I-Paraformaldehyde iyifomu ye-polymerized ye-formaldehyde evame ukusetshenziswa ekwenzeni izinto zibe ziqinile ezinhlelweni ze-biological nezokhemikhali. Ibalulekile ngenxa yokukwazi kwayo ukukhulula igesi ye-formaldehyde uma ishisa noma ihlukaniswa, okusebenza njenge-agent yokuxhumanisa ephumelelayo. Le mpahla ivumela i-paraformaldehyde ukuba iqinise izakhiwo ze-cellular ngokwakha ama-bond e-covalent phakathi kwamaprotheni, ama-nucleic acids, nezinye izakhi zomzimba. Le ngxube ivame ukutholakala njengepudingi emhlophe ethile, lapho ihlukaniswa emanzini, ikhiqiza isixazululo se-formaldehyde esisebenza kakhulu ku-histology, immunohistochemistry, kanye ne-biology ye-molecular. Indlela yayo ehlukile ibandakanya ukungena ezicubu noma kumaseli ukuze iqinise i-morphology futhi igcine i-antigenicity, okubalulekile ukuze kwenziwe ukuhlaziywa kwe-microscopic okulandelayo nezinhlelo zokufaka imibala.

Ngenxa yokuhlanzeka kwayo okuhle kakhulu uma kuqhathaniswa nezixazululo ze-formaldehyde ezithengiswayo, i-paraformaldehyde inikeza izimo zokufaka ezilawulwayo kakhulu. Lokhu kuvumela abacwaningi ukuthi bafinyelele emiphumeleni efanele yokufunda i-morphology ye-cellular kanye nokuhlelwa kwe-protein. Ukuzinza kwekhemikhali nokulula kokulungiselela kwenza i-paraformaldehyde ibe ukukhetha okuthandwayo kokufaka ezikhungweni zokucwaninga emhlabeni jikelele. Futhi, ayivamile ukuhlinzeka futhi iyazinzile kakhulu ngesikhathi sokugcina uma iqhathaniswa ne-formaldehyde emoyeni, okwenza kube kuphephile ukuyisebenzisa nsuku zonke. Ukuqonda isimo sekhemikhali kanye nenqubo yokufaka ye-paraformaldehyde kubalulekile kubacwaningi abahlose ukuthuthukisa izinqubo zokulungiselela amasampula.

Ekuqiniseni kwemichilo, umsebenzi omkhulu we-paraformaldehyde ukuhlanganisa amaqembu e-amino kumaprotheni, ngempumelelo “ukuvula” izakhiwo zomolekyuli endaweni. Le nhlanganisela igcina ubuqotho bezinto zomzimba nezokwakheka kwamangqamuzana, ivimbela ukwehla kwe-enzymatic noma izinguquko zokwakheka ngemuva kokuhlola. Izinga lokuqinisa lingashintshwa ngokulawula ukugxila, izinga lokushisa, nesikhathi sokuqinisa, okwenza i-paraformaldehyde ibe nezinhlobonhlobo zokuhlola eziphilayo. Ngaphezu kwalokho, ukuqiniswa kwe-paraformaldehyde kuqinisekisa ukuhambisana nezindlela ezahlukene zokuhlobisa nezokubhalisa ezibalulekile ocwaningweni lwesimanje lwezinto zomzimba.

Ngokubanzi, i-paraformaldehyde iyisithako esiyisisekelo emkhakheni wokufaka amakhemikhali, inikeza ibhalansi yokusebenza, ukuzinza, nokulula kokusebenzisa. Iziphumo zayo nezindlela zenza kube kubalulekile ekugcineni amasampula obuchopho ukuze kuhlaziywe kahle ngezithombe ezincane nezokwenziwa kwamakhemikhali. Lezi zihloko ezilandelayo zizobheka ngokujulile izicelo zayo, izindlela zokulungiselela, izinselelo, nezindlela ezinhle zokusebenza ezinqubweni zokufaka.

