Kugadzira girafu rekuyeranisa bhuruu yemethylene: sampuli dzisina dhayi, muganhu, uye kutwasuka kwemutsara

Vaongorori vaviri vanoshandisa bhodhoro rimwe chete remhinduro yekutanga yebhuruu yemethylene, uye mumwe nomumwe anogadzira girafu rekuyeranisa. Wokutanga anowana mutsara wakatwasuka unopfuura panosangana zero dzematemo egirafu uye anoyera sampuli dzine kusangana kusingazivikanwi nechivimbo. Wechipiri anowana mutsara unokotama kumusoro, panochekwa demo risiri pedyo nezero, uye mhedzisiro yesampuli inoshanduka pakunyungudutswa. Dhayi imwe chete, mudziyo mumwe chete, kureba kwemafungu kumwe chete. Musiyano hausi unyanzvi. Magadzirirwo acho: zvakaiswa musampuli isina dhayi, kusangana kwakasarudzwa, uye kana muganhu wakaongororwa wakaramba uri munharaunda umo modhi imwe iri nyore ichiri kushanda.

Girafu rekuyeranisa chibvumirano pakati pemhinduro yemudziyo nekusangana kwechinhu, chinoshanda chete pasi pemamiriro arakagadzirwa maari. Kubhuruu yemethylene, chibvumirano ichocho chine nzvimbo isina kusimba inozivikanwa. Mamorekuru edhayi anobatana mumvura, saka mhando yemamorekuru inotora chiedza inoshanduka nekusangana. Girafu rakagadzirwa richiyambuka shanduko iyoyo harizotwasuki, uye hapana ongororo yenhamba inogona kurigadzirisa. Chinyorwa chino chinoratidza magadzirirwo egirafu kuitira kuti mutsara uve nerevo: sarudza sampuli isina dhayi yakakodzera, chengeta muganhu munharaunda inotongwa nemonoma, dzokorora zviyero nenzira yakatendeseka, uye dudzira kukotama semashoko anobatsira kwete semheremhere yezviyero.

Tanga nesampuli isina dhayi, kwete nezviyero

Sampuli isina dhayi ndiyo inotsanangura zero, uye kubhuruu yemethylene inofanira kuva nezvinhu zvose kunze kwedhayi. Mvura yegiredhi rimwe chete, mushonga unochengetedza pH wakafanana nepH yakafanana, munyu wakafanana, chikamu chakafanana chemumwe munyungudutsi wakasanganiswa, uye tembiricha yakafanana. Gadzira mhinduro dzekuyeranisa mumusanganiswa iwoyo chaiwo. Chikonzero chacho ndechemakemikari, kwete chemitemo yebasa. Munyu nemunyungudutsi zvinoshandura kuenzana pakati pemonoma, daima, nemapoka makuru emamorekuru akabatana, izvo zvinoshandura spectrum pakureba kwemafungu kuri kuyerwa. Sampuli ine mvura chete isina dhayi haigoni kugadzirisa shanduko yemhando dzemamorekuru yakakonzerwa nemusanganiswa. Ongororo yakazara yemaspectrum ebhuruu yemethylene mumvura, inowanika kuburikidza nerekodhi yayo inoverengwa pachena, yakashandura sodium chloride kubva pa0 kusvika pa0.15 M nokuti simba remaioni rinoshandura kuenzana uku, uye kubatana kwemamorekuru kwakawedzera nemunyu.

Mugumisiro mumwe unoshanda: usamboyeranisa mumvura yakabviswa maioni wozoyera sampuli mumushonga wephosphate unochengetedza pH kana mumvura yetsvina yapfuura nepasefa. Kana pasina musanganiswa usina chinhu chiri kuongororwa, kuwedzera chiyero chinozivikanwa musampuli yacho kuri nani pane kushandisa girafu rekunze rine musanganiswa usina kufanana. Nhungamiro ine chokuita nokuchengetedza kuvimbika kweongororo dzemumvura iri muhurukuro yemusanganiswa nezvinovhiringidza, uye misimboti yakajairika yeongororo dzinoyera huwandu hwedhayi inotsanangurwa muchidimbu chemagadzirirwo eongororo.

