Ukufaka umbala okulula nge-methylene blue kuqhathaniswa nokufaka umbala ngendlela ye-Gram: lokho indlela ngayinye engakufakazela

Umfundi ulungisa isampula eligcotshwe kusilayidi elivela esikweni lamagciwane elibhalwe ukuthi E. coli, alimboze nge-methylene blue umzuzu owodwa, alihlanze, bese elibuka ngelensi esetshenziswa namafutha. Amaseli aluhlaza okwesibhakabhaka amise okwezinduku agcwele indawo ebonakalayo. Umbiko waselebhu uthi: "Amagciwane amise okwezinduku e-Gram-negative ahlonzwe njenge-E. coli." Lokho okubonakele kulungile. Kodwa leso silayidi asisekeli leso siphetho.

Iphutha lisekuqondeni umqondo, hhayi ekwenzeni inqubo. Umfundi uthathe ukufaka umbala okulula njengokungathi ukuhlolwa okuhlukanisayo. Ukufaka umbala okulula nokufaka umbala ngendlela ye-Gram kuphendula imibuzo ehlukene ngoba kusekelwe emiqondweni ehlukene yokufaka umbala. Enye ibuza ukuthi: yikuphi ukuma okukhona. Enye ibuza ukuthi: udonga lweseli luziphatha kanjani lapho luvivinywa. Uma ukudida lokhu okubili, isilayidi esifakwe umbala ngendlela efanele singaholela embikweni ongasekelwe ubufakazi.

Lokhu kuqhathanisa kugcina lezi zindlela ezimbili zihlukene, kubonise lokho umphumela ngamunye okuvumela ukuba ukusho, futhi kuphendule imibuzo emibili abafundi abayibuza kakhulu: ingabe i-methylene blue ingasetshenziswa ekufakeni umbala ngendlela ye-Gram, futhi ukufaka umbala ku-E. coli nge-methylene blue kubonisani ngempela.

Impendulo emfushane

Ukufaka umbala okulula nge-methylene blue kwenza wonke amaseli angafakwa umbala abe nombala ofanayo oluhlaza okwesibhakabhaka, ngakho kuveza ukuma kwawo (izinduku, izimbulunga, izimo ezisongene) nokuhleleka kwawo (ngalinye, ngamabili, ngamaketanga), kodwa akusho lutho ngesigaba se-Gram. Ukufaka umbala ngendlela ye-Gram kwengeza i-iodine, isinyathelo sokususa umbala, kanye nodayi wesibili, ngakho kuhlukanisa amagciwane abe yi-Gram-positive (ansomi) ne-Gram-negative (aphinki noma abomvu) ngokusekelwe endleleni izingqimba eziwazungezile ezigcina ngayo udayi. Amaseli aluhlaza okwesibhakabhaka amise okwezinduku ngemva kokusebenzisa i-methylene blue asho ukuthi "kukhona amaseli amise okwezinduku." Amaseli aphinki amise okwezinduku ngemva kokulandelana okuphelele kwezinyathelo ze-Gram asho ukuthi "kukhona amagciwane e-Gram-negative amise okwezinduku." Isitatimende sesibili sinobufakazi obunamandla kakhulu ngoba inqubo yenze ukuhlolwa okwengeziwe.

Ukuze ufunde ngeqembu elibanzi lezindlela zokufaka umbala emagciwaneni ngodayi owodwa lesi sihloko esiyingxenye yalo, bheka i-atlasi yokufaka umbala ekubukeni ngesibonakhulu.

Ukufaka umbala okulula: udayi owodwa, umbuzo owodwa

I-methylene blue ingudayi oyisisekelo (one-cation). I-ion yayo enombala inomthamo kagesi ophozithivu futhi ibophezela ezingxenyeni zeseli ezinomthamo kagesi onegethivu, kuhlanganise nama-nucleic acid namaqembu ane-asidi ebusweni beseli, ngakho amaseli abonakala eluhlaza okwesibhakabhaka phezu kwesizinda esiphaphathekile. I-OpenStax ichaza lo mgomo wokufaka umbala eselini esahlukweni sayo sokufaka umbala. I-cation kadayi uqobo iyi-ion encane eyisicaba ye-phenothiazine echazwe ngokuningiliziwe encazelweni yamakhemikhali e-methylene blue; i-ion ye-chloride ehambisana nayo ayiyona efaka umbala.