I-Paraformaldehyde idlala indima ebalulekile ekuxhumaneni kwezakhi ze-cell ngokwakha amabhuloho e-methylene phakathi kwe-amino acids ku-proteins. Lokhu kuxhumanisa kuqinisa isakhiwo se-three-dimensional se-proteins, kugcine ukwakheka kwe-cellular futhi kuvumele izifundo ezinemininingwane ye-morphological. Ngokwehlukile kumasampula e-formaldehyde athengiswayo angase abe nezinhlaka eziphukile futhi ahluke ekugxileni, i-paraformaldehyde inikeza umthombo ohlanzekile futhi ohloniphekile we-formaldehyde. Le consistency ibalulekile ukuze kuqinisekiswe ukuphindaphindwa kokuhlola okwethembekile.

Izixazululo ze-formaldehyde ezisemthethweni zivame ukwehla ngokuhamba kwesikhathi futhi zingase zibe ne-methanol noma ezinye izithasiselo ezingenza kube nezinkinga ekubambeni. Ngokuphambene, i-paraformaldehyde, uma ilungiswe muva, ikhipha i-formaldehyde ehlanzekile enezinto ezingcolile ezincane, yehlisa ubungozi beziphumo ezimbi ekuphumeleleni kwesampula. Lokhu kuholela ekugcinweni kwe-antigen okuthuthukisiwe kanye nokuhambisana okungcono nezindlela zokumaka i-immunostaining. Kubacwaningi abafunda ukuhlela kwe-protein noma umsebenzi we-enzyme, ukubamba kwe-paraformaldehyde kuqinisekisa ukuthi izinjongo zomolekyuli zihlala ziphila futhi zifinyeleleka ukuze zixhume ne-antibody.

Indima ye-paraformaldehyde idlulela ngaphandle kokufaka kuphela; iphinde yehlise ukungcoliswa kwangemuva futhi ithuthukise amazinga we-signali-to-noise ekuhlaziyweni kwe-immunohistochemical. Ngokulawula ngokucophelela izimo zokufaka, abacwaningi bangakwazi ukuthuthukisa ibhalansi phakathi kokugcina imininingwane yeseli nokugcina ukufinyelela kumantshontsho. Le mfuneko ibalulekile ikakhulukazi ezicini eziyinkimbinkimbi lapho ukungena kokufaka kanye namantshontsho kungaba yizici ezilinganayo.

Ukubaluleka kwe-paraformaldehyde ocwaningweni kugcizelelwa kakhulu ngezicelo zayo ku-electron microscopy, lapho isebenza njengokulungisa kokuqala ngaphambi kokufaka i-glutaraldehyde. Le nhlanganisela ifinyelela ukugcina okuhle kwe-ultrastructural ngenkathi igcina i-antigenicity yokuhlonza i-immunolabeling. Ngakho-ke, i-paraformaldehyde ihlala iyithuluzi elingabalulekile kokubili ezinqubweni ezijwayelekile nezithuthukisiwe ze-biology yeseli.

Ukufaka kahle kwekhemikhali nge-paraformaldehyde kudinga ukulungiselela ngokucophelela ukuze kuqinisekiswe ukugcina kahle izakhiwo zamangqamuzana. Inqubo iqala ngokulungisa ama-slides e-microscope ahlanzekile, alungiswe kahle uma usebenza nezicucu zethishu noma amasiko ezicubu. Ama-slides kufanele abe mahhala ezitholakalayo ukuze kugwenywe ukuphazamiseka kokufakwa nokuhlobisa.

Ukulungiselela i-paraformaldehyde fixative, uthuli luyaxutshwa esixazululweni esinokubufakazi, ngokuvamile i-phosphate-buffered saline (PBS), ngokushisa kancane kuze kube sekupheleni. I-concentration evamile yokusebenza ibanga ukusuka ku-3% kuya ku-4%, kodwa lokhu kungashintshwa kuye ngohlobo lwezicubu nezinhloso zokuhlola. Kubalulekile ukulungiselela izixazululo ze-paraformaldehyde ezisha ukuze kugcinwe ukusebenza kahle kokufaka futhi kugwenywe imikhiqizo yokwehliswa.

