Isihloko
Ubani owasungula udayi onsomi wokufaka umbala egazini? I-methylene blue, uMalachowski, uRomanowsky noGiemsa
I-eosin ibomvu. I-methylene blue iluhlaza okwesibhakabhaka. Hlanganisa lezi ezimbili, ufake ingxube ongqimbeni oluncane lwegazi olomisiwe, bese ezinye izingxenye zalolo ngqimba ziba nsomi.
Akuzona zonke izingxenye. Esampuleni elungiselelwe kahle, ama-nucleus amangqamuzana amhlophe egazi aba nombala obomvu ngokunsomi, kanti ngaphakathi kwesilokazane sikamalaleveva i-chromatin iba nombala ofanayo, kuyilapho amangqamuzana abomvu azungezile ehlala ebomvana futhi i-cytoplasm ihlala impunga ngokuluhlaza okwesibhakabhaka. Umbala onsomi yiwo owenza kube lula ukuhlukanisa izinhlobo zamangqamuzana.
Alikho kulawa mabhodlela eliqukethe umbala onsomi. Lo mbala uvela kuphela lapho umkhiqizo othile wokubola kwe-methylene blue uhlangana ne-eosin engilazini yesibonakhulu, futhi ngeminyaka yawo-1880 neyawo-1890 akekho owayazi ukuthi lowo mkhiqizo ukhona, ingasaphathwa eyokuthi uyini. Abantu ababelandela lo mbala babesebenza ngodayi abangahlanzekile, bebodwa, bengenayo indlela ethembekile yokuthola ukuthi yini eyayisebhodleleni ngemva kokuhlala eshalofini izinyanga ezimbili.
Yingakho umbuzo osesihlokweni unezimpendulo ezingaphezu kweyodwa ezingasekelwa, futhi yingakho impendulo yashintsha ngo-2011.
Okwakubonakala kwakushintsha nengxube
Lo dayi awusungulwanga ngesikhathi esisodwa. Wathuthuka kancane kancane. Ushintsho ngalunye engxubeni lwaveza into eyodwa entsha ebonakalayo, futhi ukulandelana kwalezo zinto kuyindlela ecace kakhulu yokufunda lo mlando.
| Unyaka | Okwakusesitsheni | Okwakubonakala |
|---|---|---|
| 1865 | I-picric acid, udayi wokuqala owenziwe ngokwenziwa owafakwa egazini, nguWilhelm Erb | Iphethini efana nendwangu yeleyisi ngaphakathi kwamanye amangqamuzana abomvu. |
| 1876 | I-methylene blue, eyenziwa nguHeinrich Caro noAdolf von Baeyer kwa-BASF | Amagciwane esifo sofuba amise okwezinti, lapho esetshenziswa nguRobert Koch. Amangqamuzana egazi anomthunzi owodwa oluhlaza okwesibhakabhaka. |
| 1880 | I-methylene blue ne-acid fuchsin, udayi ongathathi hlangothi ka-Ehrlich | Amangqamuzana abomvu nezinhlamvana zama-eosinophil kubomvu, ama-nucleus aluhlaza okwesibhakabhaka ngokujulile, i-cytoplasm yama-lymphocyte iluhlaza okwesibhakabhaka ngokuphaphathekile, nezinhlamvana ezinsomi ngokumnyama ezimunca bobabili odayi. Yingakho leli ngqamuzana libizwa ngokuthi i-neutrophil. Ukubalwa kokuqala kwamangqamuzana amhlophe ngokwezinhlobo zawo. |
| 1888 | I-methylene blue ne-eosin. UCzesław Chęciński | Isilokazane uqobo: izimo eziluhlaza okwesibhakabhaka ezifana nembali ye-daisy (Plasmodium malariae) nezimo ezifana nesikela (ama-gametocyte e-P. falciparum). Hhayi i-nucleus yaso. |
| 1890 | I-methylene blue eyengeziwe, utshwala obuncane, kanye ne-potassium hydroxide. U-Albert Plehn | Uhlu lwemibala olubanzi nolujulile, kuhlanganise namangqamuzana abomvu nezinhlamvana zama-eosinophil ezinombala we-rose. Kwakusengobomvu noluhlaza okwesibhakabhaka kuphela. |
| 1890 | I-eosin ne-methylene blue eyenziwe yaba ne-alkali nge-borax. U-Ernst Malachowski | I-chromatin ebomvu ngokunsomi ngaphakathi kwesilokazane, i-nucleus ihlukaniseka ne-cytoplasm. Kwenziwa ngenhloso, futhi ngokombiko wakhe kwakuphindeka. |
