鑽у厜鏄惧井闀滄妧鏈殑鍩烘湰鍘熺悊鏄€熷姪鑽у厜鍓傝缁嗚優鎴愬垎鍛堢幇楂樺害鍏蜂綋鐨勫彲瑙嗗寲鏁堟灉锛屾瘮濡傚湪鐩殑铔嬬櫧鍚庨潰杩炰竴涓€氱敤鐨勮崸鍏夎泲鐧解€擥FP銆傚湪缁勭粐鏍锋湰涓紝鐩殑鍩哄洜鏃犳硶杩涜鍏嬮殕锛屽垯闇€瑕佺敤鍏嶇柅鑽у厜鏌撹壊绛夊叾浠栨妧鏈墜娈垫潵瑙傚療鐩殑铔嬬櫧銆備负姝わ紝灏遍渶瑕佸埄鐢ㄦ姉浣擄紝杩欎簺鎶椾綋杩炴帴鍚勭涓嶅悓鐨勮崸鍏夋煋鏂欙紝鐩存帴鎴栭棿鎺ュ湴涓庣浉搴旂殑闈剁粨鏋勭浉缁撳悎銆傛澶栵紝鍊熷姪鑽у厜鏌撴枡锛岃崸鍏夋樉寰暅鎶€鏈笉鍙眬闄愪簬铔嬬櫧璐紝瀹冭繕鍙互瀵规牳閰搞€佽仛绯栫瓑鍏朵粬缁撴瀯杩涜鏌撹壊锛屽嵆渚块挋绂诲瓙绛夐潪鐢熺墿鐗╄川涔熷彲浠ユ娴嬪嚭鏉ャ€傛湰鏂囧氨瀵瑰嚑绉嶅父鐢ㄧ殑鑽у厜鍓傝繘琛屼簡鍏蜂綋鐨勪粙缁嶃€侟/p>
鍏嶇柅鑽у厜 (IF)
鍦ㄨ崸鍏夋樉寰暅鎶€鏈腑锛屽彲浠ラ€氳繃涓ょ鏂瑰紡瑙傚療鍒颁綘鐨勭洰鐨勮泲鐧斤細鍒╃敤鍐呮簮鑽у厜淇″彿锛屽嵆閫氳繃鍏嬮殕鎵嬫锛岀敤閬椾紶瀛︽柟娉曞皢鑽у厜铔嬬櫧涓庣洰鐨勮泲鐧界浉杩烇紱鎴栧埄鐢ㄨ崸鍏夋爣璁扮殑鎶椾綋鐗瑰紓鎬х粨鍚堢洰鐨勮泲鐧姐€傛湁浜涚敓鐗╁闂閲囩敤绗簩绉嶆柟娉曚細鏇存湁鐢ㄦ垨鏇存湁蹇呰銆傛瘮濡傦紝缁勭粐瀛︽牱鍝佹棤娉曚娇鐢ㄨ崸鍏夎泲鐧斤紝鍥犱负閫氬父鏉ヨ锛屾爣鏈兘鏄粠鏃犳硶淇濆瓨鑽у厜铔嬬櫧鐨勭敓鐗╀綋涓幏鍙栥€傛澶栵紝褰撴湁涓€涓湁鍔熻兘鐨勬姉浣撳彲鐢ㄦ椂锛屽厤鐤崸鍏夋硶浼氭瘮鑽у厜铔嬬櫧鎶€鏈揩寰堝锛屽洜涓哄悗鑰呭繀椤诲厛鍏嬮殕鐩殑鍩哄洜鍐嶅皢DNA杞煋鍒伴€傚綋鐨勭粏鑳炰腑銆傝崸鍏夎泲鐧界殑鍙︿竴椤瑰姡鍔垮湪浜庡叾鏈韩灞炰簬铔嬬櫧璐ㄣ€傚洜姝わ紝缁嗚優鍐呯殑杩欎簺鑽у厜铔嬬櫧鍏锋湁鐗瑰畾鐨勮泲鐧借川鐗规€э紝鍏朵細瀵艰嚧闄勭潃鐨勭洰鐨勮泲鐧借川鍙戠敓鍔熻兘绱婁贡鎴栧嚭鐜拌閲婄殑鎯呭喌銆傜劧鑰岋紝鑽у厜铔嬬櫧鎶€鏈粛鐒舵槸瑙傚療娲荤粏鑳炵殑棣栭€夋柟娉曘€侟/p>
鍏嶇柅鑽у厜娉曞埄鐢ㄤ簡鎶椾綋鍙互鍜岀浉搴旀姉鍘熺壒寮傛€х粨鍚堢殑杩欎釜鐗规€э紝瀵规瀹冭繕鏈変袱绉嶄笉鍚岀殑琛ㄧ幇褰㈠紡銆傛渶绠€鍗曠殑鏂瑰紡鏄娇鐢ㄥ彲涓庣洰鐨勮泲鐧界浉缁撳悎鐨勮崸鍏夋爣璁版姉浣撱€傝繖绉嶆柟娉曡绉颁负鈥滅洿鎺ュ厤鐤崸鍏夋硶鈥濄€侟/p>
鍦ㄥ緢澶氭儏鍐典笅锛屾垜浠彲浠ュ埄鐢ㄤ袱绉嶄笉鍚岀壒鎬х殑鎶椾綋銆傜涓€绉嶆姉浣撳彲浠ョ粨鍚堢洰鐨勮泲鐧斤紝浣嗗叾鏈韩骞舵湭杩涜鑽у厜鏍囪(涓€鎶?銆傜浜岀鎶椾綋鏈韩灏辨惡甯﹁崸鍏夋煋鏂?浜屾姉)锛屽苟涓斿彲浠ョ壒寮傛€х粨鍚堜竴鎶椼€傝繖绉嶆柟娉曡绉颁负鈥滈棿鎺ュ厤鐤崸鍏夋硶鈥濄€傝繖绉嶆柟娉曞瓨鍦ㄨ澶氫紭鍔裤€備竴鏂归潰锛屽畠浼氫骇鐢熸斁澶ф晥搴旓紝鍥犱负涓嶅彧涓€涓簩鎶楀彲浠ヤ笌涓€鎶楃浉缁撳悎銆傚彟涓€鏂归潰锛屾病鏈夊繀瑕佸缁堢敤鑽у厜鏌撴枡鏍囪鐩殑铔嬬櫧鐨勬瘡涓姉浣擄紝浣嗗彲浠ヤ娇鐢ㄥ競鍞崸鍏夋爣璁扮殑浜屾姉銆傚厤鐤崸鍏変腑骞挎硾浣跨敤鐨勮崸鍏夋煋鏂欏寘鎷 FITC銆乀RITC 鎴栦竴浜汚lexa Fluor?鏌撴枡锛屼笅鏂囧潎鏈夋彁鍙娿€侟/p>
FITC 鍜 TRITC
寮傜~姘伴吀鑽у厜绱?FITC) 鏄竴绉嶆湁鏈鸿崸鍏夋煋鏂欙紝鐩墠锛岃繖绉嶈崸鍏夋煋鏂欎粛鐢ㄤ簬鍏嶇柅鑽у厜鍜屾祦寮忕粏鑳炴湳涓€傚湪 495/517 nm 澶勶紝璇ユ煋鏂欎細浜х敓婵€鍙?鍙戝皠宄板€硷紝骞跺彲鍊熷姪寮傜~姘伴吀鐩愬弽搴斿熀鍥笌涓嶅悓鎶椾綋缁撳悎锛岃鍩哄洟鍙互鍜岃泲鐧借川涓婄殑姘ㄥ熀銆佸矾鍩恒€佸挭鍞戙€侀叒姘ㄩ叞銆佺景鍩虹瓑鍩哄洟鐩哥粨鍚堛€傝€屽畠鐨勫熀鏈垚鍒嗏€斺€ 鑽у厜绱狅紝鍏舵懇灏旇川閲忎负 332 g/mol锛屽父琚敤浣滆崸鍏夌ず韪墏銆侳ITC(389 g/mol) 鏄敤浜庤崸鍏夋樉寰暅鎶€鏈殑棣栨壒鏌撴枡锛屼笖鍏惰褰撴垚 Alexa Fluor?488 绛夊悗缁崸鍏夋煋鏂欑殑鍙戠銆傝鏌撴枡鐨勮崸鍏夋椿鎬у彇鍐充簬瀹冪殑澶у叡杞姵棣欑數瀛愮郴缁燂紝鑰岃绯荤粺鍙楄摑鑹插厜璋变腑鐨勫厜鎵€婵€鍙戙€侟/p>
