Yintoni i-Immunostaining? Masithethe ngolu vavanyo lokufumanisa izifo ngokulula!

Ngaba wakha wacelwa ngugqirha ukuba akwenze uvavanyo olubizwa ngokuba yi-"Immunostaining"? Okanye wakha waliva eli gama ndaweni ithile? Mhlawumbi livakala linzima kuwe. Kodwa ungakhathazeki, namhlanje siza kuthetha ngolu vavanyo olubizwa ngokuba yi-Immunostaining, into oluyenzayo, indlela oluqhutywa ngayo, yonke into ngendlela elula kakhulu onokuyiqonda. Njengokuthetha nomhlobo.

Yintoni kanye kanye i-immunostaining?

Ngamafutshane, i-immunostaining luvavanyo olukhethekileyo olusetyenziswa ziingcali zethu zezifo. Isetyenziselwa ukuchonga imolekyuli ethile yeproteni phakathi kweeproteni ezininzi kwisampulu ethathwe emzimbeni wakho, njengegazi, umongo wethambo, okanye izicubu. Cinga ngayo njengokufumana umntu othile kwisihlwele. Le ndlela iluncedo kakhulu ekuchongeni izifo ezahlukeneyo, ngakumbi usulelo kunye nomhlaza.

Lusebenza njani olu vavanyo?

Kulungile, ngoku masibone indlela esebenza ngayo le immunostaining. Isebenzisa into ebizwa ngokuba yi-antibodies . La ma-antibodies afana namajoni kwinkqubo yethu yokuzikhusela yomzimba. Ayaqaphela kwaye anamathele kwizinto ezibangela izifo, ezibizwa ngokuba yi-antigens . Ngoko ke, kolu vavanyo, oogqirha basebenzisa i-antibody ekhethekileyo kuloo protein (ebizwa ngokuba yi-antigen) abayifunayo.

Emva koko kuza inxalenye "yokungcolisa". Kufana nokugqamisa isivakalisi esibalulekileyo encwadini. Kodwa akulula njengokufaka umbala kwi-T-shirt. Ezi ndlela "zokufaka umbala" zisebenzisa iidayi ezikhethekileyo, ii-enzymes ezitshintsha umbala, iimolekyuli ze -fluorescent , kwaneenxalenye zegolide. Ezi zilebhelisha iiproteni kunye neeseli abazifunayo, ukuze abaphandi bazibone ngokucacileyo phantsi kwe-microscope.

Ngaphambi kokuba enze olu vavanyo lokufaka i-immunostaining, ugqirha uza kuthatha isampuli yegazi lakho, umongo wethambo, okanye izicubu zomzimba. Emva koko iya kuthunyelwa kwingcali yezifo ukuze ivavanywe. Khumbula, nokuba isampuli ithathwa esibhedlele okanye eofisini kagqirha, olu vavanyo lokufaka i-immunostaining luhlala lwenziwa kwilabhoratri, okanye oko sikubiza ngokuba yilabhoratri.

Ziziphi iintlobo ze-immunostaining?

Kukho iindlela ezahlukeneyo zokukhusela amajoni omzimba ezisetyenziswa ngoogqirha. Indlela nganye inenkqubo ethile yovavanyo, okanye iprotokholi, enxulumene nayo. Nazi ezinye zeendlela eziphambili ezisetyenziswayo:

• I-Immunohistochemistry
• I-flow cytometry
• I-immuno-electron microscopy (i-EM immunolabeling)

Ngaba masijonge nganye kwezi nkcukacha ngeenkcukacha ezithe vetshe?

I-Immunohistochemistry - indlela esetyenziswa kakhulu

Le yindlela esetyenziswa kakhulu yokujonga i-immunostaining . Kwi-immunohistochemistry, iqhekeza elincinci lesampuli libekwa kwisilayidi seglasi, kwaye i-antibody ebopha kwindawo ekujoliswe kuyo yongezwa kuyo. Emva koko idaywa ngeedayi ezikhethekileyo okanye ii-enzymes. Ezi zibizwa ngokuba zii-immunostains. Oku kuvumela ingcali yezifo ukuba ibone iiproteni ezithile kunye neeseli ngokucacileyo phantsi kwe-microscope.

Jonga ukuba zeziphi izibonelelo ze-immunohistochemistry:

• Inceda ekuchongeni iintlobo ezahlukeneyo zomhlaza.
• Inceda ekuhleleni iimeko ezininzi zemithambo-luvo kunye neengxaki zegazi.
• Oku kukwavumela oogqirha ukuba bafunde ngemiphumo, okanye indlela ezibonakala ngayo ii-genes ezithile.

I-Flow cytometry - njengomatshini obala kwaye wahlula iiseli

Kwindlela ebizwa ngokuba yi-flow cytometry, ingcali isebenzisa ii-antibodies ezineelebheli ezinemibala ukuphawula iiproteni ezizifunayo. Emva koko, kusetyenziswa umqadi welaser okhethekileyo ukubala, ukuhlela, nokwahlula iiseli zakho nganye nganye. Kufana nokuskena ukutya kwivenkile enkulu.