| 1891 | I-eosin ne-methylene blue evela ebhodleleni elinesikhunta elalinezinyanga ezingaphezu kwezimbili. UDmitri Romanowsky | Umbala ofanayo onsomi, kanye kuphela, ngendlela abanye ababengakwazi ukuyiphinda. |
| 1898 kuya ku-1902 | Udayi onemibala eminingi okhishwe nge-chloroform (uNocht); i-methanol (uJenner); ukuncibilikisa kabusha inhlabathi eyakhekile ngokusebenzisa utshwala (uLeishman, uReuter, uWright); odayi abalinganisiwe abane-glycerol (uGiemsa) | Umbala onsomi noma nini lapho udingeka, emkhiqizweni owawusebenza ngendlela efanayo kwelinye ilabhorethri. Inguqulo kaJenner, eyayingenawo udayi oguqulwe waba nemibala eminingi, ayizange ikhiqize umbala onsomi. |
| 1940 kuya ku-1943 | ULillie, uRoe noWilcox bahlukanisa izingxenye bazihlola ngayinye ngayinye | I-Azure B, hhayi ingxube iyonke, iyingxenye ekhiqiza umbala onsomi. |
Iphethini esekholomini yokugcina yiyona ebalulekile. Yonke inqubekela phambili yandisa uhlu lwemibala, futhi ukwanda ngakunye kwenza olunye uhlaka lubonakale. Umbala onsomi awuwona umhlobiso wale ndlela; yiwo owenza umehluko phakathi kokubona ukuthi kukhona isilokazane nokubona ukuthi sakhiwe ngani. Ukuthi udayi uqobo wasuka kanjani embonini kadayi ye-BASF waya kwezokwelapha kungumlando omude onemibhalo yawo. Konke ukulandelana okungenhla, kanye nomthombo wesigaba ngasinye, kugcinwe ohlelweni lwezikhathi lwabahlanganyeli oluxhumanisa nemithombo.
Umdwebo: zona lezi zingxube eziyisithupha zidwetshwe zibonisa okwakusesitsheni nalokho okwavela kuso. Imibuthano ibonisa odayi; amabhulokhi angaphansi abonisa imibala labo dayi ababengayikhiqiza ongqimbeni lwegazi. Akukho ngaphambi kuka-1890 okubonisa ibhulokhi elibomvu ngokunsomi, futhi akukho ngemva kwakhe okungalibonisi.
Indoda eyasebenzisa i-borax ngenhloso
U-Ernst Malachowski wayengudokotela eBreslau, manje ebizwa ngeWrocław, owachitha umsebenzi wakhe wezobudokotela esebenza ngokuzimela nasemtholampilo omncane ayewuphethe nozakwabo, enza umsebenzi waselabhorethri ngesikhathi sakhe sokuphumula. Wazalelwa eStrzelno mhla zi-12 kuNdasa 1857, wafunda ubudokotela eNyuvesi yaseBreslau, wasebenza kusukela ngo-1882 kuya ku-1887 njengomsizi emkhakheni wezifo zangaphakathi esibhedlela i-All Saints Hospital, esasiyisibhedlela sesibili ngobukhulu eJalimane ngaleso sikhathi.
Waqala ngo-1888 ngokusebenzisa odayi ababili bakaChęciński noPlehn. Esampuleni ayeyilungiselele ngo-1889 wabona i-chromatin ye-nucleus ye-plasmodium iba nombala ohlukile kowe-cytoplasm, kodwa wabe esethola ukuthi akakwazi ukuphinda lowo mphumela. Wazama ukwenza i-methylene blue ibe ne-asidi, okwenza izinto zaba zimbi kakhulu. Wazama ukuyenza ibe ne-alkali nge-borax, okwakusebenza njalo. Wabonisa le ndlela emphakathini mhla zi-15 kuNhlangulana 1890 futhi washicilela incazelo eningiliziwe ngoNcwaba 1891, ehlanganisa nomyalelo wokusebenzisa i-borax. Waphawula ukuthi ukufakwa kombala kwakuthatha amahora angamashumi amabili nane nokuthi imiphumela yayiphindeka kalula.
Amahora angamashumi amabili nane yisikhathi eside elabhorethri yokuxilonga, futhi iphepha lakhe alizange ligxile embala onsomi ngokubomvu we-nucleus yesilokazane. Noma ngabe kwakuyini isizathu, akekho owasebenzisa ukwenza i-methylene blue ibe ne-alkali ngenhloso kwaze kwaba yilapho uNocht ephinda ekwenza ngo-1898. UMalachowski akaphindanga wabhala ngokufaka umbala egazini.
