Immunostaining

I-Immunostaining bubuchule obubalulekileyo baselabhoratri obusetyenziswa kwibhayoloji yemolekyuli kunye ne-pathology ukubona ubukho beeproteni ezithile okanye ii-antigens kwiisampulu zethishu. Ngokusebenzisa amajoni omzimba ukuchonga ezi thagethi, i-immunostaining ivumela izazinzulu kunye neengcali zonyango ukuba ziqonde iimpawu zezifo ezahlukeneyo, kubandakanya umhlaza, ukuphazamiseka kwe-autoimmune, kunye nezifo ezosulelayo. Le ndlela iluncedo kakhulu kwi-diagnostic pathology, ukunceda oogqirha baxilonga izifo ngokuchanekileyo, umhlaza wesigaba, kunye nokubeka iliso kwiimpendulo kunyango.

Yintoni Immunostaining?

I-Immunostaining (ekwaziwa ngokuba yi-immunohistochemistry) bubuchule baselabhorethri obusetyenziselwa ukufumanisa ii-antigens ezithile (iiproteni) kwiiseli zecandelo lethishu ngokusebenzisa izilwa-buhlungu ezibophelela ngokukodwa kwezi antigens. Ezi zithinteli zidla ngokuphawulwa ngesiphawuli esibonwayo, esifana nedayi ye-fluorescent okanye i-enzyme evelisa utshintsho lombala xa iboniswe kwi-substrate. Oku kuvumela iingcali zezifo ukuba zibone ubukho beeprotheyini ezithile okanye iimpawu kwiisampulu zethishu phantsi kwemakroskopu.

I-Immunostaining isebenza kumgaqo wokusebenzisana kwe-antigen-antibody, apho i-antibody ibophelela kwi-antigen ethile, yenza i-antigen-antibody complex. Isiphawuli esincanyathiselwe kwi-antibody sivumela ukubonwa kolu bunzima.

Iimpawu eziphambili ze-Immunostaining

• Ukucaciswa: I-Immunostaining igxininise kakhulu njengoko ijolise kwiimpawu ezizodwa (iiprotheni) ezifumaneka kwiiseli okanye izicubu ezithile.
• Ukuqonda: Iyakwazi ukubona ixabiso eliphantsi kakhulu le-antigens, eyenza kube luncedo ekuchongeni izifo kwisigaba sokuqala.
• Isiphumo Esibonakalayo: Iziphumo zibonwa ngebala, elijongwa phantsi kwe-microscope. Oku kunika ii-pathologists ukuqonda okucacileyo kwisampulu yethishu.
• Multiplexing: Ubuchwephesha be-immunostaining obuphezulu bunokubona ii-antigens ezininzi ngaxeshanye, ezixabisekileyo ekuchongeni izifo ezintsonkothileyo okanye abamakishi.

Isebenza Njani I-Immunostaining?

Inkqubo ye-immunostaining ibandakanya amanyathelo amaninzi abalulekileyo:

1. Ukulungiswa kwezicubu: Isampulu yethishu, efana ne-biopsy okanye uqhaqho loqhaqho, iyaqokelelwa. Emva koko i-tissue ilungiswe kwisisombululo (ngokuqhelekileyo i-formalin) ukugcina isakhiwo sayo kunye nokunyaniseka. Xa sele ilungisiwe, i-tissue ifakwe kwiparafini kwaye ihlulwe ibe ngamaqhekeza amancinci.
2. Ukubuyiswa kweAntigen: Inkqubo yokulungiswa ngamanye amaxesha inokufihla i-antigens, ngoko ke amacandelo e-tissue aphathwa ukuze aveze iiprotheni. Le nkqubo ibizwa ngokuba yi-antigen retrieval, enokuthi ibandakanye ukufudumeza okanye ukusebenzisa iikhemikhali ezikhethekileyo ukutyhila ii-antigens.
3. Ukusetyenziswa kwee-Antibody: Emva kokufunyanwa kwe-antigen, i-antibody ethile isetyenziswa kwi-tissue. Le antibody ibophelela kwi-antigen yomdla. Kwimeko yomhlaza, umzekelo, izilwa-buhlungu ezithile zingasetyenziselwa ukubophelela kwiimpawu ezifana ne-HER2 (yomhlaza wamabele) okanye i-PSA (yomhlaza wedlala lesinyi).
4. Ukufumanisa: Emva kokuba i-antibody ibophelela kwi-antigen, i-antibody yesibini ifakwa. Le antibody yesibini idla ngokudityaniswa kwi-enzyme okanye idayi ye-fluorescent. I-enzyme iphendula kunye ne-substrate ukuvelisa utshintsho lombala, olunokubonwa phantsi kwe-microscope. Kwi-fluorescence-based immunostaining, idayi ibangela i-fluorescence phantsi kokukhanya kwe-ultraviolet (UV), inika umfanekiso ocacileyo nochanekileyo wendawo ye-antigen.
5. Uhlalutyo: I-tissue enebala emva koko ihlolwe ngabagqirha bezifo ezitolika iziphumo ngokusekelwe kwipatheni yebala, ukuqina, kunye nendawo.

