Kupaka utoto wa methylene blue mosavuta poyerekeza ndi kupaka utoto kwa Gram: zimene njira iliyonse ingatsimikizire

Wophunzira amakonza chopaka cha maselo kuchokera ku mabakiteriya okulitsidwa omwe alembedwa kuti ndi E. coli, amathira methylene blue kuti aphimbe chopakacho kwa mphindi imodzi, amachitsuka, kenako amachiyang'ana ndi lensi yogwiritsa ntchito mafuta. Maselo abuluu ooneka ngati timitengo amadzaza malo oonera. Lipoti la labotale limati: "Maselo ooneka ngati timitengo a Gram-negative azindikiridwa kuti ndi E. coli." Zimene waona n'zolondola. Koma zimene watsimikiza sizikuthandizidwa ndi umboni wa pa galasi limenelo.

Cholakwikacho chili pa kumvetsa mfundo, osati pa luso lochitira njirayo. Wophunzirayo watenga kupaka utoto mosavuta ngati kuti ndi mayeso osiyanitsa maselo. Kupaka utoto mosavuta ndi kupaka utoto kwa Gram kumayankha mafunso osiyanasiyana chifukwa zimatsatira mfundo zosiyanasiyana za kupaka utoto. Njira imodzi imafunsa kuti: pali mawonekedwe otani? Inayo imafunsa kuti: khoma la selo limachita bwanji likayesedwa? Mukasokoneza njira ziwirizi, galasi lopakidwa utoto molondola lingatulutse lipoti losathandizidwa ndi umboni.

Kuyerekezeraku kumasiyanitsa njira ziwirizi, kumasonyeza zimene zotsatira za iliyonse zimakulolani kunena, ndipo kumayankha mafunso awiri omwe ophunzira amafunsa kwambiri: kodi methylene blue ingagwiritsidwe ntchito popaka utoto kwa Gram, ndipo kupaka E. coli utoto wa methylene blue kumasonyeza chiyani kwenikweni?

Yankho lachidule

Kupaka utoto wa methylene blue mosavuta kumapangitsa selo lililonse lomwe lingalandire utoto kukhala labuluu mofanana. Choncho kumasonyeza mawonekedwe (timitengo, mipira, zopindika ngati kasupe) ndi kakonzedwe ka maselo (pawokha, awiriawiri, m'unyolo), koma sikumasonyeza gulu la Gram. Kupaka utoto kwa Gram kumawonjezera ayodini, sitepe yochotsa utoto, ndi utoto wachiwiri, choncho kumagawa mabakiteriya kukhala Gram-positive (ofiirira) ndi Gram-negative (apinki kapena ofiira), potengera mmene zigawo zawo zakunja zimasungira utoto. Maselo abuluu ooneka ngati timitengo mutagwiritsa ntchito methylene blue amatanthauza kuti "pali maselo ooneka ngati timitengo." Maselo apinki ooneka ngati timitengo mutamaliza njira yonse ya Gram amatanthauza kuti "pali maselo ooneka ngati timitengo a Gram-negative." Mawu achiwiriwa ali ndi umboni wamphamvu kwambiri chifukwa njirayo yachita mayeso owonjezera.

Kuti mudziwe zambiri za gulu lonse la njira zopaka mabakiteriya utoto umodzi zomwe nkhaniyi ikukhudza, onani buku la zithunzi la kupaka utoto poyang'ana ndi maikulosikopu.

Kupaka utoto mosavuta: utoto umodzi, funso limodzi

Methylene blue ndi utoto wa mtundu wa basic (wokhala ndi ayoni ya chaji yabwino). Ayoni yake ya utoto imakhala ndi chaji yabwino ndipo imamangirira ku zigawo za selo zokhala ndi chaji yoipa, kuphatikizapo ma nucleic acid ndi magulu a asidi a pamwamba pa selo. Choncho maselo amaoneka abuluu pomwe kumbuyo kwake kumaoneka kotumbululuka. OpenStax imafotokoza mfundo imeneyi yopaka maselo utoto m'chaputala chake cha kupaka utoto. Ayoni yabwino ya utotoyo ndi ayoni yaing'ono, yathyathyathya ya phenothiazine yomwe yafotokozedwa mwatsatanetsatane mu nkhani ya kapangidwe ndi katundu wa mankhwala a methylene blue; ayoni ya chloride yokhala ndi chaji yotsutsana nayo siimene imapereka mtundu.

