BLOG BLOG - Cov khoom ntawm Zaub Mov Kev Nyab Xeeb upstream & downstream supply chain
pob_logo
txhaj tshuaj tiv thaiv kab mob

Kev kuaj kab mob immuno-histochemistry yog dab tsi?

Muab lo rau  Cuaj hlis 26, 2020, Kho los ntawm Jason, Qeb  
Ballya-tshuaj tua kab mob-kuaj
ballya-tshuaj tua kab mob-kuaj

Nreeb Navigation

txhaj tshuaj tiv thaiv kab mob
txhaj tshuaj tiv thaiv kab mob

Immunohistochemistry yog dab tsi?

1. Lus txhais

Immunohistochemistry (ib ihc), kuj paub li immunocytochemistry, hais txog kev siv thev naus laus zis tshiab uas tsim nyog, hauv ib cheeb tsam, thiab txiav txim kom muaj nuj nqis ntawm qhov sib thooj antigens tau ua los ntawm cov tshuaj tiv thaiv kab mob antigen-antibody thiab cov tshuaj tiv thaiv kab mob histochemical hauv cov ntaub so ntswg 'qhov chaw tshwj xeeb tshuaj tiv thaiv sau nrog chromogenic reagent.

Nws ntse ua ke qhov tshwj xeeb ntawm kev tiv thaiv kab mob tiv thaiv kab mob thiab kev pom ntawm keeb kwm keeb kwm. Nrog kev pab los ntawm kev ntsuas thiab nthuav dav ntawm lub tshuab tsom me me (suav nrog cov tsom iav fluorescence thiab lub tshuab hluav taws xob me me), nws tuaj yeem tshawb pom ntau yam tshuaj antigenic (xws li protein, polypeptide, enzymes, hormones, pathogens thiab receptors, thiab lwm yam). Immunohistochemistry thev naus laus zis tau txhim kho sai heev nyob rau xyoo tas los no. Hauv xyoo 1950, nws tau txwv rau kev siv tshuab thev naus laus zis. Thiab txij thaum ntawd los cov cuab yeej siv tshuaj tiv thaiv kab mob tiv thaiv kab mob tau zoo dua thiab tau siv ntau zuj zus.

2. Kev ua phoojywg

Kev khi ntawm tshuaj tiv thaiv thiab antigen muaj qib siab ntawm qhov tshwj xeeb, thiab txhaj tshuaj tiv thaiv kab mob siv txoj cai no. Ua ntej tshaj plaws, qee yam tshuaj lom neeg hauv cov ntaub so ntswg lossis cell tau muab rho tawm thiab siv los ua antigen lossis hapten. Tom qab txhaj tshuaj tiv thaiv tsiaj, tau txais cov tshuaj tiv thaiv tshwj xeeb, thiab tom qab ntawd cov tshuaj tiv thaiv kab mob tau siv los txhawm rau txheeb xyuas tib yam tshuaj antigen hauv cov nqaij lossis ntawm tes. Vim tias qhov nyuaj ntawm antigen thiab tshuaj tiv thaiv tsis muaj xim, nws yog qhov tsim nyog los qhia qhov chaw sib khi ntawm antigen thiab tshuaj tiv thaiv los ntawm kev siv keeb kwm keeb kwm kom ua tiav qhov txiaj ntsig zoo, hauv ib cheeb tsam lossis kev tshawb fawb ntau ntawm cov antigens tsis paub hauv cov nqaij lossis cov cell.

3. Cov yam ntxwv

A. Muaj qhov tshwj xeeb

Lub hauv paus ntsiab lus ntawm kev tiv thaiv kab mob tiv thaiv txiav txim siab tias kev khi ntawm antigen thiab antibody yog qhov tshwj xeeb. Yog li ntawd, txhaj tshuaj tiv thaiv kab mob yog theoretically qhov kev qhia tshwj xeeb ntawm antigens hauv cov nqaij mos. Piv txwv li, keratin qhia txog cov khoom hauv epithelial, thiab LCA qhia txog lymphocytes. muaj. Tsuas yog thaum muaj cov ntoo khaub lig-antigens hauv cov nqaij mos, cov tshuaj tiv thaiv kab mob yuav tshwm sim.

