Immunohistochemistry

I-Immunohistochemistry (IHC) bubuchule belebhu obunamandla obusetyenziselwa ukufumanisa ii-antigens ezithile (iiproteni) kwiiseli zecandelo lethishu. Idibanisa imigaqo ye-immunology kunye ne-microscopy ukuchonga kunye nokufumana iiprotheyini ezithile kwiisampulu zethishu, iyenza ibaluleke kakhulu kwiindawo ezininzi zophando lwezonyango, uxilongo, kunye ne-pathology. Ubuchule busetyenziswa ngokubanzi kuxilongo lomhlaza, kunye nokuchonga imvelaphi yethishu yezifo ezithile, ukunceda ekuqondeni ukuziphatha kwethumba kunye nezigqibo zonyango ezikhokelayo.

Yintoni i-Immunohistochemistry?

I-Immunohistochemistry (IHC) yindlela yaselabhoratri esetyenziselwa ukufumanisa ubukho bee-antigens ezithile kwiiseli zecandelo lethishu. Isebenzisa umgaqo wokusebenzisana kwe-antigen-antibody ukubopha ama-antibodies kuma-antigens, okuvumela ukuba i-antigen ibonakale phantsi kwe-microscope.

Inkqubo ye-IHC ibandakanya ukusebenzisa ii-antibodies eziphawulwe ngophawu olubonakalayo, njenge-enzyme okanye idayi ye-fluorescent, ukujolisa kunye nokubophelela kwii-antigens ezithile kwisampuli yezicubu. Ezi zithinteli-gazi zichane kakhulu kwi-antigen eyilelwe ukuyibona, zenza i-IHC ibe yindlela echane kakhulu yokuchonga iiproteni ezithile okanye iziphawuli kwiisampulu zethishu.

Isebenza njani i-Immunohistochemistry?

Amanyathelo asisiseko abandakanyekayo kwi-Immunohistochemistry ngala alandelayo:

1. ULungiselelo lweSampuli yezicubu: Isampuli yethishu (ngokuqhelekileyo isuka kwi-biopsy) iqokelelwa kwaye ilungiswe ukugcina isakhiwo sayo. Isampulu inqunyulwa ibe ngamacandelo amancinci kwaye ibekwe kwisilayidi seglasi.
2. Ukubuyiswa kweAntigen: Izicubu zihlala ziphathwa ngobushushu okanye iikhemikhali ukuze ziveze iiprotheyini okanye ii-antigens kwii-antibodies, njengoko ukulungiswa kwe-formaldehyde kunokufihla iisayithi ze-antigen.
3. Ukusetyenziswa kwe-Antibody: Icandelo le-tissue lifakwe kunye ne-antibody ephambili ebophelela ngokukodwa kwi-antigen ekujoliswe kuyo.
4. Ukubophelela kwe-Antibody yesibini: I-antibody yesibini iye isetyenziswe, ethi ibophelele kwi-antibody yokuqala kwaye idityaniswe ngesiphawuli (esifana ne-enzyme okanye idayi ye-fluorescent).
5. Ukufumanisa: Izicubu ziphathwa nge-substrate ephendula nge-enzyme okanye umakishi, ukuvelisa utshintsho lombala okanye i-fluorescence kwindawo ye-antigen. Oku kuvumela i-antigen ukuba ibonwe phantsi kwe-microscope.
6. Ingcaciso: Iingcali ze-pathologists zihlalutya iziphumo ngokuphonononga iipateni zokugcoba. Ukuba i-antigen ekujoliswe kuyo ikhona, i-tissue iya kubonisa i-staining ethile, njengokutshintsha kombala okanye i-fluorescence, ebonisa indawo kunye nobuninzi be-antigen.

Kutheni i-Immunohistochemistry Yenziwa?