Njira yodziwika yophunzitsira imasiyidwa yosavuta mwadala: pangani chopaka chopyapyala cha maselo, chisiyeni chiume ndi mpweya, chimangirireni pa galasi pogwiritsa ntchito kutentha, chiphimbeni ndi methylene blue wosungunuka m'madzi wa pafupifupi 1% kwa pafupifupi mphindi imodzi, chisambitseni modekha ndi madzi, chiumitseni pochikhudza ndi pepala loyamwa, ndipo chiyang'aneni ndi lensi yogwiritsa ntchito mafuta. Njira yophunzitsira ya LibreTexts imapereka ndendende njira imeneyi ya 1% kwa mphindi imodzi, ndipo imanena kuti malabotale ena amawonjezera nthawi. Palibe chothandizira kumangirira utoto, palibe chochotsa utoto, ndipo palibe utoto wachiwiri. Kusakhalapo kwa zimenezi ndi mfundo yaikulu. Pakakhala utoto umodzi wokha ndipo palibe kuchotsa utoto mosankha, selo lililonse lomwe limamangirira utotowo limakhala la mtundu womwewo.

Pa E. coli, zimene zikuyembekezeka kuoneka ndi maselo abuluu ooneka ngati timitengo (bacilli), pawokha, awiriawiri, kapena m'magulu ang'onoang'ono. Mtundu wabuluu umachokera ku utoto. Mawonekedwe ngati timitengo amachokera ku chamoyochacho. Koma kuphatikizana kwa zimenezi sikuzindikiritsa chamoyochacho, chifukwa mitundu yambiri ya mabakiteriya imaoneka ngati timitengo ndipo ambiri amapaka buluu akagwiritsa ntchito njira yosavuta ya utoto. ATCC imanena momveka bwino za malire amenewa a kungosonyeza mawonekedwe m'buku lake lotsogolera kukulitsa tizilombo tating'ono. Lipoti lolondola limati: "Maselo abuluu ooneka ngati timitengo aonedwa." Chilichonse chopitirira mawonekedwe ndi kakonzedwe ka maselo chimafuna umboni wina.

Kupaka utoto kwa Gram: zinthu zinayi, mayeso amodzi a khalidwe la maselo

Kupaka utoto kwa Gram ndi njira yosiyanitsa maselo. Maselo onse amadutsa m'masitepe omwewo, ndipo amaonedwa kuti ndi ati omwe amasunga utoto woyamba ndi ati omwe amautaya. Njira za kupaka utoto kwa Gram za American Society for Microbiology zimafotokoza dongosolo lokhazikika la masitepe anayi: crystal violet (utoto woyamba), ayodini ya Gram (chothandizira kumangirira utoto), mowa kapena mowa wosakanizidwa ndi acetone (chochotsa utoto), kenako safranin (utoto wachiwiri).

Kafotokozedwe kodziwika pophunzitsa kamafotokoza kusiyanaku potengera kapangidwe ka zigawo zakunja za selo. Maselo a Gram-positive ali ndi khoma lokhuthala la peptidoglycan lomwe limataya madzi ndikusunga gulu lolumikizana la crystal violet ndi ayodini posambitsa ndi mowa. Maselo a Gram-negative ali ndi wosanjikiza wopyapyala wa peptidoglycan komanso nembanemba yakunja yokhala ndi mafuta ambiri yomwe chochotsa utoto chimasokoneza. Choncho gulu lolumikizanalo limatsukidwa kutuluka, ndipo maselowo amalandira safranin. Nkhani yowunikira kupaka utoto kwa Gram ya NCBI Bookshelf imafotokoza nkhani imeneyi ya khoma la selo. Akatswiri amawonjezera kuti kufotokoza zonse potengera fiziki ndi kemistri n'kovuta kuposa kungoyang'ana kukhuthala kwa khoma kokha, koma pomasulira zotsatira mu labotale, lamulo la khalidweli limagwira ntchito: wofiirira amatanthauza kuti utoto wasungidwa, ndipo wapinki amatanthauza kuti utoto woyamba watayika kenako selo lapakidwa utoto wachiwiri.