Sarudza muganhu unotongwa nemhando imwe yemamorekuru

Kuyera huwandu pabhandi remafungu marefu riri pedyo ne664 nm kunofungidzira kuti dhayi zhinji iri muchimiro chemonoma. Mumvura isina zvimwe zvakawedzerwa, fungidziro iyoyo inoshanda chete kana mhinduro dzichiri dzakanyungudutswa, kazhinji munharaunda yakaderera yemicromolar. Materu akaburitswa kubva pazviyero anoratidza panogumira izvi: kuyeranisa kunochengetwa pasi peanenge 7 micromolar mumasero e1 cm kunopa mutserendende uri pedyo ne82,000 M-1 cm-1, ukuwo kuyeranisa kunotambanudzwa kusvika paanenge 37 micromolar kuchidzikira kusvika paanenge 67,100 nokuti zviyero zvepamusoro zvinenge zvatova nedaima yakawanda inooneka. Koefishenti imwe neimwe yataurwa pano ine mamiriro ayo akanyorwa mugwaro remaspectrum ebhuruu yemethylene, inova nzvimbo yakakodzera kuenzanisa nhamba; chinyorwa chino chinodzishandisa chete kusarudza muganhu.

Mitemo yemagadzirirwo inobva pane izvi zvakananga:

1. Chengeta chiyero chepamusoro chakanyungudutswa zvakakwana kuti chimiro chespectrum chisachinja. Paunenge uchigadzira nzira, ongorora nharaunda yose yechiedza chinooneka (inenge 550 kusvika 750 nm) pachiyero chimwe nechimwe pane kuverenga kureba kwemafungu kumwe chete. Kana maspectrum agadziridzwa kuti aenzaniswe achichinja chimiro nekusangana, mamiriro emakemikari ari kuchinja uye muganhu wakafara zvikuru.
2. Tevera reshiyo yemwero wekutorwa kwechiedza pedyo ne610 nm kumwero wekutorwa kwechiedza pedyo ne665 nm pazviyero zvose. Reshiyo inokwira nekusangana inoreva kuti kabandiko kadaima kari kukura uye modhi yemonoma iri kutadza. Chiratidzo ichi chinotsanangurwa zvakadzama munhungamiro yezvinoitika pakubatana kwemamorekuru nekusangana.
3. Shandisa sero re1 cm pabasa rekusangana kwakaderera kwemicromolar. Kana mutserendende uri pedyo ne82,000, 5 micromolar inopa mwero wekutorwa kwechiedza uri pedyo ne0.41 uye 10 micromolar inopa anenge 0.82, zvose zviri mukati memiganhu inoshanda zvakanaka kumidziyo mizhinji.
4. Pamhinduro dzekutanga dzine kusangana kwakanyanya, pfupisa nzira yechiedza pane kubvuma mwero wekutorwa kwechiedza uri pamusoro zvikuru pe1, asi yeuka kuti nzira pfupi inogadzirisa kuyerwa kwechiedza, kwete mamiriro emakemikari. Kubatana kwemamorekuru kunoenderana nekusangana, kwete kureba kwenzira yechiedza, saka mhinduro yekutanga ine kusangana kwakanyanya ichiri kuda nzira yayo inofunga nezvemhando dzemamorekuru, kwete girafu remonoma yakanyungudutswa.

Kuenzanisa kwemodhi kwakazara kunotsigira zvakawanda zviri pano kwakabatanidza maspectrum 224 kubva pa1.1 micromolar kusvika pa3.4 millimolar, uye kwakawana kuti chiratidzo chetetramera chaigona kutoonekwa pasi pe34 micromolar. Zvakawanikwa izvi zvinopa yambiro: nharaunda ine mutsara wakatwasuka yakatetepa kupfuura zvinofungidzirwa, saka isimbise pane kungoifungidzira.

Zviyero, kudzokorora, uye kutevedzana

Girafu rine mapoinzi matatu hariratidzi chinhu. Nhungamiro yekusimbisa nzira iri muICH Q2(R2) inoda kusangana kungangosvika kushanu kuti kutwasuka kwemutsara kuongororwe, uye zviyero zvitanhatu kusvika zvisere zvisiri zero zvinopa girafu rebasa rinogutsa zvikuru, kunyanya pedyo nepamusoro panotanga kubatana kwemamorekuru. Rongedza zviyero kuti zvifukidze muganhu wesampuli waunoda, uchiwedzera zviyero pedyo nepanonyanya kutarisirwa kukotama, kwete munhevedzano yejometri yakasarudzwa nokuti iri nyore.