B. Siab rhiab heev

Hauv thawj theem ntawm kev thov txhaj tshuaj tiv thaiv kab mob, vim yog kev txwv cov txuj ci, tsuas yog txoj hauv kev ncaj qha, tsis siv txoj hauv kev thiab lwm yam thev naus laus zis uas tsis tshua muaj txiaj ntsig tau muaj. Lub sijhawm ntawd, cov tshuaj tiv thaiv kab mob tsuas tuaj yeem diluted rau ob peb zaug lossis kaum ob zaug; tam sim no txoj kev tshwm sim ntawm ABC txoj kev lossis SP tso cai rau cov tshuaj tiv thaiv kom yaj ntau txhiab leej, ntau txhiab leej, lossis ntau pua lab lub sijhawm, thiab cov tshuaj tiv thaiv tseem tuaj yeem khi rau antigens hauv cov nqaij mos. Qhov tshuaj tiv thaiv kab mob-antigen rhiab heev ua rau immunohistochemistry cov txheej txheem ntau dua thiab yooj yim dua rau niaj hnub kuaj kab mob.

C. Qhov chaw raug, ua ke ntawm daim ntawv thiab kev ua haujlwm

Cov thev naus laus zis no tuaj yeem txheeb xyuas qhov tseeb antigens hauv cov ntaub so ntswg thiab cov cell los ntawm cov tshuaj tiv thaiv antigen-antibody thiab cov tshuaj tiv thaiv xim, yog li sib txawv antigens tuaj yeem muab tso rau thiab pom hauv tib cov ntaub so ntswg lossis cell tib lub sijhawm. Yog li kev sib xyaw ntawm morphology thiab kev ua haujlwm tuaj yeem kawm, uas muaj txiaj ntsig zoo los ua qhov kev tshawb fawb tob hauv thaj chaw ntawm kab mob.

4. Classification

  • Raws li cov hom ntawv sau tshuaj, xws li cov xim zas xim, cov tshuaj siv hluav taws xob, cov enzymes (feem ntau yog cov noob zaub peroxidase thiab alkaline phosphatase), ferritin, colloidal kub, thiab lwm yam, nws tuaj yeem faib ua txhaj tshuaj tiv thaiv kab mob, radioimmunoassay, immunoenzyme Labeling method thiab tiv thaiv kab mob kub thiab nyiaj.
  • Raws li cov txheej txheem zas xim, nws tuaj yeem faib ua txoj hauv kev ncaj qha (tseem hu ua txheej txheem ib kauj ruam) thiab txoj hauv kev tsis ncaj (ob kauj ruam, peb kauj ruam lossis ntau qib txheej txheem). Piv nrog rau txoj hauv kev ncaj, kev nkag siab ntawm txoj hauv kev ncaj ncees tau zoo dua qub.
  • Raws li txoj kev khi, nws tuaj yeem faib ua antigen-antibody khi, xws li peroxidase-antiperoxidase (PAP) txoj kev; kev sib raug zoo, xws li avidin-biotin-peroxidase complex (ABC) txoj kev, saw Mould avidin-peroxidase linkage (SP) txoj kev, ntawm cov txheej txheem SP yog txoj kev siv ntau dua; polymer txuas, xws li npaj tau-siv-ob txoj hauv kev, uas tshwj xeeb tshaj yog haum rau qib siab endogenous biotin cov ntsiab lus Tissue antigen kuaj pom.

5. muaj nuj nqi

A. qhov hnoos qeev

Cov kev sim feem ntau siv cov ntaub so ntswg thiab cov qauv ntawm tes. Tus qub suav nrog paraffin embedding ntu (kab mob pathological thiab cov nqaij nqaij) thiab ntu khov, thiab tom kawg suav nrog cov ntaub so ntswg luam tawm, cov ntawv swb ntawm tes thiab smears ntawm tes.

