Ukufaka umbala olula ngemethylene blue xa kuthelekiswa nokufaka umbala ngeGram: oko indlela nganye inokukungqina

Umfundi ulungiselela umqatyiso kumkhuliso obhalwe E. coli, awugqume ngemethylene blue umzuzu omnye, awuhlambulule, aze ajonge ngelensi yokuntywiliselwa kwioyile. Iiseli ezizintonga eziluhlaza zigubungela indawo ebonakalayo. Ingxelo yelebhu ithi: "Iibhaktheriya ezizintonga zeGram-negative zichongwe njenge-E. coli." Oko kubonwayo kuchanekile. Kodwa eso silayidi asisixhasi eso sigqibo.

Impazamo isekuqondeni ingcamango, kungekhona ekwenzeni umsebenzi. Umfundi uthathe ukufaka umbala olula ngokungathi luvavanyo lokwahlula. Ukufaka umbala olula nokufaka umbala ngeGram kuphendula imibuzo eyahlukileyo kuba kusekelwe kwiingcamango ezahlukileyo zokufaka umbala. Enye ibuza ukuba: yeyiphi imilo ekhoyo. Enye ibuza ukuba: udonga lweseli luziphatha njani xa luvavanywa. Ukuba ezi zimbini ziyabhidaniswa, isilayidi esifakwe umbala ngokuchanekileyo sinokukhokelela kwingxelo engenabungqina.

Olu thelekiso lugcina ezi ndlela zimbini zahlukene, lubonise ukuba isiphumo ngasinye sikuvumela ukuba ubange ntoni, luze luphendule imibuzo emibini abafundi abayibuza kakhulu: ingaba imethylene blue ingasetyenziswa ekufakeni umbala ngeGram, yaye ukufaka umbala ngemethylene blue kwi-E. coli kubonisa ntoni ngokwenene.

Impendulo emfutshane

Ukufaka umbala olula ngemethylene blue kunika yonke iseli enokufakwa umbala umbala oluhlaza ofanayo, ngoko kubonisa imilo (iintonga, iibhola, iimilo ezijijekileyo) nolungelelwaniso (nganye, ngazimbini, amatyathanga), kodwa akubonisi nto ngodidi lweGram. Ukufaka umbala ngeGram kongeza i-iodine, inyathelo lokususa umbala, nedayi yesibini, ngoko kwahlula iibhaktheriya zibe yiGram-positive (ezimfusa) neGram-negative (ezipinki okanye ezibomvu) ngokwendlela izigqubuthelo zazo eziyigcina ngayo idayi. Iiseli ezizintonga eziluhlaza emva kwemethylene blue zithetha ukuba "kukho iiseli ezimile okweentonga." Iiseli ezizintonga ezipinki emva kolandelelwano olupheleleyo lweGram zithetha ukuba "kukho iiseli ezizintonga zeGram-negative." Ingxelo yesibini inobungqina obomeleleyo kuba inkqubo yenze uvavanyo olongezelelweyo.

Ukuze ufunde ngeqela elibanzi leendlela zedayi enye zokufaka umbala kwiibhaktheriya elikulo eli nqaku, jonga iatlasi yokufaka umbala kwimicroscopy.

Ukufaka umbala olula: idayi enye, umbuzo omnye

Imethylene blue yidayi esisiseko (yecation). I-ion yayo enemibala inentlawulo yombane evumayo kwaye ibophelela kwizakhiwo zeseli ezinentlawulo engavumiyo, kuquka ii-asidi zenucleic namaqela aneasidi kumphezulu, ngoko iiseli zibonakala ziluhlaza kumgangatho ongemva ombatshileyo. I-OpenStax ichaza lo mgaqo wokufaka umbala kwiiseli kwisahluko sayo sokufaka umbala. Ication yedayi ngokwayo yi-ion encinane, esicaba yephenothiazine echazwe ngokweenkcukacha kwisikhokelo sekhemistri yemethylene blue; i-ion yeklorayidi enentlawulo echaseneyo ayiyo efaka umbala.

Inkqubo eqhelekileyo yokufundisa yenziwe yalula ngabom: yenza umqatyiso obhityileyo, wuyeke wome emoyeni, uwuzinzise ngobushushu, uwugqume ngemethylene blue enyibilikiswe emanzini emalunga ne-1% kangangomzuzu omnye, uwuhlambulule ngobunono ngamanzi, uwomise ngokucofa ngephepha elifunxayo, uze uwuhlolisise ngelensi entywiliselwe kwioyile. Inkqubo yokufundisa yeLibreTexts inika kanye le ndlela ye-1% yomzuzu omnye, ngoxa iphawula ukuba ezinye iilebhu ziyalandisa ixesha. Akukho sibopheleli sedayi, akukho sisusi sombala, akukho dayi yesibini. Oko kungabikho kwazo yeyona nto ibalulekileyo. Xa kukho idayi enye kuphela kwaye kungekho kususwa kwayo ngokukhetha, yonke iseli ebophelela idayi iphela inombala ofanayo.