Kutheni I-Immunostaining Yenziwa?

I-Immunostaining inezicelo ezininzi ezibalulekileyo kuzo zombini uxilongo lwezonyango kunye nophando. Ezinye zezona zizathu ziqhelekileyo zokwenza i-immunostaining ziquka:

• Ukuxilongwa koMhlaza kunye noHlelo: Enye yezona zinto zibalulekileyo zokusetyenziswa kwe-immunostaining kwi-oncology. Inceda ekuxilongeni iintlobo ezahlukeneyo zomhlaza ngokuchonga iimpawu ezithile ezikhoyo kwiiseli zethumba. Umzekelo, i-HER2 yiprotheyini efunyenwe ngokugqithisileyo kwezinye iiseli zomhlaza wamabele, kwaye ukuchongwa kwayo ngonyango lwe-immunostaining kunokugqiba iindlela zonyango. I-Estrogen kunye ne-progesterone receptors nazo zichongiwe kwiithishu zomhlaza webele ukuze ziqinisekise ukuba unyango lwe-hormone lunokusebenza.
• Ukuchonga izifo ezosulelayo: I-Immunostaining isetyenziselwa ukuchonga iintsholongwane ezinjengeentsholongwane, iintsholongwane, okanye umngundo kwiisampulu zethishu. Le ndlela inokunceda ekubhaqeni i-HIV, isifo sephepha, kunye nezinye ii-agent ezosulelayo kwiisampuli ze-biopsy. Ukukwazi ukujonga ngokuthe ngqo i-pathogen kwi-tissue kubaluleke kakhulu ukuxilongwa ngokuchanekileyo kunye nonyango.
• Uxilongo lweSifo somzimba: I-Immunostaining isetyenziselwa ukufumanisa i-autoantibodies ethile kunye nokuchonga izifo ezizimelayo, ezifana ne-lupus kunye ne-rheumatoid arthritis. Ngokomzekelo, kwi-systemic lupus erythematosus (SLE), i-immunostaining inokunceda ukuchonga ama-antibodies abopha kwiiprotheni ezithile kwiintso, ezinokubonisa i-lupus nephritis.
• Ukuhlalutya uLwakhiwo lweTishu: I-Immunostaining iphinda incede ukuhlalutya isakhiwo kunye nokubunjwa kwezicubu. Oku kubalulekile ekuqondeni izifo ezichaphazela ukwakhiwa kweeseli, ezifana nezifo ze-neurodeergenerative okanye izifo ze-vascular. Isetyenziselwa ukufunda i-matrix ye-extracellular kunye neeprotheni ezigcina ukwakheka kwezicubu.
• Ukubeka iliso ekuqhubeleni phambili kweSifo kunye neMpendulo yoNyango: Kwezinye izifo, ezifana nomhlaza okanye ukuphazamiseka kwe-autoimmune, i-immunostaining isetyenziselwa ukuvavanya ukuqhubela phambili kwesi sifo. Umzekelo, i-immunostaining inokubeka iliso ekubonakalisweni kweempawu ezithile zethumba ukumisela ukuba isigulana siphendula kangakanani na kunyango.

Indlela yokuLungiselela i-Immunostaining

Akukho malungiselelo akhethekileyo esigulane afunekayo ekuzixhomeni ngokwaso, njengoko bubuchule obusekwe kwilabhoratri esetyenziswa kwiisampulu zethishu. Nangona kunjalo, ukuba wenza i-biopsy okanye utyando lokuqokelela isampulu ye-immunostaining, kukho izinto ezimbalwa ekufuneka uzigcine engqondweni:

• Uthethwano kunye neMbali: Ngaphambi kokuba wenze i-biopsy okanye utyando, ugqirha wakho unokufuna ukuphonononga imbali yakho yezonyango ukuze abone ukuba zeziphi izilwa-buhlungu ekufuneka zisetyenziselwe ukungazinzi. Ukuba uphantsi kolu vavanyo ukuvavanya uhlobo oluthile lomhlaza okanye isifo, yazisa ugqirha wakho malunga nazo naziphi na iimpawu okanye imbali yosapho yeemeko ezinxulumeneyo.
• I-Biopsy okanye ukuqokelelwa kwezicubu: Ukuze kugcinwe i-immunostaining, i-biopsy okanye i-resection yoqhaqho ingafuneka ukufumana isampuli yezicubu. Ukuba i-biopsy yenziwe, umboneleli wakho wezempilo uya kuchaza inkqubo, enokuthi ibandakanye i-anesthesia yendawo, i-sedation, okanye i-anesthesia jikelele, ngokuxhomekeke kwindawo kunye nobukhulu besampulu yethishu.
• Ukuphatha iSampuli emva: Emva kokuba isampuli iqokelelwe, iya kuthunyelwa kwibhubhoratri, apho iya kucutshungulwa kwaye ilungiselelwe i-immunostaining. Akukho malungiselelo ongezelelweyo esigulane afunekayo emva kokuba isampulu yethishu ithathiwe.