Ntawm lawv, ntu paraffin yog siv feem ntau thiab txheej txheem yooj yim tshaj plaws los ua cov ntaub so ntswg. Nws tau khaws cia zoo rau cov ntaub so ntswg morphology thiab tuaj yeem siv rau ntu ntu, uas yog qhov muaj txiaj ntsig zoo rau ntau yam kev tswj hwm kev soj ntsuam; nws kuj tseem tuaj yeem khaws cia ntev rau kev tshawb fawb yav tom ntej; Nws yuav muaj qee yam cuam tshuam rau kev tiv thaiv antigen, tab sis antigen rov qab los tuaj yeem ua tiav, uas yog txoj hauv kev nyiam ntawm kev npaj cov ntaub so ntswg hauv txhaj tshuaj tiv thaiv kab mob.

B. tshuaj tiv thaiv kab mob

Feem ntau siv cov tshuaj tiv thaiv kab mob hauv kev sim tshuaj tiv thaiv kab mob immunohistochemical yog monoclonal antibody thiab polyclonal tshuaj tiv thaiv kab mob. Monoclonal antibodies yog cov tshuaj tiv thaiv los ntawm B lymphocyte clone thiab tau npaj los ntawm kev tiv thaiv tsiaj nrog cell fusion hybridoma thev naus laus zis. Polyclonal antibody yog cov tshuaj tiv thaiv kab mob tau los ntawm tsiaj cov ntshav tom qab lub antigen purified tau txhaj tshuaj ncaj qha. Nws yog qhov sib xyaw ntawm cov tshuaj tiv thaiv kab mob tsim los ntawm ntau yam B lymphocyte clones.

C. Hom kev zas xim

Raws li cov cim sib txawv, nws tau muab faib ua txhaj tshuaj tiv thaiv kab mob method, immunoenzyme labeling method, thiab affinity histochemistry method. Qhov tom kawg yog txoj hauv kev tshawb nrhiav raws li cov khoom muaj kev sib raug zoo rau qee yam ntawm cov ntaub so ntswg. Txoj hauv kev no muaj kev nkag siab ntau dua thiab ua kom yooj yim rau kev nrhiav chaw ntawm cov kab mob antigens (tshuaj tiv thaiv) ntawm qib cellular lossis subcellular. Ntawm lawv, biotin-avidin staining method yog siv feem ntau.

Dab tsi yog immunohistochemistry siv rau?

Immunohistochemical staining muaj lub luag haujlwm dav heev hauv kev tshawb fawb biomedical thiab koom nrog ntau qhov kev tshawb fawb. Txawm li cas los xij, txhaj tshuaj tiv thaiv kab mob thev naus laus zis kuj muaj nws cov kev txwv. Piv txwv li, cov tshuaj ntsuas hauv cov ntaub so ntswg yuav tsum yog cov tshuaj tiv thaiv kab mob, thiab yuav tsum muaj kev saib xyuas tshwj xeeb. Cov tshuaj tiv thaiv kab mob tsis muaj zog tiv thaiv tsis tuaj yeem txiav txim siab los tsim tawm tshiab los ntawm tes lossis thauj los ntawm cov cell.

Yog li, cov yam ntxwv no yuav tsum tau txiav txim siab tag nrho hauv kev sim tsim. Yog tias qhov kev sim xav tau los ua pov thawj dab tsi ntawm cov cell uas paub cov protein tau muab coj los ua ke, molecular hauv situ hybridization nyiam siv tshuab Txhawm rau qhia cov pib siv txhaj tshuaj tiv thaiv kab mob thev naus laus zis tsim nyog thiab txawj ntse hauv kev sim tsim, cov hauv paus ntsiab lus ntawm nws daim ntawv thov tau piav qhia luv luv raws li hauv qab no:

  1. Txiav txim seb hom cell thiab morphology. Qee cov protein hauv cov nqaij mos muaj cov nqaij tshwj xeeb, xws li glial fibrillary acidic protein (GFAP) tsuas muaj nyob hauv astrocytes. Neurofilament (Neurofilament, NF) tsuas muaj nyob hauv cov leeg hlwb. Cov protein tshwj xeeb ntawm cov nqaij no feem ntau raug xa mus ua cov cim protein (Proteiniliaker). Hom cell tuaj yeem txiav txim siab los ntawm cov tshuaj tiv thaiv tshwj xeeb ntawm cov ntawv sau ua cov protein.