缁忓父涓 FITC 鍚屾椂浣跨敤鐨勫彟涓€绉嶆煋鏂欐槸涓庡叾鐩镐技鐨凾RITC [鍥涚敳鍩虹綏涓规槑-5(6)-寮傜~姘伴吀]銆備笌 FITC 鐩稿弽锛孴RITC 骞堕潪鑽у厜绱狅紝鑰屾槸缃椾腹鏄庡鏃忕殑琛嶇敓鐗┿€傜綏涓规槑涔熷叿鏈変竴涓ぇ鐨勫叡杞姵棣欑數瀛愮郴缁燂紝姝f槸璇ョ郴缁熷紩鍙戜簡瀹冧滑鐨勮崸鍏夎涓恒€傝繕鏈変竴鐐逛笌FITC 鐩稿弽锛孴RITC (479 g/mol) 鐢辨渶澶ф尝闀夸负 550nm鐨勭豢鑹插厜璋变腑鐨勫厜鎵€婵€鍙戯紝瀹冪殑鏈€澶у彂灏勬尝闀夸负 573 nm銆備笌铔嬬櫧璐?渚嬪锛屾姉浣?缁撳悎涔熷熀浜庡紓纭鞍閰哥洂鍙嶅簲鍩哄洟銆侟/p>
铏界劧 FITC 鍜 TRITC 浠嶅湪浣跨敤锛屼絾鐢变簬瀹冧滑灞炰簬鍙戝厜鐩稿杈冨急鐨勮崸鍏夋煋鏂欎笖瀹冧滑鐨勪紭鍔夸粎浠呮槸缁忔祹瀹炴儬锛屽洜姝わ紝鍦ㄦ渶鏂扮殑鏄惧井闀滄妧鏈腑骞朵笉鎺ㄨ崘銆侟/p>
闈掕壊绱燑/strong>
杩欑被鑽у厜鏌撴枡鐩稿杈冨皯锛屼粠闈掕壊绱犺鐢熻€屾潵锛屼篃鏄叾鍚嶇О鐨勭敱鏉ワ細Cy2銆丆y3銆丆y5 鍜孋y7銆備笂杩版墍鏈夐潚鑹茬礌鍧囧彲浠ラ€氳繃鍏跺弽搴斿熀鍥笌鏍搁吀鎴栬泲鐧界浉杩炪€備緥濡傦紝閲囩敤浜嗚泲鐧芥爣璁扮殑椹潵閰颁簹鑳哄熀鍥€€傛湁瓒g殑鏄紝瀵逛簬鑽у厜锛孋y5 瀵瑰叾鍛ㄨ竟鐢靛瓙鐜闈炲父鏁忔劅锛岃鐗瑰緛鍙敤浜庨叾娴嬪畾銆傞檮鐫€铔嬬櫧璐ㄧ殑鏋勮薄鏀瑰彉浼氬鑷磋崸鍏夊彂灏勪骇鐢熼槼鎬ф垨闃存€у彉鍖栥€傛澶栵紝Cy3 鍜 Cy5 杩樺彲鐢ㄤ簬 FRET 璇曢獙銆傞潚鑹茬礌鏌撴枡鏄竴绉嶇浉瀵硅緝鑰佺殑鑽у厜鏌撴枡锛屼絾鍗存槸鍏朵粬鑽у厜鏌撴枡鍦ㄤ寒搴︺€佽€愬厜鎬с€侀噺瀛愪骇鐜囩瓑鏂归潰寰椾互鏀瑰杽鐨勫熀纭€銆侟/p>
Alexa Fluor?鏌撴枡
Alexa Fluor?鏌撴枡鏄甫璐熺數鑽蜂笖浜叉按鐨勮崸鍏夋煋鏂欑郴鍒楋紝璇ョ郴鍒楁煋鏂欏泭鎷寖鍥磋緝骞匡紝涓旂粡甯哥敤浜庤崸鍏夋樉寰暅鎶€鏈箣涓€傝繖浜涙煋鏂欑殑鍚嶇О鏄敱鍏跺彂鏄庤€匯ichard Paul Haugland 浠ヤ粬鍎垮瓙 Alex Haugland 鐨勫悕瀛楀懡鍚嶇殑銆傝浜у搧鏍囪瘑鏄 Molecular Probes(缇庡浗鐢熷懡绉戝鎶€鏈叕鍙 Life Technologies鏃椾笅瀛愬叕鍙革紝娉細2014骞?鏈圠ife Technologies琚玊hermo Fisher鏀惰喘)鐨勫晢鏍囥€傛澶栵紝杩欎簺浜у搧鏍囪瘑涓篃娑电洊浜嗙浉搴旂殑婵€鍏夋縺鍙戞尝闀裤€備緥濡傦紝搴旂敤鑼冨洿寰堝箍涓旀渶澶ф縺鍙戞尝闀夸负 493 nm鐨凙lexa Fluor?488锛屽彲鐢辨爣鍑嗙殑488 nm婵€鍏夋縺鍙戙€侫lexa Fluor?488鐨勬渶澶у彂灏勬尝闀夸负 519 nm锛屾鏄洜涓哄叿澶囦笂杩扮壒鎬э紝浣垮緱 Alexa Fluor?488涓 FITC 鐨勫睘鎬х浉浼笺€傚敖绠 Alexa Fluor?488鏄竴绉嶈崸鍏夌礌琛嶇敓鐗╋紝浣嗕笌 FITC 鐩稿弽锛屽畠鎷ユ湁鏇翠匠鐨勭ǔ瀹氭€у拰鑽у厜浜害锛屼笖 pH 鏁忔劅搴︿篃鏇翠綆銆傛墍鏈 Alexa Fluor? 鏌撴枡(姣斿锛孉lexa Fluor?546銆丄lexa Fluor?633)閮芥槸涓嶅悓鍩虹鑽у厜鐗╄川鐨勭:鍖栧舰寮忥紝渚嬪锛岃崸鍏夌礌銆侀璞嗙礌銆侀潚鑹茬礌鎴栫綏涓规槑锛屽畠浠殑鎽╁皵璐ㄩ噺鍦?10 鑷 1400 g/mol 鑼冨洿涔嬪唴銆侟/p>
DNA 鏌撹壊
鍦ㄨ崸鍏夋樉寰暅鎶€鏈腑锛屼笉鍙爺绌惰泲鐧界粨鏋勶紝鏍搁吀鍚屾牱鍏锋湁閲嶈鐨勭爺绌舵剰涔夈€傛湁鏃跺€欙紝蹇呴』閫氳繃妫€娴嬬粏鑳炴牳鏉ョ‘瀹氱粏鑳炵殑绮剧‘浣嶇疆鍙婂叾鏁伴噺銆傛渶甯哥敤鐨勪竴绉岲NA 鏌撹壊鍓傚綋灞 DAPI (4',6-浜岃剴鍩?2-鑻熀鍚插摎) 锛屽叾鍙笌DNA 鍙岃灪鏃嬬殑 A-T 瀵岄泦鍖哄煙鐩哥粨鍚堛€傚鏋 DAPI 闄勭潃鍒 DNA 涓婏紝鍏惰崸鍏夊己搴﹀皢姣旀父绂荤姸鎬佽楂樸€傝鏌撹壊鍓傚彈鏈€澶ф尝闀夸负358 nm鐨勭传澶栧厜婵€鍙戯紝鍏跺彂灏勫厜璋遍潪甯稿锛屽湪461 nm 澶勮揪鍒板嘲鍊笺€傛澶栵紝杩樺彲瀵瑰急鑽у厜杩涜 RNA 缁撳悎妫€娴嬨€傚湪杩欑鎯呭喌涓嬶紝鍙戝皠娉㈤暱灏嗚浆绉昏嚦 500 nm銆傛湁瓒g殑鏄紝DAPI 鑳藉绌块€忔暣涓粏鑳炶啘銆傚洜姝わ紝瀹冨彲浠ョ敤浜庡浐瀹氬拰娲荤粏鑳炰箣涓€侟/p>