Yera chiyero chimwe nechimwe kaviri kana katatu, uchichigadzira patsva chakazvimirira pane kuverenga katatu kubva mufurasiki rimwe chete. Zviyero zvinodzokororwa zvichigadzirwa zvakazvimirira zvinoburitsa zvikanganiso zvekugadzira; kuverenga kakawanda kubva mufurasiki rimwe chete kunongoburitsa mheremhere yemudziyo. Ongororo yakazara ya2020 yakayera mhinduro imwe neimwe katatu, izvo zvichiri nzira yakanaka yokutangira. Ita kuti kutevedzana kwekuverenga kuve kusingateveri hurongwa hwakasimba kuitira kuti kushanduka kwezviyero nenguva, kunamira pagirazi, kana kuchinja kwetembiricha zvisazvitaridza sekukotama. Ongororo yemukombe wekuyerera chiedza wequartz yakawana kuti inenge 3 percent yedhayi yakanamira kubva mumhinduro ye34 micromolar mukati me10 minutes, saka chengeta nguva yekubatana, mukombe wekuyerera chiedza, kuchenesa, nekutevedzana zvakafanana panguva yose yekuyera.

Muenzaniso wakaverengwa wokuratidza nzira (hausi mashoko akayerwa)

Nhamba dziri pasi apa dzakagadzirwa kuti dziratidze kuverenga. Hadzisi zviyero zvemurabhoritari uye hadzifaniri kutaurwa semashoko ebhuruu yemethylene. Chinokosha imaitiro ekuongorora anodzienderedza.

Ngatitii pane zviyero zvitanhatu mumushonga wakafanana unochengetedza pH, nzira yechiedza ye1 cm, zvichiverengwa pa665 nm zvichienzaniswa nesampuli isina dhayi ine musanganiswa wakafanana:

Kuenzanisa mutsara wakatwasuka neavhareji yemwero wekutorwa kwechiedza zvichienderana nekusangana kunopa mutserendende uri pedyo ne0.0805 mayuniti ekutorwa kwechiedza pamicromolar uye panochekwa demo pari mukati memayuniti mashoma echuru chimwe cheyuniti yekutorwa kwechiedza kubva pazero. Kugovanisa mutserendende nekureba kwenzira yechiedza kunoushandura kuita koefishenti inooneka yekutorwa kwechiedza: 0.0805 pamicromolar yakaenzana neanenge 80,500 M-1 cm-1, zvinoenderana nemuganhu unotongwa nemonoma mumvura. Zvinoongororwa zvinokosha haisi R squared, iyo icharatidzika yakanaka zvikuru chero zvodii. Ndipo panochekwa demo pedyo nezero, reshiyo yeA610/A665 isinganyanyi kuchinja kubva paS2 kusvika paS5, uye misiyano yasara yakapararira isina maitiro anotevedzana. Kukwira kuduku kwereshiyo paS6 chiratidzo chaicho chokuti chiyero ichocho chibviswe kana kuti 7 micromolar imakwe semuganhu wepamusoro wenguva ino wemusanganiswa uyu pamwe nemudziyo uyu.

Sampuli ine kusangana kusingazivikanwi yakaverengwa pa0.245 pasi pemamiriro akafanana inoshandurwa zvakananga: 0.245 yakagovaniswa ne0.0805 pamicromolar inopa anenge 3.0 micromolar, kusati kwashandiswa chero chinhu chinogadzirisa kunyungudutswa kwesampuli. Taura mhedzisiro pamwe nemamiriro ayo: kureba kwemafungu, nzira yechiedza, musanganiswa wesampuli isina dhayi, muganhu wekuyeranisa, uye zuva.

Pepa rekushandira rekuyeranisa rinogona kuzadzwa

Kopa muenzaniso uyu murekodhi yerabhoritari pamusanganiswa mutsva mumwe nomumwe. Musanganiswa mumwe, pepa rimwe, girafu rimwe.

Mutsara wechimiro chereagent une chikonzero chokuva ipapo. Bhuruu yemethylene inotengeswa inogona kuva munyu usina mvura yakabatanidzwa kana ine mvura yakabatanidzwa yakadai setrihydrate, uye huremu hwemolar husiri ihwo hunokanganisa kusangana kwose kwakarongwa pasina kuzvitaridza. Kuongororwa kwekuchena uye zvinodiwa pazvinhu zvinotangiswa kunotsanangurwa munhungamiro yeunhu negiredhi.

