MTT/鍣诲攽钃滭/span>

绠€瑕佷粙缁嶏細

MTT鍟嗗搧鍚嶅櫥鍞戣摑锛屾槸涓€绉嶉粍棰滆壊鐨勬煋鏂欍€侻TT涓昏鐢ㄩ€旓細

1锛庤嵂鐗╋紙鎴栧叾浠栧鐞嗘柟寮忥級瀵逛綋澶栧煿鍏荤殑缁嗚優娲绘€х殑娴嬪畾锛汓/span>

2锛庣粏鑳炲娈栧強缁嗚優娲绘€ф祴瀹氥€侟/span>

MTT骞挎硾鐢ㄤ簬涓€浜涚敓鐗╂椿鎬у洜瀛愮殑娲绘€ф娴嬨€佸ぇ瑙勬ā鐨勮嵂鐗╃瓫閫夈€佺粏鑳炴椿鎬ц瘯楠屼互鍙婅偪鐦ゆ晱鎰熸€ф祴瀹氱瓑銆傚畠鐨勭壒鐐规槸鐏垫晱搴﹂珮銆佺粡娴庡疄鎯犮€侟/span>

璐у彿锛欳T-MTT-1

鍖呰閲忥細1.0鍏婱TT绮夋湯

鍌ㄥ瓨锛?4掳C锛岄伩鍏堻/span>

MTT鍏ㄧО涓?-锛?,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide锛屾眽璇寲瀛﹀悕涓 3-锛?锛?-浜岀敳鍩哄櫥鍞?2锛?2锛?-浜岃嫰鍩哄洓姘攽婧寸洂锛屽晢鍝佸悕锛氬櫥鍞戣摑锛屾槸涓€绉嶉粍棰滆壊鐨勬煋鏂欍€傚湪姘翠腑鐨勬憾瑙ｅ害涓?0 mM銆侟/span>

鎶€鏈祫鏂欙細

CAS No锛?98-93-1

鍒嗗瓙閲忥細414.32

鍒嗗瓙寮忥細C18H16N5SBr

绾害锛?gt;98%

MTT涓昏鏈変袱涓敤閫旓細

1锛庤嵂鐗╋紙鎴栧叾浠栧鐞嗘柟寮忥級瀵逛綋澶栧煿鍏荤殑缁嗚優娲绘€х殑娴嬪畾锛汓/span>

2锛庣粏鑳炲娈栧強缁嗚優娲绘€ф祴瀹氥€侟/span>

MTT瀹為獙鐨勫師鐞嗭細

缁嗚優绾跨矑浣撲腑鐨勭惀鐝€閰歌劚姘㈤叾鑳藉皢MTT杩樺師涓烘按涓嶆憾鎬х殑钃濈传鑹茬粨鏅剁敳鑷紙Formazan锛夛紝骞舵矇绉湪缁嗚優涓紝鑰屾缁嗚優涓嶈兘杩涜杩欎釜鍙嶅簲銆備簩鐢插熀浜氱牅锛圖MSO锛夎兘婧惰В缁嗚優涓殑鐢茶嚔锛岀敤閰舵爣浠湪570 nm娉㈤暱澶?490 nm -570 nm涔嬮棿閮藉彲浠ワ級娴嬪畾鍏跺厜鍚告敹鍊硷紝鍦ㄤ竴瀹氱粏鑳炴暟鑼冨洿鍐咃紝MTT缁撴櫠褰㈡垚鐨勯噺涓庣粏鑳炴暟鎴愭姣斻€傛牴鎹祴寰楃殑鍚稿厜搴﹀€硷紙OD鍊硷級鏉ュ垽鏂椿缁嗚優鏁伴噺锛孫D鍊艰秺澶э紝缁嗚優娲绘€ц秺寮猴紙濡傛灉鏄祴鑽墿姣掓€э紝鍒欒〃绀鸿嵂鐗╂瘨鎬ц秺灏忥級銆傝鏂规硶宸插箍娉涚敤浜庝竴浜涚敓鐗╂椿鎬у洜瀛愮殑娲绘€ф娴嬨€佸ぇ瑙勬ā鐨勮嵂鐗╃瓫閫夈€佺粏鑳炴椿鎬ц瘯楠屼互鍙婅偪鐦ゆ晱鎰熸€ф祴瀹氱瓑銆傚畠鐨勭壒鐐规槸鐏垫晱搴﹂珮銆佺粡娴庡疄鎯犮€侟/span>