Kwi-E. coli, indawo elindelekileyo ineeseli ezizintonga eziluhlaza (iibacilli), ezizodwa okanye ezingazimbini okanye ezikumaqela amancinane. Umbala oluhlaza uvela kwidayi. Imilo yentonga ivela kwisidalwa. Kodwa ukudityaniswa kwezi zinto akusichongi isidalwa, kuba iintlobo ezininzi zimile okweentonga yaye ezininzi zazo ziba luhlaza xa kufakwa umbala olula. I-ATCC iwucacisa lo mda wokubonisa imilo kuphela kwisikhokelo sayo semikhuliso yemicrobiology. Ingxelo echanekileyo ithi: "kubonwe iibacilli eziluhlaza." Nantoni na engaphaya kwemilo nolungelelwaniso ifuna ubungqina obungakumbi.

Ukufaka umbala ngeGram: iireyijenti ezine, uvavanyo olunye lokuziphatha

Ukufaka umbala ngeGram yindlela yokufaka umbala eyahlulayo. Ibeka zonke iiseli kulandelelwano olufanayo ize ibonise ukuba zeziphi iiseli ezigcina idayi yokuqala neziphi eziphulukana nayo. Iinkqubo zokufaka umbala ngeGram zeAmerican Society for Microbiology zichaza ulandelelwano oluqhelekileyo lwamanyathelo amane: icrystal violet (idayi yokuqala), i-iodine yeGram (isibopheleli sedayi), ialcohol okanye ialcohol eneacetone (isisusi sombala), kuze kulandele isafranin (idayi yesibini).

Imodeli eqhelekileyo yokufundisa ichaza olu lwahlulo ngesakhiwo sesigqubuthelo seseli. Iiseli zeGram-positive zinodonga olungqindilili lwepeptidoglycan oluphulukana namanzi luze lugcine indibaniselwano yecrystal violet ne-iodine ngexesha lokuhlamba ngealcohol. Iiseli zeGram-negative zinomaleko obhityileyo wepeptidoglycan kunye nenwebu yangaphandle enee-lipid ezininzi, ethi iphazanyiswe sisusisi sombala, ngoko indibaniselwano iyahlanjwa iphume zize iiseli zifunxe isafranin. Uphononongo lweGram lweNCBI Bookshelf lunika le nkcazelo yodonga lweseli. Iingcali zongeza ukuba iinkqubo ezipheleleyo zefiziksi nekhemistri zintsonkothe ngakumbi kunobungqindilili bodonga kuphela, kodwa ekutolikeni iziphumo elebhu umgaqo wokuziphatha uyasebenza: umfusa uthetha ukuba idayi igciniwe, upinki uthetha ukuba idayi ilahlekile kwaza kwafakwa idayi yesibini.

Iiphaneli zokubonisa. Yimizobo enesimbo yokuthelekisa iindlela, kungekhona iifoto zemikroskopu ezithathwe ngekhamera. Imibala yenziwe yalula ukuze kufundiswe ingqiqo: umbala omnye ekufakeni umbala olula, imibala emibini emva kokususwa kombala kwindlela yeGram.

Ingaba imethylene blue ingasetyenziswa ekufakeni umbala ngeGram

Lo mbuzo uneentsingiselo ezimbini, kwaye zifuna iimpendulo ezahlukileyo.

Intsingiselo yokuqala: ingaba ukugquma isampulu ngemethylene blue kube kanye kungathatha indawo yolandelelwano lonke lweGram. Hayi. Ngaphandle kwe-iodine, isisusi sombala, nedayi yesibini akukho nyathelo lokwahlula, ngoko akukho didi lweGram luvavanywayo. Indawo eluhlaza emva kokusebenzisa imethylene blue yodwa sisiphumo sokufaka umbala olula, nokuba ibhotile ibekwe ecaleni kweereyijenti zeGram. Musa ukuyirekhoda njengesiphumo seGram.

Intsingiselo yesibini: ingaba imethylene blue ingathatha indawo yesafranin njengedayi yesibini kwinyathelo 4. Ngokusebenza kwayo, inokufaka umbala kwiiseli ezisuswe umbala, kwaye umsebenzi okwincwadi yokufunda uqikelela iiseli zeGram-positive ezimfusa neeseli zeGram-negative eziluhlaza emva kotshintsho olunjalo. Le ngqiqo ichanekile kuba iiseli zeGram-negative ezisuswe umbala ngokuchanekileyo ziphantse zingabi nambala kwaye ziya kufunxa enye idayi yecation. Kodwa ayisiyonkqubo eqhelekileyo. Iinkqubo zeASM zichaza isafranin, kunye nezinye iidayi ezamkelweyo ezifana nebasic fuchsin kwezinye iinguqulelo, kungekhona imethylene blue. Kukwakho nengxaki ebonakalayo: kunzima ngakumbi ukwahlula uluhlaza nomfusa kunokwahlula upinki nomfusa, ngakumbi kubaqalayo naxa kusetyenziswa izibane zemikroskopu ezibuthathaka. Kumsebenzi wezifundo nakuso nasiphi na isiphumo esibalulekileyo, sebenzisa idayi yesibini echazwe yinkqubo yakho, edla ngokuba yisafranin, uze ukuthathe ukusetyenziswa kwemethylene blue njengedayi yesibini njengenguqulelo yokubonisa, kungekhona indlela eqhelekileyo elinganayo.

