tube described in (1) and store it.
Na mikropipetu stavite čisti vrh.
Fig.1.
Uzmite homogenizacijski pufer iz boce s reagensom i dodajte pufer u mikroepruvetu od 1,5 ml u kojoj se nalazi jetra.