MTT瀹為獙鏂规硶锛堜緵鍙傝€冿級

涓€銆丮TT鐨勯厤鍒迭/span>

閫氬父閰嶅埗鐨凪TT娴撳害涓? mg/ml銆傚彲浠ョО鍙朚TT 0.5鍏嬶紝婧朵簬100 ml鐨勭７閰哥洂缂撳啿娑诧紙PBS锛夋垨鏃犻厷绾㈢殑鍩瑰吇鍩轰腑锛岀敤0.22 渭m婊よ啘杩囨护浠ラ櫎鍘绘憾娑查噷鐨勭粏鑿岋紝鏀?鈩 閬垮厜淇濆瓨锛屼袱鍛ㄥ唴鐢ㄥ畬銆備篃鍙垎瑁呭悗閬垮厜淇濆瓨鍦?20鈩冮暱鏈熶繚瀛橈紝骞堕伩鍏嶅弽澶嶅喕铻嶃€傚湪閰嶅埗鍜屼繚瀛樼殑杩囩▼涓紝瀹瑰櫒**鐢ㄩ摑绠旂焊鍖呬綇銆侟/span>

浜屻€佸疄楠屾楠｜/span>

A銆佽创澹佺粏鑳濣/span>

1銆佹敹闆嗗鏁版湡缁嗚優锛岃皟鏁寸粏鑳炴偓娑叉祿搴︼紝鍚?6瀛旀澘姣忓瓟鍔犲叆100ul閾烘澘锛岃皟鑺傚緟娴嬬粏鑳炲瘑搴︿娇姣忓瓟鍚?000-10000涓粏鑳烇紝杈圭紭瀛旂敤鏃犺弻姘存垨PBS濉厖銆侟/span>

2銆佸湪5%CO2锛?7鈩冩潯浠朵笅瀛佃偛銆傚師鍒欎笂锛岀粏鑳炶创澹佸悗鍗冲彲鍔犺嵂锛屾垨涓ゅ皬鏃讹紝鎴栧崐澶╂椂闂达紝鎴栬繃澶溿€備竴鑸父鍦ㄥ墠涓€澶╀笅鍗堥摵鏉匡紝娆℃棩涓婂崍鍔犺嵂銆備竴鑸5-7涓嵂鐗╂搴︼紝姣忓瓟100ul锛屼竴鑸3涓瀛斿嵆鍙紝澶嶅瓟瓒婂缁撴灉瓒婂噯纭€傚悓鏃惰缃皟闆跺瓟锛堜粎鍚煿鍏诲熀鍜岃嵂鐗╂憾瑙ｄ粙璐級鍜屽鐓у瓟锛堜粎鍚粏鑳炲拰鑽墿婧惰В浠嬭川锛夈€侟/span>

3銆佺疆浜?%CO2锛?7鈩冩潯浠朵笅锛屽鑲叉椂闂存牴鎹嵂鐗╃殑浣滅敤鐗圭偣鏉ュ喅瀹氾紝涓€鑸?4-72灏忔椂锛屼腑閫斿彲鍦ㄥ€掔疆鏄惧井闀滀笅瑙傚療缁嗚優鐘跺喌銆侟/span>

4銆佹瘡瀛斿姞鍏?0ul MTT婧舵恫锛? mg/ml锛屽嵆0.5% MTT锛夛紝37鈩冪户缁煿鍏?-4灏忔椂銆傝嫢鑽墿鑳藉涓嶮TT鍙嶅簲锛屽彲鍏堢蹇冨悗寮冨幓鍩瑰吇娑诧紝灏忓績鐢≒BS娲?-3閬嶅悗锛屽啀鍔犲叆鍚玀TT鐨勫煿鍏绘恫銆侟/span>

5銆佺粓姝㈠煿鍏伙紝灏忓績鍚稿幓瀛斿唴鍩瑰吇娑层€侟/span>

6銆佹瘡瀛斿姞鍏?50 ul DMSO锛岀疆鎽囧簥涓婁綆閫熸尟鑽?0鍒嗛挓锛屼娇缁撴櫠鐗╁厖鍒嗘憾瑙ｃ€傚湪570nm锛堝彲鐢?30nm鏍″噯锛夋祴閲忓悇瀛旂殑鍚稿厜鍊笺€侟/span>

B銆佹偓娴粏鑳濣/span>

1銆佹敹闆嗗鏁版湡缁嗚優锛岃皟鑺傜粏鑳炴偓娑叉祿搴?E6/ml锛屼緷娆″皢鈶犵粏鑳炴偓娑?0ul锛堝嵆5E4cell/瀛旓級锛屸憽寰呮娴嬭嵂鐗?0ul锛屽叡100ul鍔犲叆鍒?6瀛旀澘锛堣竟缂樺瓟鐢ㄦ棤鑿屾按鎴朠BS濉厖锛夈€備竴鑸5-7涓嵂鐗╂搴︼紝涓€鑸3涓瀛斿嵆鍙紝澶嶅瓟瓒婂缁撴灉瓒婂噯纭€傚悓鏃惰缃皟闆跺瓟锛堜粎鍚煿鍏诲熀鍜岃嵂鐗╂憾瑙ｄ粙璐級鍜屽鐓у瓟锛堜粎鍚粏鑳炲拰鑽墿婧惰В浠嬭川锛夈€侟/span>