Inqubo evamile yokufundisa yenziwe yaba lula ngamabomu: gcoba ungqimba oluncane lwesampula kusilayidi, ulomise emoyeni, ulunamathisele ngokushisa, ulumboze ngesisombululo se-methylene blue esisemanzini esingaba ngu-1% cishe umzuzu owodwa, uluhlanze ngobumnene ngamanzi, ulomise ngokuluthinta ngephepha elimuncayo, bese uluhlola ngelensi esetshenziswa namafutha. Inqubo yokufundisa ye-LibreTexts inikeza yona kanye le ndlela ka-1% yomzuzu owodwa, iphinde iphawule ukuthi amanye amalebhu andisa isikhathi. Asikho isibophezeli sikadayi, akukho okususa umbala, akukho dayi wesibili. Yilokho kanye okubalulekile. Ngodayi owodwa kuphela futhi kungabikho ukususwa okukhethiwe, wonke amaseli abophezela udayi agcina enombala ofanayo.

Ku-E. coli, okulindeleke ukuba kubonakale ngamaseli aluhlaza okwesibhakabhaka amise okwezinduku (ama-bacilli), ngalinye noma ngamabili noma ngamaqoqo amancane. Umbala oluhlaza okwesibhakabhaka uvela kudayi. Ukuma njengenduku kuvela egciwaneni. Kodwa le nhlanganisela ayiluhlonzi uhlobo lwegciwane, ngoba izinhlobo eziningi zimise okwezinduku futhi eziningi zazo ziba luhlaza okwesibhakabhaka lapho zifakwa umbala okulula. I-ATCC ikubeka ngokucacile lokhu ukuthi umphumela ukhawulelwe ekumeni kuphela emhlahlandleleni wayo wokukhulisa amagciwane. Umbiko ofanele uthi: "kubonakale ama-bacilli aluhlaza okwesibhakabhaka." Noma yini engaphezu kokuma nokuhleleka idinga ubufakazi obengeziwe.

Ukufaka umbala ngendlela ye-Gram: izithako ezine, ukuhlolwa okukodwa kokuziphatha

Ukufaka umbala ngendlela ye-Gram kuyindlela yokufaka umbala ehlukanisayo. Kudlulisa wonke amaseli ekulandelaneni okufanayo kwezinyathelo bese kubhekwa ukuthi yimaphi agcina udayi wokuqala nokuthi yimaphi awulahlekelwayo. Izinqubo zokufaka umbala ngendlela ye-Gram ze-American Society for Microbiology zichaza ukuhleleka okujwayelekile kwezinyathelo ezine: i-crystal violet (udayi wokuqala), i-iodine ye-Gram (isibophezeli sikadayi), utshwala noma ingxube ye-acetone notshwala (okususa umbala), bese kuba yi-safranin (udayi wesibili).

Incazelo evamile yokufundisa ichaza lokhu kuhlukana ngokwakheka kwezingqimba ezizungeze iseli. Amaseli e-Gram-positive anodonga oluwugqinsi lwe-peptidoglycan oluphelelwa amanzi bese lugcina inhlanganisela ye-crystal violet ne-iodine ngesikhathi kuhlanzwa ngotshwala. Amaseli e-Gram-negative anongqimba oluncane lwe-peptidoglycan kanye nolwelwesi lwangaphandle olunamafutha amaningi oluphazanyiswa yilokho okususa umbala, ngakho inhlanganisela iyagezeka iphume bese amaseli emunca i-safranin. Ukubuyekezwa kokufaka umbala ngendlela ye-Gram ku-NCBI Bookshelf kunikeza le ncazelo yodonga lweseli. Ochwepheshe bengeza ukuthi zonke izinqubo zomzimba nezamakhemikhali ezihilelekile ziyinkimbinkimbi kakhulu kunobukhulu bodonga kuphela, kodwa ekuchazeni imiphumela elebhu umthetho wokuziphatha usasebenza: onsomi usho ukuthi udayi ugciniwe, ophinki usho ukuthi udayi ulahlekile kwase kufakwa owesibili.

