![用于细胞培养的显微镜解决方案 用于细胞培养的显微镜解决方案]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/laboratory-and-routine/clinical-laboratory/cell-culture/academic-laboratory-wallpaper.jpg/_jcr_content/renditions/original.image_file.100.100.0,0,1080,1080.file/academic-laboratory-wallpaper.jpg","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/laboratory-and-routine/clinical-laboratory/cell-culture/academic-laboratory-wallpaper.jpg/_jcr_content/renditions/original.image_file.360.360.0,0,1080,1080.file/academic-laboratory-wallpaper.jpg","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/laboratory-and-routine/clinical-laboratory/cell-culture/academic-laboratory-wallpaper.jpg/_jcr_content/renditions/original.image_file.768.768.0,0,1080,1080.file/academic-laboratory-wallpaper.jpg","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/laboratory-and-routine/clinical-laboratory/cell-culture/academic-laboratory-wallpaper.jpg/_jcr_content/renditions/original.image_file.1024.1024.0,0,1080,1080.file/academic-laboratory-wallpaper.jpg","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/laboratory-and-routine/clinical-laboratory/cell-culture/academic-laboratory-wallpaper.jpg/_jcr_content/renditions/original.image_file.1080.1080.0,0,1080,1080.file/academic-laboratory-wallpaper.jpg","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/laboratory-and-routine/clinical-laboratory/cell-culture/academic-laboratory-wallpaper.jpg/_jcr_content/renditions/original.image_file.1080.1080.0,0,1080,1080.file/academic-laboratory-wallpaper.jpg","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/laboratory-and-routine/clinical-laboratory/cell-culture/academic-laboratory-wallpaper.jpg/_jcr_content/renditions/original.image_file.1080.1080.0,0,1080,1080.file/academic-laboratory-wallpaper.jpg"})
用于细胞培养的显微镜解决方案
人工环境中生物体外培养基中细胞的生长
1934年,荷兰物理学家Frits Zernike提出了相差概念,从而彻底改变了活细胞的生物医学研究。蔡司在短短两年内就将Zernike的原始设计应用于第一台相差显微镜原型机中。该技术帮助Zernike赢得了1953年诺贝尔物理学奖;由于其非常适合用于未染色的样品(如玻片或塑料皿上的培养细胞),它至今仍是许多细胞生物学家的首选方法。
细胞培养研究在多个研究领域都十分重要,包括细胞生物学、制药、生物技术、细胞疗法和再生医学等。细胞培养又称组织培养,是指细胞在人工环境(生物体外)培养基中的生长,包括培养黏附细胞、悬浮细胞、原代细胞、干细胞、细菌、真菌或植物细胞。后三种有时会被更精确地称为微生物培养、真菌培养或植物组织培养。
模式生物和永生细胞系通常用于研究细胞或组织的生物学特性。此类细胞系会在培养皿、烧瓶或多孔板等特殊容器中进行培养。含有营养素和可选补充剂的培养基为优化细胞生长提供了必要条件。根据细胞类型的不同,必须维持特定的温度、湿度、二氧化碳和氧气水平,才能更好地模拟体内条件。大多数哺乳动物细胞系均在37° C和5%二氧化碳的培养装置中进行培养。
对显微镜的要求
细胞培养实验室每天使用显微镜来检查细胞生长或细胞增殖以及细胞活力,检查项目包括细胞融合度、细胞形态是否正常、是否存在污染以及何时需要更换培养基。而这些任务通常需要放大倍率可达50倍至200倍的相差显微镜。对此,您需要提高速度,缩短培养装置外的时间。因此,您的细胞培养显微镜应小巧精致,以便放置在超净工作台内或距离培养装置很近的实验台上。简单易用的显微镜周转快,可大幅减少对细胞的损伤。细胞融合达到一定水平后就需要对细胞进行转移,在将其转移到新的培养容器前,使用细胞计数器或计数室(如Makler室)确定细胞数量,然后计算出适当的稀释系数。良好的细胞培养实践至关重要,因为它为您在研究中获得有意义和可重复的结果奠定了基础。
在细胞培养实验室中还会进行许多其他显微镜操作。典型的试验包括划痕或伤口愈合试验、活死试验和细胞侵袭或易位试验。除了相差和明场观察,荧光的使用频率也很高,并逐渐成为了标准选择。细胞或组织中的蛋白质可以使用免疫荧光标记进行标记。DAPI、Hoechst、GFP、Alexa 488、RFP、Texas Red和Cy3等各种荧光基团可让您使用多通道荧光显微镜对信号进行区分和定位。
另外,活细胞可以(通过病毒或非病毒转染)转染外来DNA或RNA,以表达荧光蛋白等。如需稳定的转染,这一过程可能相当繁琐,因此荧光显微镜在此大有助益。表达水平和转染效率是该过程中的关键指标。使用发光二极管(LED)可以更好地实现低光毒性的荧光可视化和成像。与汞弧灯等其他荧光照明光源相比,紫外线(UV)产生的光毒性影响更小。此外,LED的使用寿命明显更长,而且无需维护。
应用实例
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