绗簩绉嶈骞挎硾浣跨敤鐨凞NA 鏌撹壊鍓傚氨鏄 Hoechst 鏌撴枡绯诲垪锛岃繖浜涙煋鏂欏師鍏堥兘鏄敱 Hoechst AG 杩欏鍖栧鍏徃鐢熶骇鐨勩€侶oechst 33258銆丠oechst 33342浠ュ強Hoechst 34580 鍧囦负鍙岃嫰閰颁簹鑳猴紝鍙祵鍏 A-T 瀵岄泦鍖哄煙锛屽洜姝わ紝璇ョ郴鍒楁煋鏂欏緢灏戠敤鍒般€備笌 DAPI 鐩镐技锛岃繖涓夌鏌撹壊鍓傞兘鍙彈鏈€澶у彂灏勬尝闀夸负455 nm鐨勭传澶栧厜婵€鍙戯紝鑰屾湭琚粨鍚堢殑鏌撹壊鍓傦紝鍏舵渶澶у彂灏勬尝闀垮湪 510鈥?40 nm 涔嬮棿銆侶oechst 鏌撹壊鍓傝繕鍏锋湁缁嗚優娓楅€忔€э紝鍥犳鍙敤浜庢椿缁嗚優鎴栧凡鍥哄畾鐨勭粏鑳炰腑銆傝鏌撹壊鍓備笌DAPI 鐨勪笉鍚屼箣澶勫湪浜庯紝瀹冧滑鐨勬瘨鎬ц緝浣庛€侟/p>
纰樺寲涓欏暥(Propidium-Iodide锛孭I)鏄竴绉嶄笉鑳介€忚繃缁嗚優鑶滅殑 DNA 鏌撹壊鍓傘€傜敱浜庡叿澶囦笂杩扮壒鎬э紝璇ユ煋鑹插墏鏃犳硶杩涘叆瀹屾暣鐨勭粏鑳炰腑锛屽洜姝わ紝璇ユ煋鑹插墏甯哥敤浜庡尯鍒嗙粏鑳炵兢涓殑娲荤粏鑳炲拰姝荤粏鑳炪€傛澶栵紝纰樺寲涓欏暥杩樻槸涓€绉嶅祵鍏ュ墏锛屼絾瀵逛簬涓嶅悓鐨勭⒈鍩哄苟涓嶅瓨鍦ㄧ粨鍚堢殑宸紓鎬с€傝鏌撹壊鍓備笌鏍搁吀缁撳悎鍚庯紝鏈€澶ф縺鍙戞尝闀夸负538 nm锛屾渶澶у彂灏勬尝闀夸负 617 nm銆傛湭缁撳悎 PI 鐨勬渶澶ф縺鍙戝拰鍙戝皠娉㈤暱鍜屽厜寮轰細鏇翠綆涓€浜涖€侾I 杩樺彲缁撳悎 RNA锛屽悓鏃舵棤闇€鏀瑰彉鑷韩鐨勮崸鍏夌壒鎬с€傛湁鏃跺€欎负浜嗗尯鍒咲NA 鍜 RNA锛屾湁蹇呰浣跨敤閫傚綋鐨勬牳閰搁叾銆侟/p>
涓嶉渶瑕佸墠鏈熷鐞嗭紝鍚栧暥姗欏氨鍙互閴村埆DNA 涓 RNA 銆傚悥鍟舵涓 DNA 缁撳悎鍚庯紝鏈€澶ф縺鍙?鍙戝皠娉㈤暱涓 502 nm/525 nm锛岃€屼笌 RNA 缁撳悎鍚庯紝鏈€澶ф縺鍙?鍙戝皠娉㈤暱涓?60 nm/650 nm銆傛澶栵紝瀹冭繕鑳藉杩涘叆婧堕叾浣撶瓑閰告€у尯瀹ゃ€傚湪杩欓噷锛岄槼绂诲瓙鏌撴枡琚川瀛愬寲銆傚湪杩欑閰告€х幆澧冧笅锛屽悥鍟舵鐢辫摑鑹插厜璋变腑鐨勫厜婵€鍙戯紝浣嗗彂灏勬尝闀垮湪姗欒壊鍖哄煙杈惧埌鏈€澶с€傜敱浜庡噵浜$粏鑳炲叿鏈夊ぇ閲忚鍚炲櫖鐨勯吀鎬у尯瀹わ紝鍥犳锛屽悥鍟舵甯哥敤浣滄绫荤粏鑳炵殑鏍囪鐗┿€侟/p>
鍖哄鍜岀粏鑳炲櫒鐗瑰紓鎬ф煋鏂橖/strong>
鍦ㄨ崸鍏夋樉寰暅鎶€鏈腑锛屽線寰€瑕佸婧堕叾浣撱€佹牳鍐呬綋绛夌粏鑳炲尯瀹や互鍙婄嚎绮掍綋绛夌粏鑳炲櫒杩涜鏌撹壊銆備负姝わ紝璇ラ儴鍒嗕粙缁嶄簡涓€绯诲垪鍙緵閫夋嫨浣跨敤鐨勭壒寮傛€ф煋鏂欍€侟/p>
瑙傚療绾跨矑浣撴渶甯哥敤鐨勬柟娉曞氨鏄埄鐢 MitoTracker?锛屽畠鏄竴绉嶅彲閫忚繃缁嗚優鐨勬煋鏂欙紝鍖呭惈杞诲害宸熀鍖栫殑姘敳鍩烘椿鎬ч儴鍒嗐€傛鍥犲姝わ紝瀹冨彲涓庡崐鑳辨皑閰告畫鍩虹殑娓哥纭唶鍩哄弽搴旓紝浠庤€屼笌鍩鸿川铔嬬櫧瀹炵幇鍏变环缁撳悎銆侻itoTracker? 鏈変笉鍚岀殑棰滆壊鍜屼慨楗扮被鍨?鍙傝琛 1)锛屾澶栵紝瀹冭繕鏄 Molecular Probes 鐨勫晢鏍囥€備笌缃椾腹鏄 123 (Rh123) 鎴 tetramethylrosamine绛夊父瑙勭嚎绮掍綋鐗瑰紓鎬ф煋鑹插墏涓嶅悓锛屽湪鐢ㄥ浐瀹氬墏鐮村潖鑶滅數浣嶅悗锛孧itoTracker?涓嶄細琚礂鎺夈€侟/p>
渚濇嵁绾跨矑浣撴煋鑹插墏锛岃繕鏈変簺鏌撴枡鍙互鏍囪婧堕叾浣撶瓑閰告€у尯瀹わ紝杩欑被鏌撴枡琚О涓篖ysoTracker銆傚畠浠敱杩炴帴涓€涓崸鍏夊熀鍥㈢殑寮辩⒈鍩哄洟缁勬垚锛屽叿鏈夎啘绌块€忔€с€傛渶鏈夊彲鑳界殑鎯呭喌鏄紝杩欎簺纰卞熀鍥犺川瀛愬寲浣滅敤鐨勫奖鍝嶈€屽閰告€у尯瀹ゅ叿鏈変翰鍜屾€с€侺ysoTracker鍏锋湁澶氱涓嶅悓鐨勯鑹插彲渚涢€夋嫨(鍙傝琛 1)銆侟/p>
涓庢憾閰朵綋鐩镐技鐨勫尯瀹ゆ槸閰块厭閰垫瘝绛夌湡鑿屼腑鐨勬恫娉★紝杩欑鑶滃瘑闂┖闂翠篃鏄竴绉嶉吀鎬х幆澧冦€傚鏋滆鍦ㄨ崸鍏夋樉寰暅涓嬭瀵熶笂杩板尯瀹わ紝鍒欒浣跨敤FM 4-64?鎴朏M 5-95?绛夎嫰涔欑儻鍩烘煋鏂欍€侟/p>