Zithunzi zofotokozera. Ndi zojambula zoyerekezera njira, osati zithunzi za kamera zojambulidwa kudzera pa maikulosikopu. Mitunduyo yasiyanitsidwa mosavuta kuti iphunzitse mfundo zake: mtundu umodzi popaka utoto mosavuta, mitundu iwiri pambuyo pa kuchotsa utoto kwa Gram.

Kodi methylene blue ingagwiritsidwe ntchito popaka utoto kwa Gram

Funso limeneli lingamveke m'njira ziwiri, ndipo njira zimenezi zimafuna mayankho osiyanasiyana.

Tanthauzo loyamba: kodi kuthira methylene blue kamodzi kungalowe m'malo mwa njira yonse ya Gram? Ayi. Popanda ayodini, chochotsa utoto, ndi utoto wachiwiri, palibe sitepe yosiyanitsa maselo, choncho palibe gulu la Gram lomwe layesedwa. Malo oonera abuluu mutagwiritsa ntchito methylene blue yokha ndi zotsatira za kupaka utoto mosavuta, ngakhale botolo lake litakhala pafupi ndi zinthu za Gram. Musalembe zimenezi ngati zotsatira za Gram.

Tanthauzo lachiwiri: kodi methylene blue ingalowe m'malo mwa safranin monga utoto wachiwiri pa sitepe 4? Potengera ntchito yake, ingapake maselo amene achotsedwa utoto, ndipo ntchito yophunzitsira ya m'buku imaneneratu kuti kusinthaku kungapereke maselo a Gram-positive ofiirira ndi maselo a Gram-negative abuluu. Mfundoyi n'yomveka chifukwa maselo a Gram-negative amene achotsedwa utoto molondola amakhala pafupifupi opanda utoto ndipo amalandira utoto wina wokhala ndi ayoni ya chaji yabwino. Koma imeneyi si njira yokhazikika. Njira za ASM zimatchula safranin, pamodzi ndi zinthu zina zovomerezeka zolowa m'malo mwake monga basic fuchsin m'njira zina, osati methylene blue. Palinso vuto logwiritsa ntchito: kusiyanitsa buluu ndi wofiirira n'kovuta kuposa kusiyanitsa pinki ndi wofiirira, makamaka kwa oyamba kumene ndiponso pamene nyali za maikulosikopu zafooka. Pa maphunziro ndi pa zotsatira zilizonse zofunika, gwiritsani ntchito utoto wachiwiri umene njira yanu yatchula, nthawi zambiri safranin, ndipo tengani kugwiritsa ntchito methylene blue ngati utoto wachiwiri kukhala njira yosinthidwa yowonetsera mfundo, osati njira yofanana ndi yokhazikika.

Funso lofanana la mmene methylene blue imayerekezedwera ndi mitundu ina ya utoto umodzi likufotokozedwa mu kuyerekeza kwa njira zosavuta zopaka mabakiteriya utoto.

Zimene kupaka E. coli utoto wa methylene blue kumatsimikizira, ndi zimene sikumatsimikizira

E. coli ndi bakiteriya wooneka ngati kamtengo wa Gram-negative, koma chiganizo chimenechi chimaphatikiza mfundo zochokera ku njira zingapo: mawonekedwe kuchokera ku kuyang'ana ndi maikulosikopu, khalidwe la Gram kuchokera ku kupaka utoto kwa Gram, ndi kudziwika kwa mtundu kuchokera ku kukulitsa mabakiteriya pamodzi ndi mayeso a biochemistry kapena a mamolekyulu. Chopaka cha methylene blue chimangopereka gawo loyamba lokha. Chimatsimikizira kuti pali maselo ooneka ngati timitengo m'chitsanzo chopakidwa utoto ndi mmene akonzedwera. Sichimatsimikizira khalidwe la Gram, chifukwa palibe mayeso ochotsa utoto omwe achitika. Sichizindikiritsa mtundu wa mabakiteriya, chifukwa maselo ooneka ngati timitengo a mitundu yambiri amafanana pa mlingo umenewu wa kuonetsa tsatanetsatane.

