Kuchazwa odayi be-methylene blue kaLoeffler abane-alkali nabe-polychrome

Amabhodlela amathathu angahlala eduze kwelinye ebhentshini lokudaya, ngalinye linelebula lohlobo oluthile lwe-methylene blue. Elinye lithi i-methylene blue esemanzini engu-1%. Elinye lithi i-methylene blue kaLoeffler ene-alkali. Elinye lithi i-polychrome methylene blue. Isisebenzi esimatasa sithatha eliseduze kakhulu. Isampula eligcotshwe ku-slide libe selidayeka ngokuphaphathekile kakhulu, noma libe luhlaza okwesibhakabhaka ngokufanayo yonke indawo, noma libe nemithunzi ensomi engalindelekile, bese umphumela usolwa kusampula.

Lena inkinga yokwakheka kwesixazululo, hhayi eyesampula. Lezi zixazululo ezintathu ziqukethe odayi abahlobene kodwa zimelela imiqondo ehlukene yokudaya. Esinye siwudayi olula one-charge ephozithivu ku-pH eseduze nokungathathi hlangothi. Esinye siwudayi ofanayo owenziwe wasebenza ngamandla engeziwe nge-alkali. Esinye asisesiwudayi owodwa nhlobo, kodwa sesiyingxube eyenziwe i-oxidation ngamabomu. Uma usebenzisa esingafanele, umgomo wokudaya uyashintsha lapho usebenza.

Impendulo emfushane

I-methylene blue ejwayelekile esemanzini iyisisekelo: cishe udayi ongu-1% emanzini, udayi olula one-charge ephozithivu wokuhlola ukwakheka kwamabhaktheriya nokuhlola okuvamile ngesibonakhulu. I-methylene blue kaLoeffler ene-alkali yengeza inani elincane elilinganisiwe le-alkali ehlanjululiwe esixazululweni esiyisisekelo esinotshwala esihlanjululwe ngamanzi; lokhu kukhuphula i-pH futhi kwenze udayi one-charge ephozithivu unamathele ngamandla ezakhiweni zamabhaktheriya ezine-asidi. I-polychrome methylene blue iyi-methylene blue evuthiswe ngamabomu nge-oxidation ku-alkali, ngakho ibhodlela liqukethe ingxube yama-homologue asuswe amaqembu e-methyl (azure A, azure B, azure C), i-thionin, ne-methylene violet kanye ne-methylene blue esele; le ngxube idaya izakhiwo ezihlukene ngemibala ehlukene.

Uma ukhumbula umusho owodwa kuphela: ukufaka i-alkali kushintsha amandla okudaya, kanti ukwenza i-polychrome kushintsha imibala yokudaya.

I-methylene blue ejwayelekile esemanzini: isisekelo sokuqhathanisa

Ukwakheka okulula kakhulu yi-methylene blue encibilikiswe emanzini, ngokuvamile engu-1% w/v, ngaphandle kwe-alkali nangaphandle kwe-oxidation. Ingxenye enombala iyi-cation ye-methylthioninium ene-ion ye-chloride eneshaji ephambene, udayi omncane oyisicaba we-phenothiazine ochazwa embhalweni we-PubChem nge-methylene blue ngokuthi yi-methylthioninium chloride. Njenge-cation, ibopha uchungechunge lwe-phosphate olusekela ama-nucleic acid kanye namaqembu ane-asidi asezindongeni zamaseli amabhaktheriya, idaye amaseli abe luhlaza okwesibhakabhaka ngemuva elikhanyayo.

Okuhle ngayo ukuthi imiphumela yayo iyabikezeleka. Ama-nucleus namabhaktheriya abonakala eluhlaza okwesibhakabhaka. Ukugqama komahluko kuphakathi. Izinto eziphilayo ezingabonakali kahle zingadayeka ngokubuthakathaka, okuyilowo mkhawulo uLoeffler ayehlose ukuwulungisa. Ukuze wazi kabanzi ngomndeni obanzi wodayi abalula bamabhaktheriya lesi sixazululo esiyingxenye yawo, bheka uhlolojikelele lokudaya amabhaktheriya ngodayi olula. Ukuqinisekisa ukuthi into yiyona ngempela kusabalulekile lapha: i-methyl blue iyi-molecule ehluke ngokuphelele, njengoba kuchazwa ku-methyl blue uma iqhathaniswa ne-methylene blue, ngakho ukusesha ikhathalogi usebenzisa igama lombala kuphela kungakunika isigaba sikadayi esingafanele.

