Ingcaciso yeedayi zikaLoeffler zemethylene blue enealkali nepolychrome

Iibhotile ezintathu zinokuma ecaleni kwenye kwibhentshi yokufaka umbala, nganye ineleyibhile yohlobo oluthile lwemethylene blue. Enye ithi yimethylene blue enyibilikiswe emanzini eyi-1%. Enye ithi yimethylene blue kaLoeffler enealkali. Enye ithi yimethylene blue yepolychrome. Umsebenzi oxakekileyo uthatha eyona ikufutshane. Emva koko isampulu eqatywe kwisilayidi ifumana umbala ongabonakali kakuhle, okanye oluhlaza okwesibhakabhaka ngokufanayo kuyo yonke indawo, okanye izithunzi ezimfusa ebezingalindelekanga, kuze kugxekwe isampulu ngenxa yesiphumo.

Le yingxaki yomxube, hayi eyesampulu. Ezi zisombululo zithathu zineedayi ezinxulumeneyo kodwa zisebenzisa imigaqo eyahlukileyo yokufaka umbala. Esinye yidayi eqhelekileyo eneeyoni ezinentshaji entle kwi-pH ekufutshane nephakathi. Esinye yidayi efanayo eyenziwe yasebenza ngamandla ngakumbi ngealkali. Esinye asiseyodayi enye kwaphela, kodwa ngumxube owenziwe ngeoksideshini ngabom. Ukuba usebenzisa engachanekanga, umgaqo wokufakwa kombala uyatshintsha ngoxa usebenza.

Impendulo emfutshane

Imethylene blue eqhelekileyo enyibilikiswe emanzini sisiseko: malunga ne-1% yedayi emanzini, idayi elula eneeyoni ezinentshaji entle esetyenziselwa ukujonga ukumila kweebhaktiriya nokusebenza ngokubanzi ngemikroskopu. Imethylene blue kaLoeffler enealkali yenziwa ngokongeza isixa esincinci esilinganisiweyo sealkali exutywe yancitshiswa kugxininiso lwedayi enyibilikiswe etywaleni lwaze lwaxutywa namanzi; oku kunyusa i-pH kuze kwenze idayi eneeyoni ezinentshaji entle ibambelele ngamandla ngakumbi kwizakhiwo zeebhaktiriya ezineasidi. Imethylene blue yepolychrome yimethylene blue evuthiswe ngabom ngeoksideshini kwialkali, ngoko ibhotile inomxube weehomologi ezisuswe amaqela emethyl (azure A, azure B, azure C), ithionin, nemethylene violet kunye nemethylene blue eseleyo, yaye loo mxube unika izakhiwo ezahlukeneyo imibala eyahlukileyo.

Ukuba ukhumbula isivakalisi esinye kuphela: ukongeza ialkali kutshintsha ubunzulu bombala, ngoxa ukwenza ipolychrome kutshintsha imibala evelayo.

Imethylene blue eqhelekileyo enyibilikiswe emanzini: isiseko sokuthelekisa

Owona mxube ulula yimethylene blue enyibilikiswe emanzini, edla ngokuba yi-1% w/v, engenaalkali nengenzwanga ngeoksideshini. Into enombala yikationi yemethylthioninium ehamba neyoni yeklorayidi enentshaji echaseneyo, idayi encinci nesicaba yephenothiazine echazwa kwirekhodi yePubChem yemethylene blue njengemethylthioninium chloride. Njengekationi, ibophelela kwimixokelelwane yephosphate yeeasidi zenukliyiki nakumaqela aneasidi asezindongeni zeeseli zeebhaktiriya, inike iiseli umbala oluhlaza okwesibhakabhaka ngokuchasene nemvelaphi ekhanyayo.