Isizathu sokuthi kungani kwakuncane kangaka okwakwaziwa ngaye kuze kube muva nje simayelana nezinqolobane zemibhalo, hhayi imfihlakalo. Cishe akukho mkhondo wokuphila kwakhe owasala emibhalweni yezokwelapha, futhi lokho okwasala kwafika ngomzila owodwa. Indodakazi yakhe uMarie yasuka eJalimane yaya eLondon ngo-1939, yasebenza ebuthweni laseBrithani, yabe isizinza eCanada, futhi iqoqo layo lemibhalo yomndeni lagcina liseLeo Baeck Institute eNew York. Umfowabo uLeo wabulawa eMpini Yezwe Yokuqala. Udadewabo u-Eva wasuswa ngenkani eBreslau wayiswa enkambini yokudlulisa iziboshwa yase-Izbica ngoMbasa 1942, futhi akazange abuye. Unina uHedwig wazibulala mhla zingama-22 kuMbasa 1942 kunokuba alinde ukususwa ngenkani ngendlela efanayo. UMalachowski wayeshone ngo-1934, ngaphambi kwakho konke lokho.
Umlando womndeni ophela kanjalo uba umlando ongabe usacashunwa. Amaphepha amabili abuyisela uMalachowski emlandweni wokuthuthukiswa kwalo dayi, womabili ashicilelwa ngo-2011, ahlanganiswa kusetshenziswa imibhalo yezinqolobane zasePoznań naseWrocław kanye nemibhalo kaMarie eNew York.
Ibhodlela elinesikhunta nomRussia owaba nenhlanhla
UDmitri Romanowsky wafunda ubudokotela eMilitary Medical Academy eSt Petersburg futhi wasebenza ngomalaleveva isikhathi esiningi somsebenzi wakhe. Ngo-1890 washicilela indlela eyayicishe ifane nekaChęciński ngokwalokho okwakubhaliwe: i-methylene blue engaguquliwe ne-eosin, kodwa ene-eosin ethe xaxa.
NgoMandulo 1891 washicilela indlela eyamenza waduma, futhi umniningwane owawubalulekile wawufihlwe phakathi kwemiyalelo. Imiphumela yakhe emihle kakhulu yayivela ku-die vorräthige Methylenlösung, isixazululo se-methylene blue esasigcinelwe ukusetshenziswa futhi esasihleli izinyanga ezingaphezu kwezimbili sikhula isikhunta. Isikhunta nesikhathi sokuhlala kwakususe amanye amaqembu e-methyl kudayi. URomanowsky wayengakwazi lokho futhi engakwazi ukukulawula, kanti abanye abasebenzi abazama indlela yakhe abakwazanga ukuthola umbala onsomi. Umbhalo wakhe wocwaningo wayethunyelwe eSt Petersburg ngoNhlangulana 1891, yingakho umlando ojwayelekile uthatha u-1891 njengonyaka wokutholakala kwalokhu.
UMalachowski wayebonise indlela yakhe ngoNhlangulana 1890, ngaphezu konyaka ngaphambili, futhi indlela yakhe yayisebenza ngenxa yesinyathelo esasenziwa ngenhloso.
Kungani igama lendlela laba elendoda okungeyona
Ubufakazi obusekela uMalachowski bahlanganiswa nguKristine Krafts, u-Ernst Hempelmann noBarbara Oleksyn, futhi busekelwe ezintweni ezine. Usuku lokubonisa indlela emphakathini: uNhlangulana 1890 uma kuqhathaniswa noMandulo 1891. Igama elithi ngenhloso uma liqhathaniswa nelithi ngenhlanhla. Ukuthi uMalachowski washicilela indlela yakhe, kuhlanganise ne-borax, kuyilapho ekaRomanowsky yayincike entweni engalawulwa. Nokuthi incazelo ye-nucleus ye-plasmodium, kanye nombala onsomi owawuyiveza, cishe yanyamalala emibhalweni iminyaka eyisikhombisa, ngoba akekho owayekwazi ukuphinda inguqulo eyayenzeke ngengozi.
Abazungezi amagama ngesiphetho sabo. Ephepheni lomlando wakhe wokuphila babiza lokhu kwabelwa kodumo ngokuthi “ukuhlanekezela okubi”, futhi bacaphuna uRalph Lillie, owabhala ngo-1978 ukuthi “mhlawumbe kufanele sibize leli qoqo lodayi ngokuthi odayi bakaMalachowski”.