Simbisa nechiyero chekuongorora chakazvimirira

Girafu rekuyeranisa harifaniri kuzviongorora roga. Mushure mekuenzanisa mutsara, gadzira chiyero chimwe chekuongorora kubva mumhinduro yekutanga yakayerwa huremu yakazvimirira, chine kusangana kuri pedyo nepakati pemuganhu wakagamuchirwa, wochiverenga sechisina kusangana kunozivikanwa. Kubvumirana mukati memaperesenti mashoma kunosimbisa nhanho dzekugadzira; kusabvumirana kunonongedzera kumhinduro yekutanga kana kunyungudutswa pane kunongedzera kumudziyo. Dzokorora chiyero chekuongorora nechikwata chimwe nechimwe chesampuli. Kana chichishanduka nenguva girafu risina kuchinja, fungidzira mhinduro dziri kushandiswa kana mukombe wekuyerera chiedza usati wafungidzira sampuli. Tsika iyi imwe chete inobata zvikanganiso zvizhinji zvisingazvitaridzi: huremu hwemolar hwechinhu chine mvura yakabatanidzwa hwakashandiswa pabhodhoro rechinhu chisina mvura iyoyo, micropipette isina kuyeraniswa zvakarurama, kana mushonga unochengetedza pH wakagadzirwa nesimba remaioni risiri iro. Nyora mugumisiro wakawanikwa uchienzaniswa newaitarisirwa pachiyero chekuongorora papepa rekushandira pedyo nemutserendende, kuitira kuti anozoverenga aone kuti mutsara wakaedzwa, kwete kungodhirowewa.

Kududzira kusatwasuka kuti uone chikonzero

Kana girafu rikakotama, chimiro chekukundikana chinoratidza chikonzero. Kukotamira pasi pamwero mukuru wekutorwa kwechiedza pasina kuchinja kwechimiro chespectrum kunonongedzera kumudziyo, kazhinji chiedza chakatsauka chichiita kuti mwero mukuru wekutorwa kwechiedza uverengwe uri pasi pane chaiwo. Kukotama kunoperekedzwa nokukura kwekabandiko ke610 nm kunonongedzera kumakemikari: daima nemapoka makuru emamorekuru akabatana zvinotora chiedza zvakasiyana nemonoma. Sampuli isina dhayi isingasviki pazero, kana zviyero zvinodzikira nekufamba kwenguva, zvinonongedzera kumudziyo wakabata mhinduro: kunamira pagirazi kana pamikombe yekuyerera chiedza, kunyanya kana nguva dzekubatana dzakasiyana pakutevedzana kwekuyera. Uye mutsara wakanaka unopfuura pamapoinzi asi uchiri kupa mhedzisiro yesampuli isina kururama unonongedzera kumusanganiswa: sampuli dzakasiyana nemhinduro dzekuyeranisa, saka gadzirazve girafu mumusanganiswa wesampuli kana kushandisa kuwedzera zviyero zvinozivikanwa musampuli. Dhirowa misiyano yasara zvichienderana nekusangana nguva dzose. Kupararira kusina maitiro anotevedzana kunotsigira modhi yemutsara wakatwasuka; chimiro chinokotamira kumusoro kana pasi chinoiramba nenzira yakavimbika kupfuura chero muganhu weR squared, pfungwa inotsigirwa pachena nenhungamiro yekusimbisa nzira.

Bhuruu yemethylene inopa mhedzisiro yakanaka kana kurongeka uku kuchitevedzwa. Bhandi remonoma yayo rakasimba, mamiriro ayo emakemikari akanyatsotsanangurwa, uye nzira dzainokundikana nadzo dzinozviratidza muspectrum dzisati dzakanganisa nhamba. Gadzira girafu rine muganhu mutete, shandisa sampuli isina dhayi yakakodzera, dzokorora zviyero zvakazvimirira, uye rega chimiro chemaspectrum chitungamirire kusarudzwa kwemuganhu. Mutsara unozobuda mupfupi, asi poinzi imwe neimwe pauri ndeyechokwadi.

Kuverenga kwokuratidza nzira muchinyorwa chino kunoshandisa nhamba dzakagadzirwa kuti kuratidze mashandiro acho. Makoefishenti ereferensi akayerwa ane mamiriro awo ose anowanikwa mugwaro remaspectrum rakabatanidzwa, uye ongororo yekubatana kwemamorekuru inotsigira izvi yakanyorwa paPMID 33251415.