2銆佺疆浜?% CO2锛?7 鈩冩潯浠朵笅锛屽鑲叉椂闂存牴鎹嵂鐗╃殑浣滅敤鐗圭偣鏉ュ喅瀹氾紝涓€鑸?4-72灏忔椂锛屼腑閫斿彲鍦ㄥ€掔疆鏄惧井闀滀笅瑙傚療缁嗚優鐘跺喌銆侟/span>

3銆佹瘡瀛斿姞鍏?0 ul MTT婧舵恫锛? mg/ml锛屽嵆0.5%MTT锛夛紝37鈩冪户缁煿鍏?-4 h銆侟/span>

4銆佺蹇冿紙1000杞瑇10 min锛夛紝灏忓績鍚告帀涓婃竻锛屾瘡瀛斿姞鍏?50 渭l DMSO锛岀疆鎽囧簥涓婁綆閫熸尟鑽?0 min锛屼娇缁撴櫠鐗╁厖鍒嗘憾瑙ｃ€傚湪570 nm锛堝彲鐢?30 nm鏍″噯锛夋祴閲忓悇瀛旂殑鍚稿厜鍊笺€侟/span>

娉ㄦ剰浜嬮」

1銆丮TT娉曞彧鑳界敤鏉ユ娴嬬粏鑳炵浉瀵规暟鍜岀浉瀵规椿鍔涳紝浣嗕笉鑳芥祴瀹氱粏鑳?*鏁般€傚湪鐢ㄩ叾鏍囦华妫€娴嬬粨鏋滅殑鏃跺€欙紝涓轰繚璇佸疄楠岀粨鏋滅殑鍑嗙‘锛孧TT 鍚稿厜鍊艰鎺у埗鍦ㄤ华鍣ㄧ殑绾挎€ц鍙栬寖鍥村唴銆侟/span>

2銆丮TT**鐜扮敤鐜伴厤锛岄厤鍒舵垚5 mg/ml鏃犺弻杩囨护鍚庯紝鍦? 鈩冮伩鍏変繚瀛樹袱鍛ㄥ唴鏈夋晥锛屾垨鍦?20鈩冮暱鏈熶繚瀛橈紝閬垮厤鍙嶅鍐昏瀺锛?*灏忓墏閲忓垎瑁咃紝鐢ㄩ伩鍏夎鎴栨槸榛戠焊銆侀敗绠旂焊鍖呬綇閬垮厜锛屼互鍏嶅垎瑙ｃ€侟/span>

3銆佷繚瀛樼殑MTT婧舵恫闇€瑕佹棤鑿岋紝MTT瀵硅弻寰堟晱鎰燂紱寰€96瀛旀澘鍔犳椂涓嶉伩鍏変篃娌℃湁鍏崇郴锛屾瘯绔熸椂闂磋緝鐭紝涓嶆斁蹇冪殑璇濆彲浠ユ妸鎿嶄綔鍙颁笂鐨勭収鏄庣伅鍏虫帀銆侟/span>

4銆侀厤鍒禡TT鏃剁敤PBS婧惰В,涔熸湁浜虹敤鐢熺悊鐩愭按閰嶏紝鍙敤60 鈩冩按娴村姪婧躲€侟/span>

5銆丮TT鐨勭己鐐癸細鐢变簬MTT缁忚繕鍘熸墍浜х敓鐨勭敳鑷骇鐗╀笉婧朵簬姘达紝闇€琚憾瑙ｅ悗鎵嶈兘妫€娴嬨€傝繖涓嶄粎浣垮伐浣滈噺澧炲姞锛屼篃浼氬瀹為獙缁撴灉鐨勫噯纭€т骇鐢熷奖鍝嶏紝鑰屼笖婧惰В鐢茶嚔鐨勬湁鏈烘憾鍓傚瀹為獙鑰呬篃鏈夋崯瀹炽€侟/span>

6銆丮TT鏈夎嚧鐧屾€э紝鐢ㄧ殑鏃跺€欏皬蹇冿紝涓轰簡鎮ㄧ殑瀹夊叏鍜屽仴搴凤紝璇风┛瀹為獙鏈嶅苟鎴翠竴娆℃€ф墜濂楁搷浣滐紝**鎴撮€忔槑鐨勮杽鑶淧E鎵嬪銆侟/span>

7銆佹湰浜у搧浠呴檺浜庝笓涓氫汉鍛樼殑绉戝鐮旂┒浣跨敤銆侟/span>