I-Immunohistochemistry sisixhobo esiguquguqukayo esinezicelo ezahlukeneyo, ngakumbi kwinkalo ye-pathology kunye nokuxilongwa. Ngezantsi zezinye zezizathu eziphambili zokuba kutheni i-IHC yenziwe:

• Ukuxilongwa koMhlaza kunye noHlelo: I-IHC idlala indima ebalulekileyo ekuxilongweni nasekuhlelweni komhlaza. Iiseli ze-tumor zihlala zivakalisa iimpawu ezithile ezinokunceda ukuchonga uhlobo lomhlaza kunye nemvelaphi yawo. Ngokomzekelo, umhlaza webele unokuhlelwa ngokusekelwe kwintetho yamanqaku afana ne-estrogen receptors (ER), i-progesterone receptors (PR), kunye ne-HER2. I-IHC inokuchonga aba bamakishi, inceda ukumisela iindlela zonyango ezisebenzayo.
• Ukuchonga imvelaphi yethumba: I-IHC isetyenziselwa ukumisela imvelaphi yethumba xa isampulu yethishu ingabonakali lula. Ngokomzekelo, ukuba umhlaza ufunyenwe kwi-lymph node, i-IHC inokunceda ukuchonga ukuba ivela kwibele, ikholoni, okanye imiphunga ngokufumanisa iimpawu ezithile zezicubu.
• Umhlaza weSigaba kunye noQoqosho lokuQikelela: I-IHC isetyenziselwa ukuvavanya ubungakanani bohlaselo lomhlaza kwizicubu, inceda ukumisela inqanaba lomhlaza kunye ne-prognosis. Isenokusetyenziswa ukuqikelela ukuba umhlaza unokuba ndlongondlongo kangakanani na ngokusekwe kwintetho yeeproteni ezithile.
• I-Autoimmune kunye neZifo ezosulelayo: I-IHC iphinda iqeshwe ukuxilonga izifo ezizimelayo, ezifana ne-lupus, okanye ukuchonga usulelo olubangelwa yibhaktheriya okanye iintsholongwane. Iiseli ezithile zokhuselo lomzimba okanye ii-pathogens zinokuchongwa kusetyenziswa iimpawu ezithile kwiisampulu zethishu.
• Iinjongo zoPhando: Kuphando, i-IHC isetyenziselwa ukufunda ukubonakaliswa kweeprotheyini kwiiseli kunye nezicubu. Oku kunokunceda abaphandi baqonde indlela izifo ezikhula ngayo kunye nendlela iiprotheni ezahlukeneyo ezisebenzisana ngayo kwiinkqubo zesifo. Ngokuqhelekileyo isetyenziselwa ukuphuhliswa kweziyobisi kunye nokuvavanya ukusebenza kokungenelela kwonyango.

Indlela yokulungiselela i-Immunohistochemistry

I-Immunohistochemistry ngokwayo ayifuni ukulungiswa kwesigulane kuba luvavanyo lokuxilonga olwenziwa kwiisampuli zezicubu. Nangona kunjalo, la manyathelo alandelayo anokubandakanyeka ekulungiseleleni uvavanyo lwe-IHC:

• Ingqokelela yeSampuli yezicubu: Uvavanyo lwe-IHC lufuna isampulu yethishu, edla ngokufunyanwa nge-biopsy okanye ngotyando. Ukuba i-biopsy iyafuneka, umboneleli wakho wezempilo uya kuchaza inkqubo, enokuthi ibandakanye i-anesthesia yendawo, i-sedation, okanye i-anesthesia jikelele ngokuxhomekeke kwindawo kunye nobukhulu besampulu yethishu.
• Imbali yesigulana: Ukubonelela umboneleli wakho wezempilo ngembali yonyango eneenkcukacha kubalulekile ukutolika iziphumo. Ukuba unembali yosapho yemihlaza ethile okanye izifo ezizimelayo, ukwabelana ngolu lwazi kunceda iingcali zezifo zikhethe izilwa-buhlungu ezifanelekileyo kuvavanyo.
• Uphononongo lweempawu: Ngaphambi kokuba wenze nayiphi na i-biopsy okanye inkqubo yotyando eya kubonelela ngesampulu ye-IHC, ugqirha wakho unokucela uphononongo olunzulu lweempawu zakho ukukhokela ukhetho lovavanyo oluthile kunye namanqaku okuhlalutya.