瀵逛簬铔嬬櫧璐ㄥ垎娉屽疄楠岋紝鍐呰川缃 (ER) 鍏锋湁閲嶈鐨勭爺绌舵剰涔夈€傚涓婅堪鍖哄杩涜鏌撹壊鐨勪竴绉嶅吀鍨嬫煋鏂欎负DiOC6(3)銆傝鏌撴枡铏界劧鍋忓ソ ER锛屼絾浠嶄細缁撳悎绾跨矑浣撶瓑鍏朵粬缁嗚優鍣ㄨ啘銆傚ER 杩涜鐗瑰紓鎬ф煋鑹茬殑鍙︿竴绉嶆柟娉曟槸锛屼娇鐢 ER-Tracker Green 鍜 Red 绛 ER-Tracker锛岃€ ER-Tracker Green 鍜 Red 鏄熀浜 BODIPY 鐨勪袱绉嶆煋鏂欙紝鍏朵笌鏍煎垪鏈劜(涓€绉嶇:閰板熀鑴查叾)杩炴帴锛屽苟鍙笌浠呭瓨浜庡唴璐ㄧ綉鑶滀笂鐨凙TP鏁忔劅鎬ч捑閫氶亾缁撳悎銆侭ODIPY(纭?浜屽悺鍜簹鐢插熀锛宐oron-dipyrromethene)鏄竴绉嶅嚑涔庝笉婧朵簬姘寸殑銆佸pH 鐩稿涓嶆晱鎰熺殑鏌撴枡鍩哄洟锛岃鏌撴枡瀵硅泲鐧借川鏍囪娌″お澶х敤澶勶紝浣嗗嵈鏄剛璐ㄥ拰鑶滄爣璁扮殑鑹ソ宸ュ叿銆侟/p>
瀵逛簬涓嶦R鐩搁偦鐨勯珮灏斿熀浣擄紝鍙互鐢 NBD C6-ceramide 鍜孊ODIPYFL C5-ceramide 绛夎崸鍏夌缁忛叞鑳虹被浼肩墿瀵瑰叾杩涜鏍囪銆 涓婅堪绁炵粡閰拌兒涓洪灅鑴傜被锛屽叾鍦ㄩ珮灏斿熀浣撲腑楂樺害瀵岄泦銆侟/p>
鍊熷姪鍩轰簬鑴傝川鐨勬煋鏂欙紝鍙互瀵硅剛绛忕瓑鐗瑰紓鑶滃尯鍩熻繘琛屾煋鑹层€備娇鐢 NBD-6Cholestrol鎴朜BP-12 Cholesterol 鍙互瑙傚療鑳嗗浐閱囧瘜闆嗗尯鍩?Avanti Polar Lipids)銆侟/p>
闄や簡浣跨敤鐗瑰紓鎬ч潪铔嬬櫧鑽у厜鏌撴枡瀵圭粏鑳炲尯瀹よ繘琛屾爣璁颁箣澶栵紝杩樺彲浠ュ€熷姪瀵圭粏鑳炰腑涓嶅悓浣嶇疆鏈夊亸濂界殑铔嬬櫧璐ㄥ鐩爣鍖哄煙杩涜鏌撹壊銆傝繖浜涜泲鐧借川鍙互鍜岃崸鍏夋煋鏂欑浉杩烇紝骞跺彲閫氳繃鑽у厜鏄惧井闀滆繘琛岃瀵熴€傝繍鐢ㄨ繖绉嶆柟娉曠殑涓€涓疄渚嬫槸锛氶害鑳氬嚌闆嗙礌(WGA) 鍙互涓庣粏鑳炶川鑶滀腑鐨勫斁娑查吀鍜孨-涔欓叞钁¤悇绯栬兒鍩虹壒寮傛€х粨鍚堬紝灏哤GA 涓庤崸鍏夋煋鏂欏伓鑱旓紝杩欐牱鎴戜滑灏卞彲浠ヨ瀵熷埌缁嗚優璐ㄨ啘浜嗐€侟/p>
绂诲瓙鎴愬儚
鍦ㄦ湁鍏崇缁忓厓鏂归潰鐨勭爺绌朵腑锛岃瀵熷熀鍥犳椿鎬ф垨缁嗚優杩愬姩绛夊浜庝簡瑙g粏鑳炵殑绂诲瓙娴撳害鍏锋湁閲嶈鎰忎箟銆傞挔銆侀挋銆佹隘鎴栭晛绂诲瓙瀵瑰緢澶氫笉鍚岀殑缁嗚優娲诲姩閮藉叿鏈夎緝澶у奖鍝嶃€備竴鑸儏鍐典笅锛屽€熷姪鑽у厜鏍囪鐨勮灟鍚堝墏鍙皢绂诲瓙鍥颁綇锛岃灟鍚堝墏鍦ㄧ粨鍚堢浉搴旂瀛愬悗浼氭敼鍙樼瀛愮殑鍏夎氨鐗规€с€備緥濡傦紝鍒╃敤璇ュ師鐞嗙殑閽欐寚绀哄墏fura-2銆乮ndo-1銆乫luo-3銆乫luo-4鍜孋alcium-Green 绛夈€侟/p>
瀵逛簬閽犵瀛愮殑妫€娴嬶紝閫氬父浣跨敤 SBFI (sodium-bindingbenzofurzanisophthalate) 鎴朣odium Green銆侾BFI(potassium-binding benzofurzanisophthalate) 鍙互妫€娴嬮捑绂诲瓙銆侟/p>
鏈夎叮鐨勬槸锛岃繕瀛樺湪浠ヨ泲鐧借川涓轰富鐨勯挋鎸囩ず鍓傦紝鍏朵腑涓€绉嶆槸鍩轰簬姘存瘝鍖栧鍙戝厜铔嬬櫧鈥旀按姣嶇礌銆傛按姣嶇礌銆佸彂鍏変綋鑵旇偁绱犮€佸垎瀛愭哀鍜 Ca2+鐨勭浉浜掍綔鐢ㄤ細閲婃斁钃濆厜锛岃繖鏄湪鑽у厜铔嬬櫧璐ㄧ殑鍙戠幇杩囩▼涓潪甯歌憲鍚嶇殑鏈哄埗銆侟/p>
鑽у厜鏌撴枡鍙婂叾婵€鍙戝拰鍙戝皠娉㈤暱宄板€稽/strong>
涓婃枃鎻愬埌鐨勬墍鏈夋煋鏂欏湪涓嬭〃涓潎鏈夌粺璁°€傛澶栵紝杩樻兜鐩栦簡鍏朵粬鑽у厜鏌撴枡鍙婂叾婵€鍙戝拰鍙戝皠娉㈤暱宄板€笺€侟/p>
鑽у厜鏌撴枡绀轰緥 |
婵€鍙戝厜 |
Indo-1, Ca saturated |
331 nm |
Indo-1 Ca2+ |
346 nm |
Cascade Blue BSA pH 7.0 |
401 nm |
Cascade Blue |
398 nm |
LysoTracker Blue |
373 nm |
Alexa 405 |
401 nm |
LysoSensor Blue pH 5.0 |
374 nm |
LysoSensor Blue |
374 nm |
DyLight 405 |
399 nm |
DyLight 350 |
332 nm |
BFP (Blue Fluorescent Protein) |
380 nm |
Alexa 350 |
343 nm |
7-Amino-4-methylcoumarin pH 7.0 |
346 nm |
Amino Coumarin |
345 nm |
