CN104597113A - 一种高分辨质谱成像***图像采集半导体薄膜、制备方法及应用 - Google Patents
一种高分辨质谱成像***图像采集半导体薄膜、制备方法及应用 Download PDFInfo
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Abstract
本发明属于质谱成像领域,具体涉及一种高分辨质谱成像***图像采集半导体薄膜、制备方法及应用。本发明所述高分辨质谱成像***图像采集半导体薄膜,通过如下方法制备得到:称取半导体纳米颗粒,首先置于马弗炉中灼烧,进一步用玛瑙研钵磨细,使其分散均匀,得到半导体纳米粉末;最后将半导体纳米粉末置于压片机中压制得到半导体薄膜。本发明利用半导体纳米材料的激光诱导隧道电子俘获原理使样品分子离子化,无背景干扰,克服了常规MALDI基质的局限性,所述半导体薄膜简单易得,质谱信号稳定,表面均匀光滑,不产生背景干扰,可用于指纹分析和动植物组织切片分析,特别适合于小分子物质的准确质谱成像,便于质量控制和产业化。
Description
技术领域
本发明属于质谱成像领域,具体涉及一种高分辨质谱成像***图像采集半导体薄膜、制备方法及应用。
背景技术
质辅助激光解析离解质谱是目前质谱成像常用的一种分析技术,该技术将可吸收激光能量的有机小分子基质覆盖与组织切片表面,并将能量传递给样品分子,使之汽化并离子化,再被质量分析器检测。在该技术中,有机小分子基质与样品分子的混合模式是关键,因为它直接影响分析结果的准确度、分辨率以及实验结果的重现性和定量分析的能力。
现有的技术中,常常采用有机溶剂先溶解基质,再将基质溶液喷雾在组织切片的表面,待溶剂挥发后,样品与基质分子形成混晶。现有技术的主要缺点在于很难形成大小均匀,形貌可控的晶体,从而使得激光在不同扫描时间所获得的图谱不具有重现性,信号强度与样品量之间没有定量关系。并且,由于晶体大小和形貌的差异,造成激光轰击样品分子之后所得离子的初速度和方向不同,影响图像的分辨率和质量准确度。此外,这些有机小分子基质还通常在低质量区产生一系列背景峰,抑制低质量分子信号,并严重污染离子源。
发明内容
本发明针对现有技术的不足,目的在于提供一种高分辨质谱成像***图像采集薄膜、制备方法及应用。
一种高分辨质谱成像***图像采集半导体薄膜,其是将半导体纳米颗粒灼烧去除表面附着的有机杂质后,再经研磨处理然后置于压片机中压制成膜得到的。
按上述方案,所述半导体纳米颗粒为(Bi2O3)0.07(CoO)0.03(ZnO)0.9半导体颗粒。
按上述方案,所述灼烧的温度为350℃,灼烧时间为1小时。
上述高分辨质谱成像***图像采集半导体薄膜的制备方法,包括如下步骤:
1)将半导体纳米颗粒在350℃马弗炉中灼烧1小时;
2)将步骤1)得到的半导体纳米颗粒进一步用玛瑙研钵磨细,使其分散均匀,得到半导体纳米粉末;
3)将步骤2)得到的半导体纳米粉末放入压片机的磨具,再放入压片机,施加压力压制得到半导体薄膜;
4)将步骤3)压制得半导体薄膜取出,室温保存。按上述方案,所述压制为2000kg~4800kg压力下压制1分钟。
上述高分辨质谱成像***图像采集半导体薄膜在隐形指纹图像分析、动物组织切片图像分析、植物组织切片图像分析中的应用。
按上述方案,所述的应用为:将植物组织切片,动物组织切片或隐形指纹固定或按压在上述高分辨质谱成像***图像采集半导体薄膜上后,将半导体薄膜固定在样品靶上,直接放入质谱仪进行分析。
按上述方案,所述在隐形指纹图像分析中的应用为:将指纹直接按压于半导体薄膜表面后,固定半导体薄膜在MALDI样品靶,放入质谱仪用激光解析离解进行图像分析。
按上述方案,所述在动物组织切片图像分析中的应用为:首先将组织切片置于零下八十度下冷冻,再切成20微米厚度的切片,直接转移至半导体薄膜表面,固定半导体薄膜在MALDI样品靶,放入质谱仪后用激光解析离解进行图像分析。