Into eluncedo ngayo kukuba iziphumo zayo ziqikeleleka. Iinucleus neebhaktiriya zibonakala ziluhlaza okwesibhakabhaka. Umahluko phakathi kwemibala awumkhulu. Izinto eziphilayo ezingabonakali kakuhle zisenokufumana umbala obuthathaka, nto leyo kanye uLoeffler awayezama ukuyilungisa. Ukuze ufumane ulwazi ngosapho olubanzi lweedayi ezilula zeebhaktiriya esi sisombululo esikulo, jonga isishwankathelo sokufakwa kombala kwiibhaktiriya ngedayi enye. Ukuqinisekisa ukuba idayi ichongwe ngokuchanekileyo kusabalulekile apha: imethyl blue yimolekyuli eyahluke ngokupheleleyo, njengoko kuchaziwe kumahluko phakathi kwemethyl blue nemethylene blue, ngoko ukukhangela kwikhathalogu usebenzisa igama lombala kuphela kunokukukhokelela kuhlobo olungachanekanga lwedayi.

Imethylene blue kaLoeffler enealkali: idayi efanayo, esebenza ngamandla ngakumbi

Iresiphi yemveli kaLoeffler idibanisa ugxininiso lwemethylene blue enyibilikiswe etywaleni nealkali exutywe yancitshiswa emanzini. Iinguqulelo ezisezincwadini zokufunda zifuna i-0.3 g yedayi kwi-30 mL ye-95% ethanol kunye ne-100 mL ye-0.01% potassium hydroxide enyibilikiswe emanzini (okanye, ngokulinganayo, i-30 mL yesisombululo esizeleyo semethylene blue etywaleni, i-100 mL yamanzi adistiliweyo, ne-1 mL ye-1% KOH), nto leyo enika isisombululo esisetyenziswayo esinedayi emalunga ne-0.3% kunye ne-pH enealkali ngokucacileyo. Iinguqulelo zorhwebo zilandela umgaqo ofanayo, yaye ikhathalogu yeThermo Fisher yeedayi zemikrobhayoloji idwelisa imethylene blue kaLoeffler enealkali kunye neenguqulelo zepolychrome nezasemanzini njengeemveliso ezahlukeneyo zekhathalogu ezineenjongo zokusetyenziswa ezahlukeneyo.

Ialkali yeyona nto iphambili esebenzayo. Ukunyusa i-pH kususa iiprotoni kumaqela ekharboksili nawephosphate akwisampulu, kwandise ukuxinana kweendawo ezinentshaji embi, ngoxa ugxininiso lwetywala lugcina idayi inyibilike ngokupheleleyo ukuze ifike kwezo ndawo ikwimo yeemonomeri. Isiphumo kukufakwa kombala kwiibhaktiriya ngokukhawuleza nangamandla ngakumbi kunoko kwenziwa yidayi esemanzini ene-pH ephakathi, kuvele iibhaktiriya ezimile okweentonga nee-cocci eziqhelekileyo ezinombala oluhlaza okwesibhakabhaka onzulu kwisampulu eqatywe kwisilayidi sokuhlola iibhaktiriya. Ixesha lokufaka umbala lidla ngokuba ngumzuzu omnye ukuya kwemithathu, kulandelwe kukuhlamba ngamanzi.

Ukusetyenziswa kwayo kwemveli kukwiCorynebacterium diphtheriae ekhuliswe kwisixhobo sokukhulisa iibhaktiriya seserum kaLoeffler: iintonga ezibhityileyo, ezidla ngokuba neziphelo ezityebileyo, ezineegranuli zeBabes-Ernst ezitshintsha umbala ngokohlobo lwesakhiwo nezifumana umbala onamandla, eziqulethe ipolyphosphate egciniweyo. Ukudityaniswa kwesixhobo sokukhulisa iibhaktiriya nedayi kunembali ende, zombini zinxulunyaniswa nengcali enye yeebhaktiriya, yaye umahluko obonakalayo weegranuli yeyona nzuzo ekuchongeni. Ngaphandle komsebenzi wediphtheria, le dayi isetyenziswa njengedayi eqhelekileyo yokuhlola iibhaktiriya kwiisampulu ezithathwe kwiinkcubeko zeebhaktiriya zaza zaqatyiwa kwizilayidi, ekujongeni ukumila kweNeisseria kwiisampulu zeklinikhi eziqatyiweyo, nakwiimeko apho ukufakwa kombala ngendlela kaGram kungafuneki kodwa imethylene blue eqhelekileyo esemanzini inika umbala obuthathaka kakhulu. Yidayi yombala omnye. Yonke into eneasidi ifumana isithunzi esithile sombala oluhlaza okwesibhakabhaka. Xa inkqubo ifuna imibala eyahlulayo, ialkali iyodwa ayinakuyinika.