AMCA conjugate |
347 nm |
Coumarin |
360 nm |
7-Hydroxy-4-methylcoumarin |
360 nm |
7-Hydroxy-4-methylcoumarin pH 9.0 |
361 nm |
6,8-Difluoro-7-hydroxy-4-methylcoumarin pH 9.0 |
358 nm |
Hoechst 33342 |
352 nm |
Pacific Blue |
404 nm |
Hoechst 33258 |
352 nm |
Hoechst 33258-DNA |
352 nm |
Pacific Blue antibody conjugate pH 8.0 |
404 nm |
PO-PRO-1 |
434 nm |
PO-PRO-1-DNA |
435 nm |
POPO-1 |
433 nm |
POPO-1-DNA |
433 nm |
DAPI-DNA |
359 nm |
DAPI |
358 nm |
Marina Blue |
362 nm |
SYTOX Blue-DNA |
445 nm |
CFP (Cyan Fluorescent Protein) |
434 nm |
eCFP (Enhanced Cyan Fluorescent Protein) |
437 nm |
1-Anilinonaphthalene-8-sulfonic acid (1,8-ANS) |
375 nm |
Indo-1, Ca free |
346 nm |
1,8-ANS (1-Anilinonaphthalene-8-sulfonic acid) |
375 nm |
BO-PRO-1-DNA |
462 nm |
BOPRO-1 |
462 nm |
BOBO-1-DNA |
461 nm |
SYTO 45-DNA |
451 nm |
evoglow-Pp1 |
448 nm |
evoglow-Bs1 |
448 nm |
evoglow-Bs2 |
448 nm |
Auramine O |
431 nm |
DiO |
487 nm |
LysoSensor Green pH 5.0 |
447 nm |
Cy 2 |
489 nm |
LysoSensor Green |
447 nm |
Fura-2, high Ca |
336 nm |
Fura-2 Ca2+sup> |
336 nm |
SYTO 13-DNA |
488 nm |
YO-PRO-1-DNA |
491 nm |
YOYO-1-DNA |
491 nm |
eGFP (Enhanced Green Fluorescent Protein) |
488 nm |
LysoTracker Green |
503 nm |
GFP (S65T) |
489 nm |
BODIPY FL, MeOH |
502 nm |
Sapphire |
396 nm |
BODIPY FL conjugate |
503 nm |
MitoTracker Green |
490 nm |
MitoTracker Green FM, MeOH |
490 nm |
Fluorescein 0.1 M NaOH |
493 nm |
Calcein pH 9.0 |
494 nm |
Fluorescein pH 9.0 |
490 nm |
Calcein |
493 nm |
Fura-2, no Ca |
367 nm |
Fluo-4 |
494 nm |
FDA |
495 nm |
DTAF |
495 nm |
Fluorescein |
495 nm |
Fluorescein antibody conjugate pH 8.0 |
493 nm |
CFDA |
495 nm |
FITC |
495 nm |
Alexa Fluor 488 hydrazide-water |
493 nm |
DyLight 488 |
493 nm |
5-FAM pH 9.0 |
492 nm |