I-methylene blue kaLoeffler ene-alkali: udayi ofanayo, osebenza ngamandla engeziwe

Indlela yakudala kaLoeffler ihlanganisa isixazululo esiyisisekelo se-methylene blue esinotshwala ne-alkali ehlanjululiwe emanzini. Izinguqulo ezisezincwadini zokufunda zidinga udayi ongu-0.3 g ku-30 mL we-ethanol engu-95%, kuhlanganiswe no-100 mL we-potassium hydroxide engu-0.01% esemanzini (noma, ngokulinganayo, u-30 mL we-methylene blue ogcwele esixazululweni sotshwala, u-100 mL wamanzi acwengwe nge-distillation, no-1 mL we-KOH engu-1%), okwenza isixazululo esisetshenziswayo sibe nodayi ocishe ube ngu-0.3% ku-pH ene-alkali ngokusobala. Izinguqulo ezithengiswayo zilandela umgomo ofanayo, futhi ikhathalogi kaThermo Fisher yodayi besayensi yezinto eziphilayo ezincane ibala i-methylene blue kaLoeffler ene-alkali kanye nezinhlobo ze-polychrome nezisemanzini njengemikhiqizo ehlukene yekhathalogi enezinjongo ezihlukene zokusetshenziswa.

I-alkali iyona eyenza umehluko. Ukukhuphula i-pH kususa ama-proton emaqenjini e-carboxyl nawe-phosphate akusampula, kwandise ukuminyana kwezindawo ezine-charge enegethivu, kuyilapho isixazululo esiyisisekelo se-ethanol sigcina udayi uncibilike ngokuphelele ukuze ufike kulezo zindawo usesimweni sama-monomer. Umphumela uba ukudayeka kwamabhaktheriya ngokushesha nangamandla engeziwe kunokudayeka okuvela kudayi osemanzini ongathathi hlangothi, kube nezinduku nama-cocci aluhlaza okwesibhakabhaka ngokujulile ajwayelekile kumasampula amabhaktheriya agcotshwe kuma-slide. Isikhathi sokudaya ngokuvamile siwumzuzu owodwa kuya kwemithathu, kulandelwe ukugeza ngamanzi.

Ukusetshenziswa kwakudala kuse-Corynebacterium diphtheriae ekhuliswe esisekelweni sokukhulisa se-serum sikaLoeffler: izinduku ezincane, ezivame ukuvuvukala emaphethelweni, ezinama-granule e-Babes-Ernst ashintsha umbala lapho edayiwe futhi adayeka ngamandla, aqukethe i-polyphosphate egciniwe. Lokhu kuhlanganiswa kwesisekelo sokukhulisa kanye nodayi kunomlando, kokubili kuhlonishwa usosayensi ofanayo wamabhaktheriya, futhi ukugqama kwama-granule kuyinzuzo ekuxilongeni. Ngaphandle komsebenzi we-diphtheria, lo dayi usebenza njengodayi ojwayelekile wamabhaktheriya kumasampula agcotshwe kuma-slide avela ezikhulisweni, ukuhlola ukwakheka kwe-Neisseria kumasampula omtholampilo agcotshwe kuma-slide, nanoma kuphi lapho ukudaya nge-Gram kungadingekile kodwa udayi oluhlaza okwesibhakabhaka ojwayelekile osemanzini uphaphatheka kakhulu. Ungudayi wombala owodwa. Konke okune-asidi kuba nomthunzi othile woluhlaza okwesibhakabhaka. Uma inqubo idinga imibala ehlukanisayo, i-alkali iyodwa ayikwazi ukuyinikeza.

Isixwayiso esisodwa esisebenzayo: izixazululo zikadayi ezine-alkali ziyawohloka futhi zakhe inzika lapho zigcinwa futhi zimunca i-carbon dioxide. Amalabhorethri alungisa amanani angemaningi noma athenge isixazululo sekhathalogi futhi ahloniphe isikhathi saso sokusebenziseka, kunokulokhu esebenzisa ibhodlela elidala eselifaka inzika kuyo yonke i-slide.

I-polychrome methylene blue: igama elilodwa, odayi abaningana

I-polychrome methylene blue iqala ngodayi ofanayo kodwa iwufaka ku-oxidation elawulwayo ku-alkali, okwakubizwa ngokomlando ngokuthi ukuvuthisa noma ukwenza i-polychrome. Ukuguga ku-alkali, ukushisisa nge-carbonate, noma i-oxidation ngezinto ezifana ne-dichromate kususa amaqembu e-methyl ezingxenyeni ze-dimethylamino, igxathu negxathu. Umkhiqizo uba yingxube: i-methylene blue esele kanye ne-azure B (kususwe i-methyl eyodwa), i-azure A ne-azure C (kususwe amaqembu e-methyl engeziwe), i-thionin (kususwe wonke amaqembu e-methyl), kanye nemikhiqizo eseceleni ye-oxidation yomndeni we-methylene violet. Ingxenye ngayinye inombala wayo namandla ayo okunamathela ezicutshini, ngakho ingxube idaya ngokushintsha umbala: ama-nucleus, ama-granule, ama-mucin, noqwanga ngakunye kuthatha imithunzi ehlukene esukela koluhlaza okwesibhakabhaka kuya kobomvu-nsomi.