按上述方案,所述在植物组织切片图像分析中的应用为:把半导体薄膜作为初膜,把植物组织切片放置于初膜表面,进一步施加压力,使植物组织切片填埋于半导体薄膜的纳米颗粒中后,得到含有植物组织切片的半导体薄膜,固定半导体薄膜在MALDI样品靶,放入质谱仪后用激光解析离解进行图像分析。本发明中,半导体颗粒的种类和用量依不同的样品而定,马弗炉高温灼烧后的半导体纳米颗粒需在玛瑙研钵磨细,使其分散均匀,以便使压制得到的半导体薄膜大小和厚度均匀。
本发明制备方法将半导体纳米颗粒材料在高压下压制制备均匀、大小厚度可控的薄膜,避免了现有技术中采用有机溶剂重结晶的不确定性,其获得的半导体薄膜能够吸收紫外光,在激光照射下处于价带的电子被激发到导带并发生隧穿,隧穿电子被组织切片或指纹中的中性分子俘获从而引发样品分子的电离和化学键断裂,由此进一步根据质谱信号成像。另外,采用本发明半导体薄膜所获得的谱图信号稳定,无背景干扰,信号强度与样品量之间成良好的线性关系,重现性好,灵敏度高,分辨率高。
本发明的有益效果如下:
(1)与现有MALDI质谱成像***相比,目前MALDI成像技术没有图像采集薄膜,一般是将有机小分子基质溶解于有机溶剂后以喷雾的形式覆盖组织切片,其由于有机基质与样品分子共结晶颗粒大小不一,容易导致质谱信号不稳定,定量关系差,分辨率低,并在低质量区产生大量背景干扰;而本发明利用半导体纳米材料的激光诱导隧道电子俘获原理使样品分子离子化,无背景干扰,克服了常规MALDI基质的局限性。
(2)本发明所述高分辨质谱成像***图像采集半导体薄膜采用半导体纳米颗粒在高压下压制成型即可得到,不仅方法简单,且获得的薄膜均匀、大小厚度可控,性质稳定,质谱信号稳定,表面均匀光滑,不产生背景干扰,可用于指纹分析和动植物组织切片分析,特别适合于小分子物质的准确质谱成像,便于质量控制和产业化。
附图说明
图1是实施例1所得的质谱图像,该图以己二烯雌酚分子离子峰成像,指纹按压在图像采集半导体薄膜上,激光扫描薄膜后得到质谱成像。
图2是实施例2所得拟南芥叶片的质谱图像,该图以茉莉酸分子离子峰成像。
图3是实施例3所得的小鼠脑质谱图像,该图以脑磷脂分子离子峰成像。
具体实施方式
为了更好地理解本发明,下面结合实施例进一步阐明本发明的内容,但本发明的内容不仅仅局限于下面的实施例。
实施例1
高分辨质谱成像***图像采集半导体薄膜的制备,该薄膜用于隐形指纹的成像分析,操作步骤依次如下:
1)用分析天平称取一定量的(Bi2O3)0.07(CoO)0.03(ZnO)0.9半导体纳米颗粒,比如10mg,材料的种类和量可据不同的样品而定;
2)将步骤1)得到的半导体纳米颗粒在350℃马弗炉中灼烧1小时,消除所吸附的有机分子的污染;
3)将步骤2)得到的半导体纳米颗粒进一步用玛瑙研钵磨细,使其分散均匀;
4)将步骤3)得到的半导体纳米粉末放入压片机的磨具,再放入压片机,施加4800kg压力,并在此压力下保持1分钟;
5)将步骤4)压制得半导体薄膜取出,保存在室温;
6)将指纹按压在步骤5)所得半导体薄膜表面,将薄膜固定于MALDI样品靶表面,放入质谱仪后用激光解析离解进行图像分析。
本实施例所得的质谱图像如图1所示,该图像是***己二烯雌酚的质谱图像。由图1可以看出,谱图信号稳定,无背景干扰,灵敏度高,分辨率高。
实施例2
高分辨质谱成像***图像采集半导体薄膜的制备,该薄膜用于植物激素茉莉酸的质谱成像,操作步骤如下:
1)用分析天平称取一定量的(Bi2O3)0.07(CoO)0.03(ZnO)0.9半导体纳米颗粒,比如10mg,材料的种类和量可据不同的样品而定;
2)将步骤1)得到的半导体纳米颗粒在350℃马弗炉中灼烧1小时,消除所吸附的有机分子的污染;
3)将步骤2)得到的半导体纳米颗粒进一步用玛瑙研钵磨细,使其分散均匀;
4)将步骤3)得到的半导体纳米粉末放入压片机的磨具,再放入压片机,施加2000kg压力,并在此压力下保持1分钟,得到半导体薄膜;
5)将步骤4)压制得半导体薄膜取出作为初膜,把拟南芥叶片放于初膜表面,再放入压片机,并将压力升至2000kg压力,并在此压力保持1分钟,得到含有叶片的半导体薄膜;
6)将步骤5)所得半导体薄膜固定于MALDI样品靶表面,放入质谱仪后用激光解析离解进行成像分析。
本实施例所得的质谱图像如图2所示,该图像是植物激素茉莉酸的质谱图像。由图2可以看出,谱图信号稳定,无背景干扰,灵敏度高,分辨率高。
实施例3