FITC antibody conjugate pH 8.0 |
495 nm |
Alexa 488 |
493 nm |
Rhodamine 110 |
497 nm |
Rhodamine 110 pH 7.0 |
497 nm |
Acridine Orange |
431 nm |
Alexa Fluor 488 antibody conjugate pH 8.0 |
499 nm |
BCECF pH 5.5 |
485 nm |
PicoGreendsDNA quantitation reagent |
502 nm |
SYBR Green I |
498 nm |
Rhodaminen Green pH 7.0 |
497 nm |
CyQUANT GR-DNA |
502 nm |
NeuroTrace 500/525, green fluorescent Nissl stain-RNA |
497 nm |
DansylCadaverine |
335 nm |
Rhodol Green antibody conjugate pH 8.0 |
499 nm |
Fluoro-Emerald |
495 nm |
Nissl |
497 nm |
Fluorescein dextran pH 8.0 |
501 nm |
Rhodamine Green |
497 nm |
5-(and-6)-Carboxy-2', 7'-dichlorofluorescein pH 9.0 |
504 nm |
DansylCadaverine, MeOH |
335 nm |
eYFP (Enhanced Yellow Fluorescent Protein) |
514 nm |
Oregon Green 488 |
498 nm |
Oregon Green 488 antibody conjugate pH 8.0 |
498 nm |
Fluo-3 |
506 nm |
BCECF pH 9.0 |
501 nm |
SBFI-Na+ |
336 nm |
Fluo-3 Ca2+ |
506 nm |
Rhodamine 123, MeOH |
507 nm |
FlAsH |
509 nm |
Calcium Green-1 Ca2+ |
506 nm |
Magnesium Green |
507 nm |
DM-NERF pH 4.0 |
493 nm |
Calcium Green |
506 nm |
Citrine |
515 nm |
LysoSensor Yellow pH 9.0 |
335 nm |
TO-PRO-1-DNA |
515 nm |
Magnesium Green Mg2+ |
507 nm |
Sodium Green Na+ |
507 nm |
TOTO-1-DNA |
514 nm |
Oregon Green 514 |
512 nm |
Oregon Green 514 antibody conjugate pH 8.0 |
513 nm |
NBD-X |
466 nm |
DM-NERF pH 7.0 |
509 nm |
NBD-X, MeOH |
467 nm |
CI-NERF pH 6.0 |
513 nm |
Alexa 430 |
431 nm |
Alexa Fluor 430 antibody conjugate pH 7.2 |
431 nm |
CI-NERF pH 2.5 |
504 nm |
Lucifer Yellow, CH |
428 nm |
LysoSensor Yellow pH 3.0 |
389 nm |
6-TET, SE pH 9.0 |
521 nm |
Eosin antibody conjugate pH 8.0 |
525 nm |
Eosin |
524 nm |
6-Carboxyrhodamine 6G pH 7.0 |
526 nm |
6-Carboxyrhodamine 6G, hydrochloride |
525 nm |
Bodipy R6G SE |
528 nm |