Utoliko lweziphumo zovavanyo

Ukutolikwa kweziphumo ze-Immunohistochemistry kuxhomekeke kwipatheni kunye nokuqina kokungcola, okuhlalutywa yi-pathologist eqeqeshiwe. Izinto eziphambili ekufuneka ziqwalaselwe zezi:

• Ukuqina kokuqina:
  • Ibala eliqinileyo: Ibonisa umgangatho ophezulu we-antigen ekujoliswe kuyo kwi-tissue.
  • Ibala eliphakathi: Ibonisa ubukho obuphakathi kwe-antigen.
  • Ubuthathaka okanye Akukho Namabala: Iphakamisa ukuba i-antigen ayikho okanye ikhona kumanqanaba aphantsi.
• Indawo yokulahla:
  • Ukungcola kweCytoplasmic: Ukuba ibala lifunyenwe kwi-cytoplasm yeseli, iphakamisa ukuba iprotheni ekujoliswe kuyo ikhona.
  • Ukungcoliswa kweNyukliya: Ukuba ibala likwinucleus, icebisa ukuba iproteni ekujoliswe kuyo idlala indima kwinucleus yeseli.
  • Ukudyobheka kwiMembranous: Ukuhlamba kwi-membrane yeseli kunokubonisa umsebenzi we-receptor okanye i-adhesion molecules.
• Ipateni yokuNgcolisa: Ipateni yokungcolisa, kubandakanywa nokuba isasazeka okanye igxininise, inceda ukuvavanya ukusabalalisa kunye nobuninzi be-antigen kwaye isetyenziselwa ukuxilongwa kweentlobo ezithile zomhlaza okanye izifo.
• Iimpawu ezithile: Izifo ezahlukeneyo zidibene nabamakishi abathile. Umzekelo, i-HER2 positivity ibalulekile ekumiseleni iindlela zonyango kumhlaza webele, ngelixa ii-CD markers zinceda ekuhleleni iintlobo ezahlukeneyo ze-lymphoma.

Uluhlu oluqhelekileyo lwe-Immunohistochemistry

Ekubeni i-Immunohistochemistry bubuchule obusetyenziselwa ukufumanisa ii-antigens ezithile, akukho "luhlu oluqhelekileyo" lwemveli kwiziphumo zovavanyo. Kunoko, iziphumo zitolikwa ngokusekelwe ekubeni i-antigen ekujoliswe kuyo ikhona kwisampuli kunye nomzekelo wokuvakalisa. Isiphumo esihle somakishi othile sinokubonisa ubukho bomhlaza okanye isifo, ngelixa isiphumo esibi singabonisa ukungabikho kwayo. Ukutolika kuxhomekeke kwimeko yeklinikhi kunye namanqaku athile avavanywayo.

Ukusetyenziswa kwe-Immunohistochemistry

I-Immunohistochemistry inosetyenziso olubanzi kuxilongo lwezonyango, kubandakanya:

• Ukuxilongwa koMhlaza kunye nokuChwetheza kancinci: I-IHC inceda ukuchonga uhlobo lomhlaza, ukumisela imvelaphi yawo, kwaye ihlele i-subtype yayo (umzekelo, umhlaza webele, i-lymphoma).
• Ukuchonga iimpawu zeTumor: Isetyenziselwa ukubona ubukho beeprotheni ezifana ne-HER2 kumhlaza webele okanye i-CD20 kwiintlobo ezithile ze-lymphoma.
• Uxilongo lweSifo somzimba: I-IHC inokunceda ekuboneni ubukho beeseli ezithile zokuzivikela okanye ii-antibodies ezibonisa izifo ezizimelayo ezifana ne-lupus okanye i-rheumatoid arthritis.
• Uxilongo lweSifo esosulelayo: I-IHC isetyenziselwa ukuchonga usulelo kwiithishu, kubandakanya iintsholongwane, iintsholongwane, okanye usulelo lweparasitic.
• Uphando: Iqhele ukusetyenziswa kuphando ukufunda ukubonakaliswa kweprotheyini kwizicubu kunye neeseli ukuqonda iindlela zesifo, ngakumbi kuphuhliso lwamachiza.
• Umhlaza weqonga: I-IHC inokuncedisa ekumiseleni inqanaba lomhlaza, ngakumbi kwiimeko apho ukuchonga ubungakanani bokusasazeka kwesifo kubalulekile.

