daoyan：jizhifuzhujiguangjiexidianlizhipuchengxiangjishu（maldi-msi）fazhanzhijin，shoudaoguojijiguoneizhongduozhuanjiaxuezhedeguanzhuyurenke。gaijishujinenggoukeshihuachengxianmubiaohuahewuzaizuzhihexibaozhongdeerweilizimidutu，hainenggoutongshichengxianzhongduoqitahuahewudezucheng、xiangduifengdujikongjianfenbuqingkuang，shixianduishengwuyangbendewubiaoji、gaotongliangjiance。zidongjujiao3d ap-smaldizhipuchengxiangjishudehengkongchushiweifeipingmianshengwuyangbendejiancetigonglegengweigaoxiaoheqiangyoulidejishuzhicheng，kezhijieyongyuzhiwuyepian、huabandengyangbenbiaomianhuahewudekeshihuafenxi。ziranjiezhong，zhiwuzaizhenggeshengzhangzhouqineihuishoudaolaizifangfangmianmiandeweixie，rushengwuxiepohefeishengwuxiepo，qizhongshengwuxiepobaokuozhishixingdongwuyijigezhongbingyuanweishengwudeqinhai。weilemianduizhishixingdongwuzheigesudi，zhiwudefangyushouduanyekeweiduozhongduoyang，qizhongzhijiefangyucuoshijiushizhiwuzaibeiqushishichanshengyoududecishengdaixiechanwuyicishanghailaiqinhaidezhishixingdongwu，yingxiangqishengzhangfayu。

        2021nian2yue5ri，deguojisendaxuebernhard spenglerjiaoshoutuanduizaianalytical and bioanalytical chemistryshangfabiaolemingwei “3d-surface maldi mass spectrometry imaging for visualising plant defensive cardiac glycosides in asclepias curassavica ”dewenzhang，yanjiurenyuanliyong3dbiaomianmaldi-msijishu，yimalijinweiyanjiuduixiang，duimalijingandeng8zhongqiangxinganleizhiwufangyuxingwuzhijinxinglekongjianjiexi。jieguobiaoming，daliangfangyuxiangguandaixiewufenbuyujixiesunshang(monicaoshidongwugongji) malijinyepian(yepianbiaomiangaodubianhuada700μm)deshousunshangbuwei。benyanjiuchongfenzhanshilezidongjujiaoap-smaldizhipuchengxiangjishudeqiangdanengliheqianzaideshiyongxing，nenggoufenxijinshenglizhuangtaixiazhiwuyangpindesanweibiaomianhuahewufenbuxinxi。

       zaifenxishousunshangmalijinyepianzhiqian，zuozheliyong3d ap-smaldizhipuchengxiangjishufenxilewanzhengyepianshangqiangxinganleifangyuwuzhidekongjianfenbu。jieguobiaoming，zaiwanzhengyepianzhong8zhongqiangxinganleifangyuxingdaixiewujunfenbuzaizhumainei，jifenbuyuwujieruzhiguanxibaosuobaohandegaonongduruzhizhong（tu1）。

       图1（a）3D AP-SMALDI MSI检测后的马利筋叶片光学显微图像。（b）叶片表面激光烧蚀斑放大图像。（c）完整马利筋叶片3D光学显微图像，叶片表面高度变化高达350μm(图示蓝色到红色表示叶片表面高度越来越高。d RG叠加的MSI图像，马利筋苷(m/z613.2410，[M+K]+，红色)和二糖(m/z381.0793[M+H]+，绿色)。（e）congyepianyemaiquyuhuodedezhiliangfanweiweim/z420-620dedanxiangsuzhiputu。msituxiang178×178pixels，kongjianfenbianlv45μm，bilichia、c、dwei1mm、bwei150μm。

       suihouzuozhejiancelejixiesunshang1xiaoshihoudemalijinyepian，jieguoxianshi，qiangxinganleifangyuxingdaixiewujunjizhongfenbuyushousunbuwei（tu2）。youyuzaiyepianshousun10minhouqishangkouchujikejiancedaogaonongduqiangxingan（tu3a-c），guyongqiangxinganhechengzengduolaijieshizheiyixianxiangmaosibingbuheli。yinci，benyanjiuyuyiwangyanjiuyizhirenweiruzhishiyizhongkefenpeidefangyuwuzhi，jiyepianshousunshanghouruzhizaidiyishijianyunshuzhishoushangbuwei，shideyoudufangyuhuahewuzaisunshangbuweixunsufuji。