Apa ndi pamene malipoti amalakwika m'njira yodziwikiratu. Wolemba amalowetsa chidziwitso chomwe anali nacho kale ("mabakiteriya okulitsidwawa akuyenera kukhala E. coli, ndipo E. coli ndi Gram-negative") mu mzere wa zimene waona. Choncho galasilo limaoneka ngati likutsimikizira zimene kwenikweni zinangoganiziridwa. Lipoti lomveka bwino limasiyanitsa mbali zimenezi: "Kupaka utoto wa methylene blue mosavuta: maselo abuluu ooneka ngati timitengo aonedwa; mawonekedwe okha, palibe gulu la Gram lomwe layesedwa." Kenako, ngati kupaka utoto kwa Gram kwachitika molondola pogwiritsa ntchito zitsanzo zoyerekezera: "Kupaka utoto kwa Gram: maselo apinki ooneka ngati timitengo, ogwirizana ndi Gram-negative." Kudziwika kwa mtundu wa mabakiteriya kumabwera pambuyo pake, kuchokera ku njira yozindikiritsa, osati kuchokera ku mtundu wa utoto wokha.

Kalozera womasulira zotsatira mu labotale

Gwiritsani ntchito lamulo ili posankha momwe mungatanthauzire zotsatira mu labotale. Ngati maselo onse ali ndi mtundu umodzi ndipo njirayo yagwiritsa ntchito utoto umodzi popanda chochotsa utoto, mwachita kupaka utoto mosavuta. Lembani mawonekedwe, kuyerekezera kwa kukula, ndi kakonzedwe ka maselo. Lekezani pomwepo. Ngati njirayo yagwiritsa ntchito crystal violet, ayodini, chochotsa utoto, ndi utoto wachiwiri pamodzi ndi zitsanzo zoyerekezera zodziwika za Gram-positive ndi Gram-negative, mwachita kupaka utoto kwa Gram. Lembani mtundu wa utoto pamodzi ndi mawonekedwe: maselo ozungulira ofiirira m'unyolo, maselo apinki ooneka ngati timitengo, ndi zina zotero.

Zolakwika zitatu ndizimene zimayambitsa chisokonezo chambiri. Choyamba, kutanthauzira mopitirira umboni kuti maselo abuluu ooneka ngati timitengo amatsimikizira E. coli. Chachiwiri, kulemba galasi la methylene blue pansi pa mutu wakuti "Kupaka utoto kwa Gram" m'kope, zomwe owerenga pambuyo pake angatanthauzire molakwika. Chachitatu, kusintha utoto wachiwiri mosasamala kenako kuyerekezera mitundu pakati pa malabotale ngati kuti mitundu yogwiritsidwa ntchito inali yofanana. Cholakwika chilichonse chili ndi njira yomweyo yochikonza: tchulani njira imene yachitikadi, ndipo nenani zokhazo zimene njirayo imayesa.

Kupaka utoto mosavuta kuli ndi ntchito yake makamaka chifukwa kumatsimikizira zochepa. N'kofulumira, kumalolera zolakwika zazing'ono, ndipo n'kokwanira pamene funso likukhudza mawonekedwe: kodi pali chilichonse, ndipo chili ndi mawonekedwe otani? Pamene funso likukhudza kugawa m'magulu, chitani mayeso osiyanitsa maselo. Galasi labuluu limakuuzani mmene maselo amaonekera. Galasi lofiirira kapena lapinki limakuuzani mmene makoma awo amachitira. Palibe galasi lililonse mwa awiriwa lomwe, palokha, limakuuzani mtundu wa mabakiteriya.