Inqubo yokubopha ihilela ukufaka isampula esixazululweni se-paraformaldehyde ekamelweni lokushisa noma ku-4°C isikhathi esichaziwe, ngokuvamile phakathi kwemizuzu eyi-10 kuya kweyi-30. Ukulungiswa kokushisa nesikhathi kungasiza ekuthuthukiseni ikhwalithi yokubopha ngokuya ngosayizi wesampula kanye nokugcina i-antigen efunwa. Ngemuva kokubopha, izibonelo zihlanzwa ngensiza yokuhlanza ukuze kukhishwe i-fixative esele futhi kuvinjwe ukweqisa, okungase kubhujise izindawo ze-antigen.

Ukuphenywa kulandela ukujolisa ukuze kuvumele ama-antibody ukufinyelela ezinjengweni zangaphakathi. Ama-detergent afana ne-Triton X-100 noma i-saponin ajwayelekile asetshenziswa ngezinga eliphansi ngezikhathi ezimfushane ukuze kuphenywe imikhawulo yamaseli ngaphandle kokuphazamisa kakhulu ubunjalo. Lesi sigaba sibalulekile kumaphrothokholi wokumaka i-immuno futhi kumele siqinisekiswe kahle ngokuhambisana nezimo zokujolisa.

Ukulandela ngokucophelela lezi zinyathelo kuqinisekisa ukuphindaphindwa kanye nemiphumela yokubopha yekhwalithi ephezulu. Ukuze uthole imininingwane ethile mayelana nezincazelo zomkhiqizo kanye nezimpahla zamakhemikhali, abakhiqizi abafana noHebei Jintaida Chemical Co., Ltd.hlinzeka ngemikhiqizo ye-paraformaldehyde enobukhulu obuphezulu efanele ukusetshenziswa kwelebhu, ixhasa izinqubo zokulungiselela amasampula ezisebenzayo.

Naphezu kokusetshenziswa kwayo kabanzi, ukuvikelwa kwe-paraformaldehyde kuveza izinselelo eziningi okufanele zixazululwe abacwaningi ukuze bafeze imiphumela ephumelelayo. Enye yezinselelo eziyinhloko ukuqinisekisa ukufinyelela okwanele kwezithako zomzimba kumagciwane aqondiwe ngemuva kokuvikelwa. Ukuvikelwa ngokweqile kungaholela ekuxhumaneni okweqile okumboza ama-epitope, kwehlisa ukusebenza kahle kokubopha kwezithako zomzimba. Ngakolunye uhlangothi, ukuvikelwa okunganele kungaholela ekugcinweni okuphambene kwemiphumela kanye noise ephezulu.

Ukulungisa umthamo we-paraformaldehyde kanye nesikhathi sokufaka kubalulekile ekuwazini lezi zici. Abacwaningi bavame ukwenza izivivinyo zokuqala ukuze bathole izimfuneko ezinhle kakhulu zezimfanelo zabo ezithile nezibungu. Ukusebenzisa izindlela zokubuyisela i-antigen, ezifana nokubuyiselwa kwe-epitope okwenziwa ukushisa noma ukuhluza nge-enzymatic, kungathuthukisa ukufinyelela kwe-antibody ezindaweni ezibekwe.

Enye inkinga evamile ubukhulu bokungena kwe-paraformaldehyde ezicini ezinzima. Ukungena okunganele kuholela ekwakhiweni okungalingani, lapho izindawo ezisemaphethelweni ziqinile kahle kanti izindawo zangaphakathi zikhubazekile. Ukufakwa ezingeni lokushisa eliphansi isikhathi eside noma ukusebenzisa izingcezu zethishu ezincane kungasiza ekunciphiseni le nkinga.

Ukuphathwa kwezixazululo ze-paraformaldehyde kudinga izinyathelo zokuphepha ngenxa yokuba yingozi nokuphuma kwe-formaldehyde. Ukuhlinzekwa komoya okufanele, ukusebenzisa imishini yokuvikela yomuntu, nokulandela imihlahlandlela yokugcina kubalulekile ukuze kuncishiswe ubungozi bokuthintwa. Izinkinga zokuzinza zivele ngoba izixazululo ze-paraformaldehyde zingase zibe ne-depolymerization ngokuhamba kwesikhathi, kunciphisa ukusebenza kokufaka; ngakho-ke, izixazululo ezilungiselelwe muva zicebiswa ukuze kutholakale imiphumela engcono.