    Qee lub cell (xws li Langerhans cells thiab melanocytes hauv daim tawv nqaij) tsis yooj yim los txheeb xyuas hauv lub tsom iav qhov muag. Los ntawm kev ua immunohistochemical staining ntawm cov protein tshwj xeeb hauv cytoplasm, cov qauv ntawm cov hlwb no tuaj yeem pom meej. Cov txiaj ntsig no tshwj xeeb tshaj yog tsim nyog hauv kev tshawb fawb neuroscience thiab qog qog noj ntshav hauv tsev kho mob.
  2. Txheeb xyuas qhov chaw ntawm cov khoom lag luam ntawm tes. Saib qee yam khoom siv ntawm tes ua antigens, npaj cov tshuaj tiv thaiv sib xws, thiab ua haujlwm immunohistochemical staining ntawm cov ntaub so ntswg kom txiav txim siab qhov chaw ntawm cov khoom lag luam ntawm tes. Piv txwv li, feem ntau ntawm cov tshuaj hormones tsim los ntawm cov qog endocrine tuaj yeem txheeb xyuas los ntawm cov txheej txheem tiv thaiv kab mob immunohistochemical. Raws li qhov no, kev ua haujlwm zais cia ntawm cov cell thiab kev faib tawm ntawm cov qog endocrine tuaj yeem kawm, thiab cov qog uas tso cov tshuaj hormones ectopic tuaj yeem kuaj pom kom nkag siab txog qib sib txawv ntawm tes.
  3. Txiav txim siab txog qib kev sib txawv ntawm tes. Cov cell sib txawv ntawm tib hom qhia tawm cov cim sib txawv ntawm cov protein, thiab qib sib txawv ntawm cov cell tuaj yeem txiav txim siab raws li kev txheeb xyuas cov protein sib txawv no. Piv txwv li, cov cim tseem ceeb ntawm cov hlwb neuroepithelial yog nestin, uas qhia txog vimentin thaum nws sib txawv rau hauv cov kab mob radial glial. Hauv theem neurogenesis, nws qhia tawm - thaum nws sib txawv rau hauv neuroblasts neural tubulin (TUJl) thiab neurofilament (NF) thaum neuroblasts sib txawv ua cov neurons paub tab.
  4. Taug qab cov kab mob paj hlwb thiab nws thaj chaw kwv yees. Txoj kev tiv thaiv kab mob immunohistochemical feem ntau ua ke nrog txoj kev thauj khoom axoplasmic los kawm txog kev sib txuas ntawm cov neurons. Txoj kev thauj mus los ntawm axoplasmic siv qee yam tshuaj uas tuaj yeem nqa los ntawm cov hlab ntsha xaus. Siv cov txheej txheem histochemical los qhia tus qauv ntawm cov neurons. Feem ntau siv tracers suav nrog horseradish peroxidase thiab kub fluorescent.