Utoliko lweziphumo zovavanyo

Iziphumo ze-Immunostaining zitolikwa ngokusekwe kubukho kunye nokuqina kokungcola kwisampulu yethishu. Nantsi indlela yokutolika iziphumo:

• Iziphumo Ezilungileyo:
  • Ibala elilungileyo eliqinileyo: Ibonisa ubukho be-antigen ekujoliswe kuyo okanye iprotheni kwi-tissue. Ubunzulu bebala bunokunceda ukufumanisa ubuninzi be-antigen.
  • Ipateni eNgcolileyo yokuNgcolisa: Ipatheni apho amabala e-tissue (umzekelo, inyukliya, i-cytoplasmic, i-membranous) inokubonelela ngolwazi malunga nendlela i-antigen esebenza ngayo kwiseli.
• Iziphumo ezibi:
  • Akukho Kubala: Ukunqongophala kwebala kucebisa ukuba i-antigen ekujoliswe kuyo ayikho okanye ikhona ngezixa eziphantsi kakhulu. Iziphumo ezibi zinceda ukukhupha izifo okanye iimeko ezithile.
• Utoliko lweNdawo eSwala:
  • Ukungcola kweCytoplasmic: Ibonisa ukuba i-antigen ifumaneka kwi-cytoplasm yeeseli.
  • Ukungcoliswa kweNyukliya: Iphakamisa ukuba i-antigen ikhona kwi-nucleus yeseli, mhlawumbi ichaphazela i-gene expression.
  • Ukudyobheka kwiMembranous: Ibonisa ukuba i-antigen ikhona kwi-membrane yeseli, enokubaluleka kwiinkqubo ze-receptor-mediated.
• Ubungakanani: I-Immunostaining inokunceda ukumisela ukubonakaliswa komyinge we-antigens, inika ulwazi oluluncedo kuhlelo lwe-tumor, apho inqanaba lokubonisa eliphakamileyo linokubonisa ithumba elinobundlobongela.

Uluhlu oluqhelekileyo lwe-Immunostaining

Ekubeni i-immunostaining ichonga iiprotheyini ezithile, akukho "uluhlu oluqhelekileyo" oluqhelekileyo njengoko kukho uvavanyo lwegazi. Endaweni yoko, iziphumo eziqhelekileyo zisekelwe ekungabikho kweziphawuli ezingaqhelekanga kwisampulu yethishu. Ubukho beempawu ezithile okanye iiprotheni, ezifana ne-HER2 kumdlavuza webele okanye iimpawu ze-CD kwi-lymphoma, ziya kugqiba ukuba umphumo uyinto eqhelekileyo okanye ibonisa isifo.

Ukusetyenziswa kwe-Immunostaining

I-Immunostaining idlala indima ebalulekileyo ekuxilongeni nasekulawuleni iimeko ezahlukeneyo zonyango:

• Ukuxilongwa koMhlaza: Isetyenziselwa ukuchonga iimpawu ze-tumor kunye nokuhlelwa komhlaza, njengomhlaza webele, umhlaza wesifo seprotate, kunye ne-lymphomas.
• Izifo ezosulelayo: Inceda ukufumanisa iintsholongwane ezifana neentsholongwane, ibhaktheriya, kunye nomngundo kwiisampulu zethishu.
• Iziphazamiso ze-Autoimmune: Isetyenziselwa ukuchonga izifo ze-autoimmune kunye nokuxilongwa kwezifo ezizimele.
• Uphando: Isetyenziswa kakhulu kuphando lokufunda iindlela zesifo, ukuchonga iithagethi zonyango, kunye nokuphuhlisa unyango olutsha.
• Ukumila kwezicubu: Isetyenziselwa ukuvavanya izakhi zezicubu, ukunceda ekufundeni izifo ze-vascular, iimeko ze-neurodeergenerative, kunye nokungasebenzi kakuhle kwamalungu.