Lesi simangalo siyaphikiswa, njengoba kuvame ukwenzeka ezimangalweni zokuthi ubani owaqala, futhi uhlobo lwale mpikiswano kufanele luchazwe kunokuba lunqunywe. Impikiswano yokuthi ubani owaqala kufanele ichaze ukuthi yini eyatholwa. Uma okwatholwa kungumbala onsomi, uMalachowski unobufakazi obubhaliwe bokuwubonisa ngaphambili. Uma okwatholwa kuyindlela eyashicilelwa, bobabili bashicilela ngo-1891 futhi indlela yaqanjwa ngoRomanowsky. Uma okwatholwa kuyincazelo, akekho kubo onelungelo lokuzibiza ngomtholi, ngoba akekho owayazi ukuthi ukuvuthisa kwakuwukususa amaqembu e-methyl, futhi akekho owayengasho ukuthi kungani lo mbala uvela. Leli phuzu lokugcina yilona elishiywa yizincazelo ezithandwayo. Ingxenye esebenzayo kwakuwukungcola okwakungakahlonzwa muntu, futhi umuntu wokuqala owakhiqiza umbala ngenhloso wakwenza ngokulandela indlela eyayiphindeka esekelwe ekuhloleni, hhayi ekuqondeni lokho le ndlela eyayikwenza.
Amaphepha amabili ango-2011 ayizincazelo ezishicilelwe eziphelele kakhulu zobufakazi obusekela uMalachowski, futhi womabili afundeka mahhala ku-PubMed: ukubuyekezwa okude ku-PMID 21235291 nephepha lomlando wakhe wokuphila ku-PMID 21660627. Irekhodi le-PubMed lalokhu kubuyekezwa libuye libale ukuphawula ngakho, okwashicilelwa ephephabhukwini elifanayo kamuva ngalowo nyaka ku-86(5):366, okuyisikhumbuzo esifanele sokuthi le mpikiswano yaphikiswa emibhalweni ngokushesha lapho ivela.
Le mpikiswano ayiphathelene nokungavumelani ngamakhemikhali. Bonke ababandakanyekayo bayavumelana ngokuthi umbala onsomi uyini nokuthi yiziphi izingxenye eziwukhiqizayo. Ukungavumelani kumayelana nokuthi ubani owafika lapho kuqala, nokuthi ingabe isenzakalo esenzeka ngengozi esasingaphindeka muntu singabalwa njengokufika lapho nhlobo.
Okwafezwa amagama akamuva
Uma uMalachowski ekhiqiza umbala onsomi kuqala, kungani udayi ubizwa ngamagama kaRomanowsky, kaWright nakaGiemsa?
Ngoba udayi osebenza kanye ezandleni zomsunguli wawo awukabi yisivikeli esithembekile sokuhlola. Umsebenzi wakamuva waguqula ingxube eyayingathembeki yaba into uchwepheshe wakwelinye izwe ayengayisebenzisa ngoLwesibili.
UNocht wafinyelela kulokho ngokucabangisisa. Ngemva kokwehluleka ukuthola umphumela othembekile kudayi olula ongathathi hlangothi, waphetha ngokuthi umbala onsomi kufanele uvele entweni ese-methylene blue engeyona i-methylene blue, wabe eseqala ukuyifuna. U-Unna wayesebonile ukuthi izixazululo ze-methylene blue ezasezindala zazifaka umbala obomvu ezinhlamvaneni zama-mast cell, futhi wasikisela ukuthi kwakheka into entsha njengoba udayi uvuthiswa. UBernthsen wayebonise ngo-1885 ukuthi izimo ezine-alkali zisusa amaqembu e-methyl ku-methylene blue, nokuthi le mikhiqizo iyilokho manje esikubiza ngama-azure. UNocht wakhipha ingxenye esebenzayo nge-chloroform futhi wakwazi ukuyibiza kuphela ngokuthi Rot aus Methylenblau, okubomvu okuvela ku-methylene blue; uMichaelis wayiqamba ngokuthi i-azure I ngo-1901. UJenner waxazulula izinkinga zokulungiselela nokusebenzisa. Ukuncibilikisa kabusha inhlabathi eyakheka nge-methylene blue ne-eosin ku-methanol engenamanzi kwakhiqiza isixazululo esisodwa esizinzile esasimisa ungqimba lwegazi ngesikhathi silufaka umbala, futhi saqeda isidingo sokulumisa ngokushisa. ULeishman noReuter bahlanganisa udayi oguqulwe waba nemibala eminingi nokuncibilikisi okusekelwe otshwaleni, bathola kokubili ukuzinza nombala onsomi. UWright washintsha ukufukanyelwa kwamahora ayishumi nambili ku-65 °C kanye nokuhlala izinsuku eziyishumi, esebenzisa ihora elilodwa esitimu esigelezayo esikhundleni salokho. UGiemsa wayeka ngokuphelele ukuthembela esixazululweni esivuthiswe ngokungahleliwe, wasebenzisa amanani alinganisiwe odayi abaziwayo ne-glycerol ku-methanol ukuze umkhiqizo umelane nokushisa kwasezindaweni ezishisayo. Akazange ashicilele ukuthi wayenza kanjani i-methylene azure ehlanzekile. Wayithengisela ilabhorethri kaDkt Grübler eLeipzig, eyayimaketha njenge-azure I futhi ithi inguqulo yayo kuphela eyayiwudayi wangempela kaGiemsa.