BODIPY R6G, MeOH |
528 nm |
6 JOE |
520 nm |
Cascade Yellow antibody conjugate pH 8.0 |
399 nm |
Cascade Yellow |
399 nm |
mBanana |
540 nm |
Alexa Fluor 532 antibody conjugate pH 7.2 |
528 nm |
Alexa 532 |
528 nm |
Erythrosin-5-isothiocyanate pH 9.0 |
533 nm |
6-HEX, SE pH 9.0 |
534 nm |
mOrange |
548 nm |
mHoneydew |
478 nm |
Cy 3 |
549 nm |
Rhodamine B |
543 nm |
DiI |
551 nm |
5-TAMRA-MeOH |
543 nm |
Alexa 555 |
553 nm |
Alexa Fluor 555 antibody conjugate pH 7.2 |
553 nm |
DyLight 549 |
555 nm |
BODIPY TMR-X, SE |
544 nm |
BODIPY TMR-X, MeOH |
544 nm |
PO-PRO-3-DNA |
539 nm |
PO-PRO-3 |
539 nm |
Rhodamine |
551 nm |
Bodipy TMR-X conjugate |
544 nm |
POPO-3 |
533 nm |
Alexa 546 |
562 nm |
BODIPY TMR-X antibody conjugate pH 7.2 |
544 nm |
Calcium Orange Ca2+ |
549 nm |
TRITC |
550 nm |
Calcium Orange |
549 nm |
Rhodaminephalloidin pH 7.0 |
558 nm |
MitoTracker Orange |
551 nm |
MitoTracker Orange, MeOH |
551 nm |
Phycoerythrin |
565 nm |
Magnesium Orange |
550 nm |
R-Phycoerythrin pH 7.5 |
565 nm |
5-TAMRA pH 7.0 |
553 nm |
5-TAMRA |
549 nm |
Rhod-2 |
552 nm |
FM 1-43 |
472 nm |
Rhod-2 Ca2+ |
553 nm |
Tetramethylrhodamine antibody conjugate pH 8.0 |
552 nm |
FM 1-43 lipid |
473 nm |
LOLO-1-DNA |
568 nm |
dTomato |
554 nm |
DsRed |
563 nm |
Dapoxyl (2-aminoethyl) sulfonamide |
372 nm |
Tetramethylrhodamine dextran pH 7.0 |
555 nm |
Fluor-Ruby |
554 nm |
Resorufin |
571 nm |
Resorufin pH 9.0 |
571 nm |
mTangerine |
568 nm |
LysoTracker Red |
578 nm |
Lissaminerhodamine |
572 nm |
Cy 3.5 |
578 nm |
Rhodamine Red-X antibody conjugate pH 8.0 |
573 nm |
Sulforhodamine 101, EtOH |
578 nm |
JC-1 pH 8.2 |
593 nm |
JC-1 |
592 nm |
mStrawberry |
575 nm |
MitoTracker Red |
578 nm |
MitoTracker Red, MeOH |
578 nm |
X-Rhod-1 Ca2+ |
580 nm |