I-10 yemibuzo ebuzwa rhoqo malunga ne-Immunostaining

1. Yintoni Immunostaining? I-Immunostaining bubuchule baselabhoratri obusetyenziselwa ukubona iiproteni ezithile kwiisampulu zethishu ngokusebenzisa izilwa-buhlungu. Ezi zithinteli zibophelela kwi-antigen ekujoliswe kuyo, kwaye ukusabela kubonwa phantsi kwe-microscope.
2. Isebenza njani i-Immunostaining? I-Immunostaining isebenza ngokusebenzisa izilwa-buhlungu ezibophelela ngokukodwa kwiiprotheni (ii-antigens) kwisampulu yethishu. Ezi antibodies ziphawulwe ngeziphawuli ezivelisa ukusabela okubonakalayo, njengokutshintsha kombala okanye i-fluorescence.
3. Ziziphi iimeko ezinokuxilonga i-Immunostaining? I-Immunostaining isetyenziselwa ukuxilonga umhlaza, njengomhlaza webele, umhlaza wemiphunga, kunye ne-lymphoma. Ikwasetyenziselwa ukuxilonga izifo ze-autoimmune ezifana ne-lupus kunye ne-rheumatoid arthritis, kunye nokufumanisa usulelo.
4. Ichaneke kangakanani i-Immunostaining? I-Immunostaining ichanekile kakhulu kwaye inika ulwazi oluthile malunga nobukho bamanqaku athile kwiithishu, ukunceda ii-pathologists ngokuchanekileyo ukuxilonga izifo kunye nokuhlelwa komhlaza.
5. Ngaba i-Immunostaining ibuhlungu? I-Immunostaining ngokwayo ayibuhlungu kuba yenziwa kwiisampulu zethishu elabhoratri. Nangona kunjalo, ukufumana isampuli (umzekelo, ngokusebenzisa i-biopsy) kunokubandakanya ukungonwabi okuncinci.
6. Loluphi uhlobo lwesampulu olufunekayo kwi-Immunostaining? Isampulu yethishu iyafuneka ukuze kugcinwe amajoni omzimba, edla ngokufunyanwa nge-biopsy okanye inkqubo yotyando. Iisampulu zegazi okanye zolwelo zingasetyenziswa kwezinye iimeko.
7. Sithetha ntoni isiphumo esihle se-Immunostaining? Isiphumo esihle sithetha ukuba i-antigen ekujoliswe kuyo okanye iprotheni ifunyenwe kwisampuli yezicubu. Oku kunokunceda ukuxilonga iintlobo ezithile zomhlaza okanye ukuchonga iimpawu ezinxulumene nezifo ezithile.
8. Isiphumo esibi se-Immunostaining sithetha ntoni? Isiphumo esibi sithetha ukuba i-antigen ayizange ibonwe kwizicubu, ezinokukunceda ukulawula izifo okanye iimeko ezithile.
9. Kuthatha ixesha elingakanani ukufumana iziphumo ze-Immunostaining? Iziphumo ze-immunostaining zithatha iintsuku ezimbalwa ukuya kwiveki, ngokuxhomekeke kubunzima bovavanyo kunye nenani labamakishi abavavanywayo.
10. Ngaba i-Immunostaining ingasetyenziselwa izigqibo zonyango? Ewe, i-immunostaining ibalulekile ekumiseleni iindlela zonyango, ngakumbi kumhlaza. Umzekelo, ubukho be-HER2 kumhlaza webele bunokuqinisekisa ukuba isigulana siya kuxhamla kunyango olujoliswe kwi-HER2.

isiphelo

I-Immunostaining sisixhobo esiyimfuneko sokuxilonga kwiyeza, inceda ukufumanisa iiprotheyini ezithile kunye nee-antigens kwiisampulu zethishu. Ukusetyenziswa kwayo okubanzi ekuxilongeni umhlaza, izifo ezizimelayo, usulelo, kunye nophando lwenza ukuba ibe yindlela ebalulekileyo yeengcali zezifo kunye nabaphandi. Ingaba isetyenziselwa ukuguqulwa komhlaza, ukuxilongwa kwesifo somzimba, okanye ukujonga iimpendulo zonyango, i-immunostaining inikezela ngolwazi olubalulekileyo olukhokela izigqibo zonyango kunye nokuphucula iziphumo zesigulane.

Ngokukwazi kwayo ukuchonga amanqanaba aphantsi eeprotheyini ekujoliswe kuzo, i-immunostaining ayisosixhobo sokufumanisa izifo kuphela kodwa nokuqonda iindlela zeemolekyuli kunye neeseli ezisemva kwazo. Ukuba uphantsi kwe-immunostaining okanye ulungiselela i-biopsy ehambelanayo okanye uvavanyo lwezicubu, ukuxoxa ngenkqubo kunye nomboneleli wakho wezempilo kuya kunceda ukuqinisekisa ukuba uyaziqonda iingenelo ezinokuthi zibekho kunye nendlela iziphumo eziya kuchaphazela ngayo unyango lwakho.