Umbuzo onxulumeneyo wokuba imethylene blue ithelekiseka njani nezinye iindlela zedayi enye uxoxwa kuthelekiso lweendlela zokufaka umbala olula kwiibhaktheriya.

Oko ukufaka umbala ngemethylene blue kwi-E. coli kukungqinayo, noko kungakungqiniyo

I-E. coli yibhaktheriya emile okwentonga yeGram-negative, kodwa eso sivakalisi sidibanisa iinkcukacha ezivela kwiindlela ezininzi: imilo evela kwimicroscopy, indlela esabela ngayo kwiGram evela ekufakeni umbala ngeGram, nokuchongwa kwayo okuvela kumkhuliso kunye novavanyo lwebhayokhemistri okanye lweemolekyuli. Umqatyiso wemethylene blue unegalelo kwinxalenye yokuqala kuphela. Uqinisekisa ukuba kukho iiseli ezimile okweentonga kwisampulu efakwe umbala kwaye ubonisa indlela ezilungelelene ngayo. Awuqinisekisi indlela ezisabela ngayo kwiGram, kuba akukho vavanyo lokususa umbala lwenziweyo. Awuchongi uhlobo, kuba iiseli ezizintonga zeentlobo ezininzi zikhangeleka ngokufanayo kulo mgangatho wokwahlula iinkcukacha.

Kulapho iingxelo ziphosisa khona ngendlela enokuqikelelwa. Umbhali ufaka ulwazi lwangaphambili ("lo mkhuliso ufanele ukuba yi-E. coli, kwaye i-E. coli yiGram-negative") kumgca ochaza oko kubonwayo, ukuze isilayidi sibonakale siqinisekisa into ebicingelwe nje. Ingxelo ecace ngakumbi iyazahlula ezi nxalenye: "Ukufaka umbala olula ngemethylene blue: kubonwe iibacilli eziluhlaza; yimilo kuphela, akukho didi lweGram luvavanyiweyo." Emva koko, ukuba ukufaka umbala ngeGram kwenziwe ngokuchanekileyo kusetyenziswa iisampulu zolawulo: "Ukufaka umbala ngeGram: iiseli ezizintonga ezipinki, ezihambelana neGram-negative." Ukuchongwa kohlobo kuza kamva, kwinkqubo yokuchonga, kungekhona kumbala wodwa.

Isikhokelo esisebenzisekayo sokutolika

Sebenzisa lo mgaqo wokwenza isigqibo xa usebenza elebhu. Ukuba zonke iiseli zinombala omnye kwaye inkqubo isebenzise idayi enye kungekho sisusi sombala, wenze ukufaka umbala olula. Xela imilo, uqikelelo lobungakanani, nolungelelwaniso. Yima apho. Ukuba inkqubo isebenzise icrystal violet, i-iodine, isisusi sombala, nedayi yesibini kunye neesampulu zolawulo ezaziwayo zeGram-positive neGram-negative, wenze ukufaka umbala ngeGram. Xela umbala kunye nemilo: iicocci ezimfusa ezikumatyathanga, iiseli ezizintonga ezipinki, njalo njalo.

Iimpazamo ezintathu zibangela uninzi lokubhideka. Eyokuqala, ukutolika iiseli ezizintonga eziluhlaza ngokungathi ziqinisekisa i-E. coli. Eyesibini, ukubhala isilayidi semethylene blue phantsi kwesihloko esithi "Ukufaka umbala ngeGram" kwincwadi yokubhala, nto leyo abafundi bamva abaya kuyitolika kakubi. Eyesithathu, ukutshintsha iidayi zesibini kungacingisiswanga, kuze kuthelekiswe imibala phakathi kweelebhu ngokungathi zisebenzise imibala efanayo. Impazamo nganye inesisombululo esifanayo: chaza indlela eyenziwe ngokwenene, uze ubange kuphela oko kuvavanywa yiloo ndlela.

Ukufaka umbala olula kunendawo yako kanye kuba kungqinela izinto ezimbalwa. Kuyakhawuleza, kuyazinyamezela iimpazamo ezincinane, kwaye kwanele xa umbuzo umalunga nemilo: ingaba kukho nantoni na, yaye imile njani. Xa umbuzo umalunga nokuhlela, yenza uvavanyo lokwahlula. Isilayidi esiluhlaza sikuxelela ukuba iiseli zikhangeleka njani. Isilayidi esimfusa okanye esipinki sikuxelela ukuba iindonga zazo ziziphatha njani. Akukho nasinye kwezi silayidi, sisodwa, esikuxelela uhlobo.