    Piv txwv li lub hom phiaj yog los soj ntsuam qhov kev kwv yees ntawm cov hlab ntsha los ntawm cov keeb hauv cov hlab ntsha hauv nruab nrog cev lossis hauv nruab nrab lub paj hlwb. Ua ntej tshaj plaws, tus neeg taug qab tau txhaj tshuaj rau hauv cov hlab ntshav ntawm cov tsiaj kom tso cai rau tus tsiaj kom muaj sia nyob rau lub sijhawm. Cov ntaub ntawv raug coj los ntawm qhov chaw xav tias muaj cov hlab ntsha ua rau lub paj hlwb, thiab tus neeg taug qab yog nyob ntawm cov txheej txheem keeb kwm keeb kwm, thiab tom qab ntawd cov txheej txheem immunohistochemical tau siv los txiav txim siab nws qhov xwm txheej.
  5. Kev thov hauv chaw kho mob pathology. Xws li txheeb xyuas qhov xwm txheej ntawm qhov txhab, nrhiav pom qhov txhab me me, tshawb nrhiav keeb kwm lossis qhov sib txawv ntawm cov qog nqaij hlav, txiav txim siab theem ntawm cov qog, qhia kev kho mob thiab kev ua haujlwm yav dhau los, pab hauv kev kuaj mob thiab cais cov kab mob, thiab tshawb nrhiav qhov ua rau kis mob, lwm yam.

Immunohistochemitry raws tu qauv

1. Immunofluorescence txoj kev

Nws yog txheej txheem txheej txheem immunohistochemistry ntxov tshaj plaws. Nws siv lub hauv paus ntsiab lus ntawm antigen-antibody tshwj xeeb khi. Ua ntej tshaj plaws, cov tshuaj tiv thaiv kab mob uas paub zoo tau sau nrog cov tshuaj fluorescein, uas yog siv los tshuaj xyuas txhawm rau txheeb xyuas cov tshuaj tiv thaiv kab mob sib xws hauv lub cell lossis nqaij. Yog tias koj pom nws nyob hauv lub tshuab tsom iav fluorescence, nws yuav tso tawm qhov ci ci ntawm qee qhov nthwv dej thaum lub teeb fluorescein hauv cov tshuaj tiv thaiv kab mob antigen-antibody raug cuam tshuam los ntawm lub teeb pom kev zoo, nws. Ces tus localization ntawm ib yam antigen hauv cov ntaub so ntswg tuaj yeem txiav txim siab, thiab kev tshuaj xyuas kom muaj nuj nqis tuaj yeem ua tiav. Vim tias nws qhov tshwj xeeb muaj zog, kev nkag siab siab, nrawm thiab yooj yim, kev siv tshuaj tiv thaiv kab mob tau siv dav hauv kev kuaj mob thiab kuaj mob.

Daim ntawv thov

  • Kev thov hauv cov kab mob autoimmune
  • Kev txheeb xyuas tus kab mob thiab kab mob sai
  • Tshawb nrhiav thiab tshawb fawb txog kab mob parasites

2. Immunoenzyme labeling method

Cov txheej txheem sau npe immunoenzyme yog txheej txheem tsim nyob rau xyoo 1960 ua raws los ntawm txhaj tshuaj tiv thaiv kab mob. Lub hauv paus ntsiab lus yog thawj zaug siv cov enzyme-labeled antibodies los cuam tshuam nrog cov ntaub so ntswg lossis cov cell, thiab tom qab ntawd ntxiv cov txheej txheem enzyme los tsim cov khoom uas tsis muaj xim lossis cov xim uas muaj qee qhov hluav taws xob.

Los ntawm lub teeb lossis lub tshuab hluav taws xob me me, ntau hom xov tooj ntawm tes thiab cov tshuaj tiv thaiv kab mob hauv cov hlwb tau raug tshawb fawb hauv ib puag ncig. Immunoenzyme labeling tshuab yog siv tshuab ntau tshaj plaws. Qhov txiaj ntsig tseem ceeb ntawm txoj kev no piv nrog cov thev naus laus zis immunofluorescence yog: qhov chaw raug, qhov sib piv zoo, kev khaws cia ntev ntawm cov hnoos qeev, thiab tsim nyog rau kev tshawb fawb kho qhov muag thiab tshuab hluav taws xob.

Cov txheej txheem cim npe immunoenzyme tau tsim los sai heev, thiab muaj ntau txoj hauv kev sau npe. Nrog kev txhim kho txuas ntxiv mus thiab kev hloov pauv tshiab ntawm txoj kev, nws qhov tshwj xeeb thiab kev nkag siab tau zoo dua qub, yooj yim dua rau siv. ABC txoj hauv kev, SP txheej txheem peb kauj ruam, npaj kom siv tau ob kauj ruam txoj hauv kev nrhiav kom tau cov tshuab tau siv dav hauv kev kuaj mob.