Imidwebo yokubonisa. Imidwebo eyenziwe ngesitayela ukuze kuqhathaniswe izindlela, hhayi izithombe ezithathwe ngekhamera yesibonakhulu. Imibala yenziwe yaba lula ukuze kufundiswe umqondo: umbala owodwa ekufakeni umbala okulula, imibala emibili ngemva kokususa umbala ngendlela ye-Gram.

Ingabe i-methylene blue ingasetshenziswa ekufakeni umbala ngendlela ye-Gram

Lo mbuzo unezincazelo ezimbili, futhi zidinga izimpendulo ezihlukene.

Incazelo yokuqala: ingabe ukumboza isampula nge-methylene blue kanye kuphela kungathatha indawo yakho konke ukulandelana kwezinyathelo ze-Gram. Cha. Ngaphandle kwe-iodine, okususa umbala, kanye nodayi wesibili, asikho isinyathelo esihlukanisayo, ngakho asikho isigaba se-Gram esihlolwayo. Indawo eluhlaza okwesibhakabhaka ngemva kokusebenzisa i-methylene blue kuphela ingumphumela wokufaka umbala okulula ngisho noma ibhodlela libekwe eduze kwezithako ze-Gram. Ungawubhali njengomphumela we-Gram.

Incazelo yesibili: ingabe i-methylene blue ingathatha indawo ye-safranin njengodayi wesibili esinyathelweni sesi-4. Ngokusebenza ingafaka umbala kumaseli asuswe umbala, futhi umsebenzi osencwadini yokufunda ubikezela amaseli e-Gram-positive ansomi namaseli e-Gram-negative aluhlaza okwesibhakabhaka ngemva kwalokho kushintsha. Lo mqondo unengqondo ngoba amaseli e-Gram-negative asuswe umbala kahle acishe angabi nambala futhi azomunca omunye udayi one-cation. Kodwa lena akuyona inqubo ejwayelekile. Izinqubo ze-ASM zicacisa i-safranin, zinezinye izithako ezaziwayo njenge-basic fuchsin kwezinye izinhlobo zenqubo, hhayi i-methylene blue. Kukhona nenkinga ekusebenzeni: kunzima kakhulu ukuhlukanisa oluhlaza okwesibhakabhaka nonsomi kunokuhlukanisa ophinki nonsomi, ikakhulukazi kwabaqalayo nangaphansi kwezibani zesibonakhulu ezingasakhanyi kahle. Emisebenzini yokufunda nakunoma yimuphi umphumela obalulekile, sebenzisa udayi wesibili oshiwo yinqubo yakho, ngokuvamile i-safranin, futhi uthathe ukusetshenziswa kwe-methylene blue njengodayi wesibili njengendlela ehlukile yokubonisa kunokuba kube indinganiso elinganayo.

Umbuzo ohlobene wokuthi i-methylene blue iqhathaniswa kanjani nezinye izindlela zikadayi owodwa uchazwe ekuqhathanisweni kwezindlela zokufaka umbala okulula emagciwaneni.