Isilumkiso esinye esisebenzayo: izisombululo zedayi enealkali ziyawohloka zize zenze intlenga xa zigcinwa naxa zifunxa ikharbon dayoksayidi. Iilebhu zilungiselela izixa ezincinci okanye zithenge isisombululo esikwikhathalogu zize zihloniphe ixesha laso lokugcinwa, kunokuba ziqhubeke zisebenzisa ibhotile endala eshiya intlenga kwisilayidi ngasinye.

Imethylene blue yepolychrome: igama elinye, iidayi ezininzi

Imethylene blue yepolychrome iqala ngedayi efanayo kodwa yenziwa ngeoksideshini elawulwayo kwialkali, nto leyo eyayibizwa ngokuba kukuvuthiswa okanye ukwenza ipolychrome. Ukugcinwa ixesha elide kwialkali, ukufudumezwa ngecarbonate, okanye ioksideshini ngezinto ezifana nedichromate kususa amaqela emethyl kumaqela edimethylamino ngokwamanqanaba. Imveliso ngumxube: imethylene blue eseleyo kunye ne-azure B (kususwe imethyl enye), i-azure A ne-azure C (kususwe amaqela emethyl angakumbi), ithionin (kususwe onke amaqela emethyl), neemveliso ezisecaleni zeoksideshini zosapho lwemethylene violet. Icandelo ngalinye linombala walo notyekelo lwalo lokubophelela kwizicubu, ngoko umxube ufaka imibala etshintshayo ngokohlobo lwesakhiwo: iinucleus, iigranuli, iimucin, nentlala nganye ifumana izithunzi ezahlukileyo ukusuka kuluhlaza okwesibhakabhaka ukuya kobomvu omfusa.

Le khemistri ikwayile isisiseko seedayi zegazi ezidumileyo. Abaphandi bokuqala bafumanisa ukuba imethylene blue engezwe ialkali yayinika iinucleus zesifunxi-gazi semalariya izithunzi ezimfusa, kodwa izisombululo zabo ezivuthisiweyo zasemanzini zazingazinzanga yaye zazisenza intlenga kwisithuba seentsuku. Uphononongo lwendlela eyayilwa ngayo idayi yegazi emfusa luchaza indlela uMalachowski noRomanowsky abawusebenzisa ngayo lo mphumo ngokuzimeleyo ngo-1891, indlela imethanol eyaziswa ngayo ukuzinzisa intlenga yedayi nokumisela iisampulu ezicekethekileyo kwizilayidi ngaphandle kobushushu, nendlela uGiemsa awathi kamva wamisela umxube one-azure B elinganisiweyo kunye ne-eosin, uzinziswe ngeglycerol. Iidayi zanamhlanje zohlobo lweGiemsa zivela ngqo kwimethylene blue yepolychrome kunye ne-eosin, umdibaniso ovelisa umphumo weRomanowsky. Ukuze ufumane iinkcukacha zokudityaniswa ne-eosin kunye necandelo le-azure B, jonga iidayi zohlobo lweRomanowsky nentsebenziswano ye-azure B ne-eosin.