Daim ntawv thov

  • Txhim kho qhov tseeb ntawm kev kuaj mob pathological
  • Kev siv tshuaj kho mob oncogene protein
  • Txheeb xyuas qhov qib ntawm cov qog nqaij hlav loj hlob
  • Kev tshawb pom ntawm micrometastases
  • Qhov tseem ceeb hauv kev tsim cov qog nqaij hlav
  • Qhia kev kho cov qog
  • Pab hauv kev kuaj mob ntawm kab mob tiv thaiv kab mob thiab kab mob sib kis

3. Kev siv tshuaj tiv thaiv kab mob colloidal kub

Kev tiv thaiv kab mob colloidal kub thev naus laus zis siv cov hlau tshwj xeeb tshwj xeeb xws li kub colloidal ua tus cim. Colloidal kub hais txog hydrosol ntawm kub, uas tuaj yeem nqus tau cov protein sai thiab ruaj khov uas tsis muaj cov txiaj ntsig pom tseeb ntawm kev ua haujlwm lom neeg ntawm protein. Yog li ntawd, siv colloidal kub-daim ntawv lo thawj antibody, Secondary antibody, lossis lwm yam molecules uas tshwj xeeb khi rau immunoglobulins (xws li staphylococcal protein A) raws li kev tshuaj xyuas, tuaj yeem nrhiav tau qhov muaj txiaj ntsig, lossis txawm tias muaj pes tsawg antigens hauv cov nqaij lossis hlwb.

Txij li cov colloidal kub muaj qhov sib txawv ntawm qhov sib txawv thiab cov hluav taws xob ntom ntom ntawm colloidal kub siab, cov txheej txheem tiv thaiv kab mob immuno-colloidal tshwj xeeb yog tsim rau ib daim ntawv lo lossis ntau daim ntawv lo hauv zos kev tshawb fawb ntawm immunoelectron tsom iav. Thiab vim tias xim ntawm colloidal kub los ntawm lub teeb liab mus rau xim liab, nws kuj yog qhov tsim nyog rau lub teeb tsom tsom iav. Xws li daim ntawv thov nyiaj-txhim kho kev tiv thaiv kab mob tiv thaiv kab mob thiab nyiaj tau yooj yim dua rau kev tsom teeb tsom tsom iav.

xaus

Vim nws tus kheej cov yam ntxwv tshwj xeeb, txhaj tshuaj tiv thaiv kab mob thev naus laus zis tau dhau los ua cov txheej txheem feem ntau xaiv los ntawm cov kws tshawb fawb feem ntau hauv cov ntaub so ntswg ntawm tes, kev tsim nyog thiab kev tshawb fawb ntau. Yog li, nrog kev hloov kho tshiab ntawm cov cuab yeej tshawb fawb tshawb fawb, cov kws tshawb fawb yuav tsum muaj kev nkag siab ntau dua thiab nkag siab tob txog ntawm txhaj tshuaj tiv thaiv kab mob, ua kom zoo dua tsim cov txheej txheem kev, thiab txhawb kev txhim kho ntxiv ntawm txhaj tshuaj tiv thaiv kab mob technology.

Xav kawm paub ntau ntxiv txog antigens thiab tshuaj tiv thaiv kab mob? Nyeem txog FC sawv ntaub ntawv.

References

Tsis ntev los no Posts

ZAUB MOV KEV RUAJ NTSEGMYCOTOXIN TESTPESTICIDE TESTHONEY TESTMILK TESTEGG TESTLIQUID HLOOV
Zoo siab tsim los ntawm BALLYA
LinkedIn facebook Pinterest youtube rss twitter instagram facebook-seem rss-seem linkedin-seem Pinterest youtube twitter instagram