Alexa Fluor 568 antibody conjugate pH 7.2 |
579 nm |
Alexa 568 |
576 nm |
5-ROX pH 7.0 |
578 nm |
5-ROX (5-Carboxy-X-rhodamine, triethylammonium salt) |
578 nm |
BO-PRO-3-DNA |
574 nm |
BOPRO-3 |
574 nm |
BOBO-3-DNA |
570 nm |
Ethidium Bromide |
524 nm |
ReAsH |
597 nm |
Calcium Crimson |
589 nm |
Calcium Crimson Ca2+ |
590 nm |
mRFP |
585 nm |
mCherry |
587 nm |
Texas Red-X antibody conjugate pH 7.2 |
596 nm |
HcRed |
590 nm |
DyLight 594 |
592 nm |
Ethidium homodimer-1-DNA |
528 nm |
Ethidiumhomodimer |
528 nm |
Propidium Iodide |
538 nm |
SYPRO Ruby |
467 nm |
Propidium Iodide-DNA |
538 nm |
Alexa 594 |
590 nm |
BODIPY TR-X, SE |
588 nm |
BODIPY TR-X, MeOH |
588 nm |
BODIPY TR-X phallacidin pH 7.0 |
590 nm |
Alexa Fluor 610 R-phycoerythrin streptavidin pH 7.2 |
567 nm |
YO-PRO-3-DNA |
613 nm |
Di-8 ANEPPS |
469 nm |
Di-8-ANEPPS-lipid |
469 nm |
YOYO-3-DNA |
612 nm |
Nile Red-lipid |
553 nm |
Nile Red |
559 nm |
DyLight 633 |
624 nm |
mPlum |
587 nm |
TO-PRO-3-DNA |
642 nm |
DDAO pH 9.0 |
648 nm |
Fura Red, high Ca |
434 nm |
Allophycocyanin pH 7.5 |
651 nm |
APC (allophycocyanin) |
650 nm |
Nile Blue, EtOH |
631 nm |
TOTO-3-DNA |
642 nm |
Cy 5 |
646 nm |
BODIPY 650/665-X, MeOH |
646 nm |
Alexa Fluor 647 R-phycoerythrin streptavidin pH 7.2 |
569 nm |
DyLight 649 |
652 nm |
Alexa Fluor 647 antibody conjugate pH 7.2 |
653 nm |
Alexa 647 |
653 nm |
Fura Red Ca2+ |
435 nm |
Atto 647 |
644 nm |
Fura Red, low Ca |
472 nm |
Carboxynaphthofluorescein pH 10.0 |
600 nm |
Alexa 660 |
664 nm |
Alexa Fluor 660 antibody conjugate pH 7.2 |
663 nm |
Cy 5.5 |
673 nm |
Alexa Fluor 680 antibody conjugate pH 7.2 |
679 nm |
Alexa 680 |
679 nm |
DyLight 680 |
678 nm |
Alexa Fluor 700 antibody conjugate pH 7.2 |
696 nm |
Alexa 700 |
696 nm |
FM 4-64, 2% CHAPS |
506 nm |
FM 4-64 |
508 nm |