Le khemistri iyona futhi eyisisekelo sodayi begazi abadumile. Abacwaningi bakuqala bathola ukuthi i-methylene blue efakwe i-alkali yayidaya ama-nucleus ezimuncagazi zikamalaleveva ngemithunzi ensomi, kodwa izixazululo zabo ezivuthisiwe ezisemanzini zazingazinzile futhi zakha inzika ezinsukwini ezimbalwa. Ukubuyekezwa kokuthi udayi wegazi onsomi wasungulwa kanjani kulandisa ukuthi uMalachowski noRomanowsky bawusebenzisa kanjani lo mphumela ngokuzimela ngo-1891, ukuthi i-methanol yethulwa kanjani ukuze kuzinziswe inzika kadayi futhi kuqiniswe izingqimba zesampula ngaphandle kokushisa, nokuthi kamuva uGiemsa wenza kanjani ingxube yaba nesilinganiso esifanayo esisekelwe ku-azure B elinganisiwe kanye ne-eosin, izinzi­swe nge-glycerol. Odayi banamuhla bohlobo lwe-Giemsa bavela ngokuqondile ku-polychrome methylene blue kanye ne-eosin, inhlanganisela eyenza umphumela we-Romanowsky. Ukuze uthole imininingwane ngokuhlanganiswa ne-eosin nangengxenye ye-azure B, bheka odayi bohlobo lwe-Romanowsky nokusebenzisana kwe-azure B ne-eosin.

Ngenxa yokuthi iyingxube, i-polychrome methylene blue ngokwemvelo iyona enokulinganiselwa okungafani kakhulu kulezi ezintathu. Ukulingana kwayo okuqondile kwemibala kuncike endleleni nasezingeni le-oxidation, yingakho abahlinzeki bemikhiqizo yokuhlola izicubu beyithengisa njengomkhiqizo ohlukile onokuhlolwa kokusebenza kwawo, hhayi njengekaLoeffler esigugile. Ukusetshenziswa okuvamile kuhlanganisa ukubonisa ama-granule amaseli e-mast, ama-mucin noqwanga ngokushintsha umbala lapho kudayiwa, imininingwane yamaseli e-plasma, kanye ne-Helicobacter pylori ezincezwini zezicubu ngezindlela ze-polychrome zohlobo lwe-Goodpasture. Iyona efanele uma umbuzo uwukuhlukanisa ngombala, futhi ayifanele uma inqubo ilindele umumo webhaktheriya ocacile onombala owodwa.

Ukuqhathanisa ukwakheka nenhloso

Funda ithebula njengendlela yokwenza isinqumo. Uma isampula lidayeka ngokuphaphathekile kakhulu, shintsha umqondo we-pH bese usebenzisa ekaLoeffler. Uma konke kudayeka kodwa kungabi khona okuhlukanisekayo, shintsha umqondo kadayi bese usebenzisa eye-polychrome. Uma isampula eligcotshwe ku-slide ladaywa ngekaLoeffler libonisa ama-granule kahle kakhulu kodwa umphathi ebuza ukuthi kungani ama-nucleus ne-cytoplasm kunombala ofanayo oluhlaza okwesibhakabhaka, impendulo iwukuthi awukho udayi wombala owodwa one-alkali ongawenza lowo msebenzi. Lowo mbuzo ungowomndeni we-polychrome kanye nezinhlobo zodayi begazi ezivela kuwo.

Ukugwema ukudideka okuvamile

Amaphutha amabili adala ukudideka okuningi. Elokuqala ukuthatha ekaLoeffler njengengashintshaniswa ne-methylene blue ejwayelekile ngoba yomibili ibukeka iluhlaza okwesibhakabhaka ebhodleleni. Yehlukile nge-pH nangamandla okudaya, futhi ukufaka udayi ongathathi hlangothi lapho kucaciswe ekaLoeffler kulahlekelisa yona kanye indlela yokugqamisa ama-granule inqubo eyakhelwe kuyo. Elesibili ukuthatha noma yimuphi udayi omdala oluhlaza okwesibhakabhaka one-alkali, noma osugugile ngephutha, njenge-polychrome. Ukuguga okungalawulwa kukhiqiza ingxube engaziwa ngamandla angaziwa, kuyilapho izixazululo zangempela ze-polychrome zilungiswa ngezindlela ezichaziwe ze-oxidation futhi, emndenini wodayi begazi, zihlanganiswa ne-eosin elinganisiwe. Umlando ufundisa okuthile lapha: izixazululo ezivuthisiwe ezisemanzini ezazingazinzile zeminyaka yawo-1890 zathathelwa indawo yizixazululo ezizinzisiwe ezine-methanol ngenxa yokuthi izingxube ezingalawulwa zazingenakuphindwa ngokufanayo, okuyisifundo okufanele sikhunjulwe ngaphambi kokudaya i-slide yokuxilonga ngodayi osebhodleleni elingaziwa ukuthi selinesikhathi esingakanani.

Ukuqikelela amagama kuphelelisa lokhu. Qinisekisa umkhiqizo wekhathalogi (osemanzini uma uqhathaniswa nekaLoeffler noma eye-polychrome), okuqukethwe kukadayi, nokusetshenziswa okuhlosiwe kulebula, kunokukuqagela ngombala. Lezi zixazululo ezintathu zinomsuka ofanayo wengxenye enikeza umbala. Azinayo inhloso efanayo.