Ukubhekana nalezi zinkinga kuqinisekisa ukuthi i-paraformaldehyde ihlala iyisixazululo esithembekile, ivumela imiphumela yokuhlola i-immunostaining ne-microscopy ewusizo.

Ukuze uthole izinzuzo ezinkulu ze-paraformaldehyde ekugcineni kwekemikhono, kubalulekile ukulandela izinqubo ezinhle. Kuhle ukuhlela izixazululo ezintsha ze-paraformaldehyde ngokushesha ngaphambi kokusetshenziswa ukuze kuqinisekiswe ukukhululwa okuphezulu kwe-formaldehyde kanye namandla okugcina. Izixazululo kufanele zihlolwe ukuze kususwe ama-particulates futhi zigcinwe ku-4°C uma ukugcina okwesikhashana kudingeka.

Ukukhetha umthamo ofanele wokubopha, ngokuvamile phakathi kuka-3% no-4%, kuhlanganisa ukuxhumana okusebenzayo ngaphandle kokwephula ukufinyelela kwe-antigen ngokweqile. Isikhathi sokubopha kanye nezinga lokushisa kufanele kulungiswe uhlobo lwesampula, ngokubopha okufushane kwezinhlobo ezibucayi kanye nesikhathi eside kwezicubu ezinzima. Ukufaka isinyathelo sokuphumelela ngama-detergents alula ngemuva kokubopha kuthuthukisa ukufinyelela kwe-antibody ngaphandle kokulimaza izakhiwo ze-cellular.

Abasebenzisi kufanele babe nekhono mayelana nokulimala okungenzeka kwezibonelo okubangelwa ukujolisa ngokweqile nokusetshenziswa kwe-detergent ngokweqile, okungashintsha i-morphology yamaseli noma kuholele ekulahlekelweni kwezimolekyuli ezihlosiwe. Ukuhlola izinqubo ezahlukene zokujolisa kanye nezinga le-antibody kusiza ekuthuthukiseni ikhwalithi yokuhlobisa. Ngaphezu kwalokho, ukuhlanganisa ukujolisa kwe-paraformaldehyde nezinye izixazululo zokujolisa ezifana ne-glutaraldehyde kungathuthukisa ukugcina kahle kwe-ultrastructure ye-electron microscopy.

For laboratories seeking reliable chemical raw materials, companies such as Hebei Jintaida Chemical Co., Ltd.nikeza i-parafomaldehyde ephezulu kakhulu kanye nemikhiqizo ehlobene, iqinisekisa ikhwalithi ehambisanayo esekela ukuphindaphindwa kokuhlola nokuhambisana nezokuphepha.

Ukuqhubeka kwemfundo ngezindlela zokubopha nokulandela izinqubo eziphakanyisiwe kukhuthaza abacwaningi ukuba basebenzise amandla aphelele we-paraformaldehyde ekugcineni imininingwane ye-cellular kanye ne-molecular ukuze kwenziwe ucwaningo olunhlobonhlobo lwezesayensi.

Ukufakwa kwe-paraformaldehyde kuyisisekelo sokuphumelela kokumaka kwemvelo, kodwa ukuhlela izinqubo zokufaka nokumaka kungathuthukisa kakhulu ukusebenza kwe-antibody nokucaca kwesignali. Enye indlela esebenzayo ukuhlela izimo zokufaka ngokuya ngohlobo lwe-antibody kanye ne-antigen eqondiwe. Ezinye izindawo ezithile zihluleka ekuxhumaneni futhi zidinga ukufakwa okuncane noma isikhathi sokub exposure esifushane.