Ukuthi leyo azure yayiyini kusaphikiswana ngakho. Abanye bathi kwakumane kuyi-methylene blue eguqulwe yaba nemibala eminingi, abanye bathi yayine-Azure B eningi. Impendulo yavela ngenxa yempi. Lapho odayi baseJalimane bengasatholakali, ithimba le-United States Public Health Service, uRalph Lillie enoMaurice Roe no-Aimee Wilcox, lathola ukuthi ukufakwa kombala okuhle kakhulu kwakuvela ezingxenyeni ezintathu ezihlukanisiwe: i-methylene blue engaguquliwe, umkhiqizo wokubola kwayo osuswe amaqembu e-methyl, i-Azure B, kanye ne-eosin. I-Azure B yathatha indawo ye-azure I eyayingasatholakali, futhi lezi zingxenye ezintathu ziseyisisekelo sikadayi osetshenziselwa ukuxilonga umalaleveva namuhla.
Indlela lezi zingxenye ezintathu ezisebenzisana ngayo ngokwamakhemikhali engilazini yesibonakhulu ingumbuzo ohlukile futhi isekhasini elihlukile. Ikhasi lezindima zodayi namakhemikhali odayi bohlobo lukaRomanowsky lichaza ukuthi i-Azure B ne-eosin ngayinye yenzani nokuthi kungani lezi ezimbili zikhiqiza umbala okungekho neyodwa ewukhiqizayo iyodwa. Ikhasi lokusetshenziswa kwezibonakhulu nokwaselabhorethri lichaza uhlangothi lokusebenza: ukuthi isampula ethile elungiselelwe ibonisani, nokuthi ungahlulela kanjani ukuthi umkhiqizo othile uyifanele yini indlela ohlose ukuyisebenzisa.
Ingxenye yomlando esasasebenza nanamuhla
Umniningwane owodwa kule ndaba awuzange uyeke ukuba yiqiniso ngo-1902. Cishe iminyaka engamashumi amahlanu, ingxenye ebaluleke kakhulu kadayi ojwayelekile womhlaba wokuhlola umalaleveva yayiwukungcola okungenagama. Yayikhona ngengozi, ubungako bayo buhlukahluka emabhodleleni, ukuthi iyini kwakuyimfihlo yezohwebo ingxenye yaleso sikhathi, futhi kwakuphikiswana ngesilinganiso sayo kuze kube yiminyaka yawo-1940. Abantu ababesebenzisa lo dayi babenganaki. Babengenayo indlela yokwazi ukuthi yini eyayisebhodleleni.
Uhlobo lwanamuhla lwale nkinga efanayo luwumbandela ochaza ingxenye kuphela yento. Ukuhambisana ne-USP kuyindlela yokufinyelela ekuhlanzekeni okuphezulu kakhulu okutholakala emakethe, ngoba iyindinganiso ephelele eshicilelwe kunokuba ibe yisilinganiso esisodwa: ukuhlonzwa kwento, ukuhlanzeka, ukungcola kwamakhemikhali emvelo okuhlanganisa odayi be-azure, izincibilikisi ezisele, ukungcola kwezakhi, insalela ngemva kokushiswa, imikhawulo yamagciwane nama-endotoxin amagciwane. I-Blupreme ithola i-methylene blue yayo kumkhiqizi wemithi futhi ishicilela isitifiketi esiphelele sokuhlaziywa, ukuze wonke umugqa walolo hlu uhlolwe kunokuba kushiwo nje ukuthi uhlangatshezwana nawo. Isitifiketi esibika umphumela wezinsimbi ezisindayo bese sibiza lokho ngokuhambisana ne-USP siyafana nanamuhla nebhodlela elibhalwe ukuthi “i-methylene blue evuthisiwe”: kungenzeka kube yikhemikhali enhle yokuhlola, kodwa ilebula ayikutsheli ukuthi yini ekulo.