Ngenxa yokuba ingumxube, imethylene blue yepolychrome ngokwendalo yeyona incinci ekumiseleni ukufana kwayo kwezi zintathu. Ibhalansi echanekileyo yemibala yayo ixhomekeke kwindlela nakubungakanani beoksideshini, yiyo loo nto abathengisi bezinto zehistoloji beyithengisa njengemveliso eyahlukileyo enovavanyo lwayo lokusebenza, kungekhona njengemethylene blue kaLoeffler egcinwe ixesha elide. Ukusetyenziswa okuqhelekileyo kuquka ukubonisa iigranuli zeeseli zemast, iimucin, nentlala ngemibala etshintshayo ngokohlobo lwesakhiwo, ukubonisa iinkcukacha zeeseli zeplasma, nokubonisa iHelicobacter pylori kumacandelo ezicubu ngeendlela zepolychrome zohlobo lweGoodpasture. Lukhetho olufanelekileyo xa injongo ikukwahlula izinto ngombala, kodwa lukhetho olungafanelekanga xa inkqubo ilindele umfanekiso ocacileyo weebhaktiriya onombala omnye.

Ukuthelekiswa kwemixube neenjongo

Funda itheyibhile njengenkqubo yokwenza isigqibo. Ukuba isampulu ifumana umbala obuthathaka kakhulu, tshintsha indlela ye-pH uze ukhethe ekaLoeffler. Ukuba yonke into ifumana umbala kodwa akukho nto yahlulwayo, tshintsha uhlobo lwedayi uze ukhethe ipolychrome. Ukuba isampulu eqatywe kwisilayidi yafakwa umbala ngekaLoeffler ibonisa iigranuli kakuhle kodwa umphathi ubuza ukuba kutheni iinucleus necytoplasm zinombala ofanayo oluhlaza okwesibhakabhaka, impendulo kukuba akukho dayi enealkali yombala omnye enokwenza umsebenzi wokuzahlula ngemibala. Loo mbuzo ngowosapho lwepolychrome kunye needayi zegazi ezivela kulo.

Ukuphepha ukubhidanisa okuqhelekileyo

Iimpazamo ezimbini zibangela uninzi lokubhideka. Eyokuqala kukuphatha ekaLoeffler njengento enokutshintshiselwana nemethylene blue eqhelekileyo kuba zombini zibonakala ziluhlaza okwesibhakabhaka ebhotileni. Ziyahluka nge-pH nangobunzulu bombala, yaye ukufaka idayi ene-pH ephakathi endaweni yekaLoeffler apho ifunwa khona kulahlekelisa kanye umahluko weegranuli inkqubo eyayilungiselelwe ukuwuveza. Eyesibini kukuphatha nayiphi na idayi eluhlaza okwesibhakabhaka enealkali endala okanye egcinwe ixesha elide ngempazamo njengepolychrome. Ukugcinwa ixesha elide okungalawulwayo kuvelisa umxube ongaziwayo onamandla angaziwayo, ngoxa izisombululo zokwenene zepolychrome zilungiswa ngeendlela zeoksideshini ezichaziweyo yaye, kusapho lweedayi zegazi, zidityaniswa ne-eosin elinganisiweyo. Imbali ifundisa apha: izisombululo ezivuthisiweyo zasemanzini ezingazinzanga zeminyaka yee-1890 zathatyathelwa indawo yimixube ezinzisiweyo yemethanol kanye ngenxa yokuba imixube engalawulwayo yayingenakwenziwa kwakhona ngendlela efanayo, isifundo ekufanele sikhunjulwe ngaphambi kokufaka umbala kwisilayidi sokuxilonga usebenzisa ibhotile engaziwayo ukuba indala kangakanani.

Ukuchaneka kwamagama kugqibezela oku. Qinisekisa imveliso ekwikhathalogu (eyasemanzini, ekaLoeffler okanye eyepolychrome), isixa sedayi, nendlela ecetyelwe ukusetyenziswa ngayo kwileyibhile, kunokuba uqikelele ngombala. Le mixube mithathu ivela kwisiseko esinye sechromophore. Ayinanjongo inye.