Oogqirha basebenzisa i-flow cytometry ukuxilonga izifo ezifana nezi:

• Izifo ezosulelayo
• Izifo ezizikhusela ngokwazo emzimbeni
• I-Leukemia
• I-Lymphoma

I-immuno-electron microscopy (i-EM immunolabeling)

I-immuno-electron microscopy ivumela ingcali yezifo ukuba ibone isampuli yakho ngokwandiswa okukhulu kakhulu nangeenkcukacha ezicacileyo. Kule nto, iingcali zezifo zincamathisela amasuntswana egolide amancinci abizwa ngokuba ngamasuntswana egolide e-colloidal kwi-antibodies. Le golide ye-colloidal kulula kakhulu ukuyibona phantsi kwe-electron microscope. Ngoko ke, xa loo antibody ihamba ize ibambe i-antigen, amasuntswana egolide ayenza ibonakale ngokucacileyo njengamachaphaza amnyama.

Ukongeza ekufumaneni izifo ezahlukeneyo zentsholongwane, i-immuno-electron microscopy inokwenzaNditsho ndifumane ii-antigens zentsholongwane kwiigonyo.

Ndingazilungiselela njani ezi vavanyo?

Ngokwesiqhelo, akufuneki ulungiselele uvavanyo lwe-immunostaining. Nangona kunjalo, ugqirha wakho uya kukunika imiyalelo ngaphambi kokuba uthathe isampuli yegazi lakho, yezicubu, okanye yomongo wethambo. Oku kuya kukunceda wazi kakuhle into omawuyilindele. Kubalulekile ukumamela ngononophelo oko kuthethwa ngugqirha wakho.

Uvavanyo luthatha ixesha elingakanani? Kuthatha ixesha elingakanani ukuba iziphumo zibuye?

Oku kuxhomekeke kakhulu kwindlela yokugonya amajoni omzimba ayisebenzisayo ugqirha wakho. Ezinye iindlela zithatha iiyure ezimbalwa kuphela , ngelixa ezinye zingathatha ukuya kuthi ga kwiintsuku ezili-10 . Ugqirha wakho uya kukwazi ukukuxelela ukuba kuza kuthatha ixesha elingakanani ngokwemeko yakho.

Ixesha elithathayo ukuze iziphumo zibuye nalo liyafana. Ezinye iindlela zokufaka i-immunostaining zingakunika iziphumo ngemini nje enye. Ezinye zingathatha iveki okanye ngaphezulu. Xa usiya kunika isampuli, ugqirha wakho uya kukwazisa ukuba ungalindela nini iziphumo.

Ngoko ke, ngeli xesha linje, kungcono ungoyiki kwaye ube nomonde. Thetha nogqirha uze umchazele yonke into.

Zithini iziphumo? Kwenzeka ntoni ukuba aziqhelekanga?

Ingcali yezifo iza kuhlalutya iziphumo zovavanyo lwakho ize izithumele kugqirha wakho. Iziphumo ozifumanayo zinokwahluka ngokuxhomekeke kuhlobo lovavanyo olwenziwe yingcali yezifo. Ugqirha wakho uza kuchaza ngokweenkcukacha iziphumo zovavanyo lwakho kunye nentsingiselo yazo kwimeko yakho ethile.

Khumbula, ukuba iziphumo zovavanyo aziqhelekanga , oko kuthetha ukuba ingcali yezifo ifumana iimpawu zesifo, ungoyiki. Ugqirha wakho uza kukuchazela iziphumo aze axoxe nawe ngeendlela zonyango lwakho. Kwezinye iimeko, iqela loogqirha lingasebenzisana ukuphuhlisa isicwangciso sonyango esingcono.

Okokugqibela, ezona zinto zibalulekileyo ekufuneka uzikhumbule

Kulungile, ngoko ke ngokwezinto esizithethileyo, ndiyathemba ukuba unembono elungileyo malunga nokukhusela amajoni omzimba. Nazi izinto ezibalulekileyo ekufuneka uzikhumbule:

• Uvavanyo lwe-immunostaining luvavanyo lwelebhu olusebenzisa ii-antibodies ukufumanisa nokulebhelisha ii-antigen ezithile ngaphakathi kwiiseli zethu.
• Olu vavanyo lunokunceda ekuchongeni iimeko ezahlukeneyo zempilo, kubandakanya usulelo lwentsholongwane kunye neebhaktheriya, kunye nezinye iintlobo zomhlaza.
• Ukuba kufuneka wenze uvavanyo lokukhusela amasosha omzimba, akukho sizathu sokoyika.Ugqirha wakho uza kukuchazela ngokweenkcukacha.
• Nje ukuba iziphumo zovavanyo zifike, thetha ngokukhululekileyo nogqirha wakho malunga nokuba zithetha ukuthini ngemeko yakho kunye nokuba ngaba unyango luyafuneka okanye alufuneki.

Khumbula, isayensi yezonyango itshintsha rhoqo. Kungovavanyo olufana nolu apho sinokubona izifo ezininzi kwangethuba size sizinyange ngokufanelekileyo. Ngoko ke, ukuba ugqirha ucebisa uvavanyo olunjalo, kubalulekile ukululungiselela.

Ukukhusela amasosha omzimba , iPathology, iiAntibodies, iiAntigens, uvavanyo lomhlaza, uvavanyo lwelebhu