       图2（a）3D AP-SMALDI MSI检测后的马利筋叶片光学显微图像。（b）马利筋叶片3D光学显微图像，损伤部位的高度差最高可达400μm。（c）RGB叠加MSI图像，乌拉卡定(m/z569.2132，[M+K]+，红色)、二糖(m/z365.1056，[M+Na]+，绿色)和三羟基黄酮丙二酰糖苷(m/z557.0692，[M+K]+，蓝色)。（d） wulakading(m/z569.2132，[m+k]+)。（e）ertang(m/z365.1056，[m+na]+)。（f）sanqiangjihuangtongbingerxiantanggandanlizituxiang(m/z557.0692，[m+k]+)。msituxiang143×171 pixels，kongjianfenbianlv35μm，suoyoubilichijunwei1mm。

       jinyibuzuozhekaochalefangyudaixiewubutongshijiandiandejileisulv，zuozhefenbiejianceleshangkoujiangeshijianwei4h50min（jiyepianshousunshang5hhe10minhoujinxingjiance）he22h（jiyepianshousunshang24hhe2hhoujinxingjiance）dejixiesunshangyepian。jieguoxianshi，jiangeshijianjiaoduandelianggeshangkouchuqiangxinganleiwuzhidejileiliangxiangchabuda，erjiangeshijianjiaozhangdelianggeshangkouchuzechengxianchumingxiandechayi。yepianshousun24xiaoshihou，shousunbuweiqiangxinganleiwuzhimingxianjianshao，biaominggaileiwuzhisuizheshijiandeyanzhangfashenglezhuanyihuojiangjie（tu3）。

       图3（a,d）AP-SMALDI MSI检测后叶片3D光学显微图像，对受伤区域进行标记，并标出相关的时间间隔。（b,e）3D显微图像，叶片表面高度差分别达500μm和600μm。（c,f）RGB叠加MSI图像calotoxin(m/z587.2250，[M+K]+，红色)，二羟基黄酮(m/z255.0652，[M+H]+，绿色)和m/z255.2110(蓝色) msituxiangc111×111 pixels，kongjianfenbianlv45μm，f112×103 pixels，kongjianfenbianlv35μm，bilichiwei1mm。

       zhiwuyoufangyushouduan，dongwuyiyoubushicelve。xuduozhishixingkunchongtongguomaiqiedefangshilaizuzhizhiwufangyu。liru，bushishikunchongxianqieduanyemaishangduan，shifangyuwuzhibunengzaizhuanyibingjileizaixiayoubuwei。yinci，zuozhecaiyong3d ap-smaldi msijishuduimaiqieyepiandejinxinduan（qiekoushangyou）jiyuanxinduan（qiekouxiayou）shangkoujinxingjiance，faxianyuanxinduanbijinxinduanfangyuxingdaixiewujileimingxianjianshao，shuomingmaiqieyingxiangxianguscharinzheileidefangyuxingwuzhidezhuanyishiqibunengdaodaxiayouyepianfahuiqifangyuzuoyong，jieshileweishenmedaduoshuhanyouruzhidezhiwu(rumalijin)dezhuanmenbushizhezaiqushiyepianshicaiyongmaiqiezuoweiyizhongyufangzhongdudecelve。

       图4（a）AP-SMALDI MSI检测前叶片表面的光学显微图像，叶片表面渗出乳汁的区域被红线标记出来。（b）叶片3D光学显微图像，高度差高达700μm。（c, d）RGB叠加MSI图像Uscharin(m/z626.2189，[M+K]+，红色)，（d）脱镁叶绿素a(m/z909.5291，[M+K]+，红色)。m/z771.4869(蓝色)和丙二酰染料木甙(m/z557.0692，[M+K]+，绿色)shimubiaohuahewuc、ddebeijingxinhao。msituxiang157×171pixels，kongjianfenbianlv40μm，bilichiwei1mm。

       jielun：benwencaiyongzidongjujiao3d ap-smaldi msijishuduimalijinyepianzhongfangyuxingdaixiewujinxinglejingquedingwei，biaomingruzhishizhiwudiyuzuzhisunshangheqianzaizhishixingkunchonggongjidehuaxuefangyujizhideyibufen，zhiwuyepianshousunhou，ruzhiliuxiangshangkoudesuduzengjia，shidefangyuwuzhizaiyepianshousunbuweijilei, bingjinyibufenxilezheizhongjileisulvzaishijianhekongjianshangdebianhua。ciwai，yanjiurenyuanmofangkunchongdebushicelve（maiqie）laidiyuzheizhongfangyujizhi，guanchadaozaiqiegeshangkoudeyuanduanyoujiaoshaodeyoududaixiewujilei。tongshi，daliangweishibiedelizixinhaoxianshichuyuqiangxinganleiwuzhijuyouxiangtongdekongjianfenbu，biaomingmalijinzhongcunzaizheduozhongduoyangdefangyuxingwuzhi。yinci，zaimaldi zhipuchengxiangdebeijingxia，yihplc-mszuoweibuchongfangfadefeibaxiangdaixiezuxueshifaxianheyanjiuzhiwuyucaoshixingkunchonghebingyuanjunxianghuzuoyongguanxidejuejiafangfa，yucitongshi，zuzhiyuanweimaldi ms/msjiancetongyangyouzhuyuduiweizhidaixiewudejianding。