Ukusebenzisa izindlela zokubuyisela i-antigen ngemva kokufakwa kwe-paraformaldehyde kungavula ama-epitope futhi kwandise ukuxhumana kwe-antibody. Ukubuyiselwa kwe-epitope okuholelwa ukushisa kusetshenziswa ama-citrate noma ama-EDTA buffers kusekelwe kakhulu ukuze kubuyiselwe i-antigenicity evinjiwe ukuhlanganiswa. Ukubuyiselwa kwe-enzymatic nge-proteases kungasetshenziswa ngokucophelela ukuze kuthuthukiswe ukujolisa ngaphandle kokwephula ubuqotho bezicubu.

Ukusebenzisa i-parafomaldehyde ephezulu, ehlanzekile evela kubahlinzeki abathembekile kuthuthukisa ukujolisa okuhambisanayo futhi kunciphisa ukungafani emiphumeleni yokuhlola. Ngaphezu kwalokho, ukuhlanganisa ukujolisa kwe-parafomaldehyde nezithako zokuvimba ezifanele kusiza ukunciphisa ukubopha okungathandeki kwe-antibody kanye ne-fluorescence engemuva, ngaleyo ndlela kuthuthukisa ubudlelwano phakathi kwesignali nokukhala.

Ukulungisa izinqubo zokungena kwe-permeabilization ngama-detergents afana ne-Triton X-100 ezingeni elilungisiwe kuvumela ama-antibodies ukuba angene eziceleni futhi afinyelele ezinjengweni zangaphakathi kahle. Ngaphezu kwalokho, ukuhlela kahle ukujova nokungena kwe-permeabilization kubalulekile ukuze kugcinwe ubunjalo bezicubu ngenkathi kufinyelelwa ekubhaleni okuqinile.

Abacwaningi abanentshisekelo ku-paraformaldehyde ephezulu kanye nezinsiza zamakhemikhali bangahlola imikhiqizo etholakalayo kuHebei Jintaida Chemical Co., Ltd., umkhiqizi othembekile ogxile ekucaciseni kwemikhiqizo nasekulawuleni ikhwalithi, okubalulekile ezinhlelweni zokufaka izithombe ze-immunostaining eziphumelelayo.

I-Paraformaldehyde iseyingxenye ebalulekile yokugcinwa kwemichilo emkhakheni wezocwaningo lwezobuchwepheshe nezemvelo ngenxa yezici zayo ezihlukile kanye nezindlela ezivumela ukuhlanganiswa okuphumelelayo nokugcinwa kwezakhiwo zamangqamuzana. Ikhono layo lokukhiqiza izixazululo ezihlanzekile ze-formaldehyde lapho ihlukaniswa lenza ibe ngcono kuneziningi zezimikhiqizo ze-formaldehyde ezithengiswayo, iqinisekisa ikhwalithi yokugcinwa efanele kanye nokuphindaphinda okuqinisekisiwe kokuhlola.

Ukuqonda izindlela zokulungiselela, izinqubo zokubopha, kanye nezinselelo ezihambisana ne-paraformaldehyde kuvumela abacwaningi ukuba baphucule ukugcina amasampula ukuze kube nokuhlola kwe-microscopy nokumaka kwe-immunostaining. Ngokulandela izinqubo ezinhle kakhulu nokusebenzisa izinto ezisezingeni eliphezulu ezivela kubahlinzeki abathembekile njengeHebei Jintaida Chemical Co., Ltd., abasebenzisi bangathola imiphumela engcono yokubopha ethuthukisa ukuhlaziywa okunembile kwezicubu nemolekyuli.

Ukuthuthuka okuqhubekayo kwezindlela zokubopha kanye nemfundo mayelana nezici ze-paraformaldehyde kuqhubeka nokuthuthukisa ukusetshenziswa kwayo emikhakheni ehlukene yesayensi, kusukela ku-histology kuya ku-molecular biology. Ekugcineni, ukubaluleka kwe-paraformaldehyde kusemkhakheni wayo wokugcina ubunjiniyela obuyinkimbinkimbi bezibonelo zokuphila, kuvumela abacwaningi ukuba bahlolisise ubunzima bokuphila ngokuqondile nangokucacile.