Lokho ukufaka umbala ku-E. coli nge-methylene blue okukufakazela, nalokho okungakufakazeli

I-E. coli iyigciwane le-Gram-negative elimise okwenduku, kodwa lowo musho uhlanganisa amaqiniso avela ezindleleni eziningana: ukuma kutholakala ngokubuka ngesibonakhulu, ukusabela kwe-Gram ngokufaka umbala ngendlela ye-Gram, ukuhlonza uhlobo ngokukhulisa amagciwane kanye nokuhlolwa kwamakhemikhali emvelo noma kwama-molecule. Isampula eligcotshwe kusilayidi lafakwa i-methylene blue linikela ngengxenye yokuqala kuphela. Lifakazela ukuthi amaseli amise okwezinduku akhona esampuleni esifakwe umbala nokuthi ahleleke kanjani. Alifakazeli ukusabela kwe-Gram, ngoba akukho kuhlolwa kokususa umbala okwenziwe. Aliluhlonzi uhlobo lwegciwane, ngoba amagciwane amise okwezinduku ezinhlobo eziningi abukeka efana kuleli zinga lokuhlukanisa imininingwane.

Yilapho imibiko iphambuka khona ngendlela elindelekile. Umbhali ufaka ulwazi lwangaphambili ("leli siko lamagciwane kufanele libe yi-E. coli, futhi i-E. coli iyi-Gram-negative") emgqeni walokho okubonakele, ngakho isilayidi sibonakala siqinisekisa lokho obekucatshangwa nje. Umbiko ocace kakhudlwana uhlukanisa lezi zingxenye: "Ukufaka umbala okulula nge-methylene blue: kubonakale ama-bacilli aluhlaza okwesibhakabhaka; ukuma kuphela, asikho isigaba se-Gram esihloliwe." Bese, uma ukufaka umbala ngendlela ye-Gram kwenziwe kahle kusetshenziswa amasampula okulawula: "Ukufaka umbala ngendlela ye-Gram: amaseli aphinki amise okwezinduku, ahambisana ne-Gram-negative." Ukuhlonzwa kohlobo lwegciwane kuza kamuva, ngenqubo yokuhlonza, hhayi ngombala kuphela.

Umhlahlandlela osebenzisekayo wokuchaza imiphumela

Sebenzisa lo mthetho wokunquma lapho usebenza elebhu. Uma wonke amaseli enombala owodwa futhi inqubo isebenzise udayi owodwa ngaphandle kokususa umbala, wenze ukufaka umbala okulula. Bika ukuma, isilinganiso sobukhulu, nokuhleleka. Yima lapho. Uma inqubo isebenzise i-crystal violet, i-iodine, okususa umbala, kanye nodayi wesibili, kanye namasampula okulawula aziwayo e-Gram-positive ne-Gram-negative, wenze ukufaka umbala ngendlela ye-Gram. Bika umbala kanye nokuma: ama-cocci ansomi ngamaketanga, amaseli aphinki amise okwezinduku, nokunye.

Amaphutha amathathu abangela ukudideka okuningi. Elokuqala, ukuthatha amaseli aluhlaza okwesibhakabhaka amise okwezinduku njengokuqinisekiswa kwe-E. coli. Elesibili, ukubhala isilayidi se-methylene blue ngaphansi kwesihloko esithi "Ukufaka umbala ngendlela ye-Gram" encwadini yamanothi, okuyokwenza abafundi bakamuva basichaze ngokungafanele. Elesithathu, ukushintsha odayi besibili ngokunganaki bese kuqhathaniswa imibala phakathi kwamalebhu njengokungathi amasethi emibala ayafana. Iphutha ngalinye linesixazululo esifanayo: yisho indlela eyenziwe ngempela, futhi usho kuphela lokho leyo ndlela ekuhlolayo.

Ukufaka umbala okulula kubalulekile ngoba kusho okuncane kuphela. Kuyashesha, kuyakwazi ukubekezelela ukuphambuka okuncane enqubweni, futhi kwanele lapho umbuzo umayelana nokuma: ingabe kukhona okukhona, futhi kumise kanjani. Lapho umbuzo umayelana nokubeka ezigabeni, yenza ukuhlolwa okuhlukanisayo. Isilayidi esiluhlaza okwesibhakabhaka sikutshela ukuthi amaseli abukeka kanjani. Isilayidi esinsomi noma esiphinki sikutshela ukuthi izindonga zawo ziziphatha kanjani. Asikho kulezi zilayidi, sisodwa, esikutshela uhlobo lwegciwane.