I-10 yemibuzo ebuzwa rhoqo malunga ne-Immunohistochemistry

1. Yintoni i-Immunohistochemistry? I-Immunohistochemistry yindlela yaselabhoratri esebenzisa izilwa-buhlungu zokubona ii-antigens ezithile kumacandelo ezicubu. Isetyenziselwa ukuxilonga izifo, ngakumbi umhlaza, kunye nokuchonga ubukho beeprotheyini ezithile kwiiseli.
2. Yenziwa njani i-Immunohistochemistry? Iisampulu zezicubu ziyaqokelelwa, ngokuqhelekileyo nge-biopsy. Isampulu ke iphathwa ngezilwa-buhlungu ezikhethekileyo kwiiproteni ezivavanywayo. Ukuba i-antigen ikhona, ibophelela kwi-antibody kwaye ivelise uphawu olubonakalayo, oluhlalutywa phantsi kwe-microscope.
3. Ziziphi iimeko ezinokuxilonga i-Immunohistochemistry? I-Immunohistochemistry isetyenziselwa ukuxilonga i-cancer (efana nebele, imiphunga, okanye i-lymphoma), izifo ezizimele, kunye nosulelo. Inceda ukufumanisa ubukho kunye nohlobo lwee-antigens ezinokubonisa isifo.
4. Yintoni indima ye-Immunohistochemistry ekuxilongweni komhlaza? I-IHC inceda ukuchonga iimpawu ze-tumor ezifana ne-HER2, i-estrogen receptors, kunye ne-progesterone receptors, ezibalulekileyo ekuxilongeni iintlobo zomhlaza, ukucofa, kunye nokuchonga unyango olufanelekileyo.
5. Ngaba i-Immunohistochemistry invasive? I-Immunohistochemistry ngokwayo ayihlaseli. Nangona kunjalo, ifuna isampulu yethishu efunyenwe ngenkqubo efana ne-biopsy okanye utyando, olunokuthi lubandakanye ukungonwabi ngokuxhomekeke kwinkqubo.
6. Kuthatha ixesha elingakanani ukufumana iziphumo kwi-Immunohistochemistry? Iziphumo ze-IHC zihlala zithatha iintsuku ezimbalwa ukuya kwiveki, kuxhomekeke kubunzima bovavanyo kunye nesidingo sohlalutyo olongezelelweyo.
7. Sithetha ntoni isiphumo esihle se-Immunohistochemistry? Isiphumo esihle sibonisa ukuba i-antigen ekujoliswe kuyo ifunyenwe kwisampuli yezicubu. Kuxilongo lomhlaza, oku kunokuthetha ubukho bophawu lomhlaza, ngelixa kwizifo ezizimelayo, kunokucebisa ubukho beeseli ezithile zokhuselo lomzimba.
8. Isiphumo esibi se-Immunohistochemistry sithetha ntoni? Isiphumo esibi sithetha ukuba i-antigen ayizange ibonwe kwisampuli yezicubu. Ekuxilongweni komhlaza, oku kunokucebisa ukuba isiphawuli esithile, esifana ne-HER2 kumhlaza wamabele, asikho.
9. Ngaba i-Immunohistochemistry ingasetyenziselwa ukwenza umhlaza? Ewe, i-IHC inceda ekwenziweni komhlaza ngokuchonga ubungakanani bokukhula kwethumba, ukusasazeka, kunye nokubonakaliswa kwamanqaku athile aphembelela ukuxilongwa kunye nezigqibo zonyango.
10. I-Immunohistochemistry yahluke njani kwezinye iimvavanyo? Ngokungafaniyo novavanyo lwemfuza okanye ukuxilongwa kweemolekyuli, i-Immunohistochemistry igxile kwinkcazo yeprotheyini kwiisampulu zezicubu, iyenza ibe luncedo kakhulu ekuboneni nasekuxilongweni kweemeko kwinqanaba leselula.

isiphelo

I-Immunohistochemistry sisixhobo esibalulekileyo sokuxilonga kumayeza anamhlanje, esinika ulwazi oluxabisekileyo malunga nobukho kunye nohlobo lwezifo, ngakumbi i-cancer kunye neemeko ze-autoimmune. Ngokufumanisa ii-antigens ezithile, inceda ii-pathologists zixilonge ngokuchanekileyo iimeko, umhlaza weqonga, kunye nokumisela eyona ndlela yonyango. Ngelixa inkqubo ayihlaseli, ukuqokelela isampulu yethishu kunokufuna ukulungiswa ngononophelo kunye nobuchule bonyango.

Ukuqonda inkqubo ye-Immunohistochemistry, ukusetyenziswa kwayo, kunye nendlela yokutolika iziphumo kunokukhokela zombini izigulane kunye nababoneleli bezempilo ekwenzeni izigqibo ezinolwazi malunga nokuxilongwa kunye nonyango. Ingaba wenza uvavanyo lwe-IHC lokuxilongwa komhlaza okanye njengenxalenye yophando lophando, le ndlela isisiseko soxilongo lwezonyango kunye nophando oluqhubekayo.