高分辨质谱成像***图像采集半导体薄膜的制备,该薄膜用于脑组织脑磷脂的质谱成像,操作步骤如下:
1)用分析天平称取一定量的(Bi2O3)0.07(CoO)0.03(ZnO)0.9半导体纳米颗粒,比如10mg,材料的种类和量可据不同的样品而定;
2)将步骤1)得到的半导体纳米颗粒在350℃马弗炉中灼烧1小时,消除所吸附的有机分子的污染;
3)将步骤2)得到的半导体纳米颗粒进一步用玛瑙研钵磨细,使其分散均匀;
4)将三分之二步骤3)得到的半导体纳米粉末放入压片机的磨具,再放入压片机,施加4800kg压力,并在此压力下保持1分钟,得到半导体薄膜;
5)将步骤4)压制得半导体薄膜取出,将小鼠脑于零下八十度冷冻后连续切片,每片厚度为20微米,直接将切片依次转移至薄膜表面;
6)将步骤5)所得薄膜固定于MALDI样品靶表面,放入质谱仪后用激光解析离解进行成像分析。
本实施例所得的质谱图像如图3所示,该图像是脑磷脂的质谱图像。由图3可以看出,谱图信号稳定,无背景干扰,灵敏度高,分辨率高。
显然,上述实施例仅仅是为清楚地说明所作的实例,而并非对实施方式的限制。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而因此所引申的显而易见的变化或变动仍处于本发明创造的保护范围之内。
Claims (10)
1.一种高分辨质谱成像***图像采集半导体薄膜,其特征在于,其是将半导体纳米颗粒灼烧去除表面附着的有机杂质后,再经研磨处理然后置于压片机中压制成膜得到的。
2.根据权利要求1所述的高分辨质谱成像***图像采集半导体薄膜,其特征在于,所述半导体纳米颗粒为(Bi2O3)0.07(CoO)0.03(ZnO)0.9半导体颗粒。
3.根据权利要求1所述的高分辨质谱成像***图像采集半导体薄膜,其特征在于,所述灼烧的温度为350℃,灼烧的时间为1小时。
4.权利要求1所述高分辨质谱成像***图像采集半导体薄膜的制备方法,其特征在于,包括如下步骤:
1)将半导体纳米颗粒在350℃马弗炉中灼烧1小时;
2)将步骤1)得到的半导体纳米颗粒进一步用玛瑙研钵磨细,使其分散均匀,得到半导体纳米粉末;
3)将步骤2)得到的半导体纳米粉末放入压片机的磨具,再放入压片机,施加压力压制得到半导体薄膜;
4)将步骤3)压制得半导体薄膜取出,室温保存。
5.根据权利要求4所述的制备方法,其特征在于,步骤3)所述压制为:2000kg~4800kg压力下压制1分钟。
6.权利要求1所述高分辨质谱成像***图像采集半导体薄膜在隐形指纹图像分析、动物组织切片图像分析、植物组织切片图像分析中的应用。
7.根据权利要求6所述的应用,其特征在于,所述应用为:将植物组织切片,动物组织切片或隐形指纹固定或按压在高分辨质谱成像***图像采集半导体薄膜上后,将半导体薄膜固定在样品靶上,直接放入质谱仪进行图像分析。
8.根据权利要求6~7任一所述的应用,其特征在于,所述隐形指纹图像分析的应用为:将隐形指纹直接按压于半导体薄膜表面后,固定半导体薄膜在MALDI样品靶,放入质谱仪用激光解析离解进行图像分析。
9.根据权利要求6~7任一所述的应用,其特征在于,所述动物组织切片图像分析的应用为:首先将动物组织切片置于零下八十度下冷冻,再切成20微米厚度的切片,直接转移至半导体薄膜表面,固定半导体薄膜在MALDI样品靶,放入质谱仪后用激光解析离解进行图像分析。
10.根据权利要求6~7任一所述的应用,其特征在于,所述植物组织切片图像分析的应用为:将半导体薄膜作为初膜,把植物组织切片放置于初膜表面,进一步施加压力,使组织切片填埋于半导体薄膜的纳米颗粒中后,得到含有植物组织切片的半导体薄膜,然后将其固定在MALDI样品靶,放入质谱仪后用激光解析离解进行图像分析。
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CN105973973A (zh) * | 2016-07-11 | 2016-09-28 | 华南师范大学 | 一种生物组织质谱成像方法 |
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