Riglar David T, Baum Jake
Division of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research, Melbourne, VIC, Australia.
Methods Mol Biol. 2013;923:269-80. doi: 10.1007/978-1-62703-026-7_18.
Cellular imaging has reemerged in recent years as a powerful approach to provide researchers with a direct measure of essential molecular events in a cell's life, ranging in scale from broad morphological observations of whole cells to intricate single molecule imaging. When combined with quantitative image analysis, the available imaging techniques can act as a critical means to confirm hypotheses, drive the formation of new theories or provide accurate determination of protein localization at subcellular and nanometer scales. Here, we describe two methodological approaches for imaging the transient step of malaria parasite invasion of the human erythrocyte. When applied to image the most virulent human malaria parasite, Plasmodium falciparum, the first approach, using live time-lapse wide-field microscopy, allows the capture of transient events during invasion and postinvasion intra-erythrocytic development, while the second, using immunofluorescence assay (IFA) of fixed samples, allows high-definition exploration of parasite architecture on multiple platforms.
近年来，细胞成像作为一种强大的方法重新出现，为研究人员提供了对细胞生命中基本分子事件的直接测量，其规模从对整个细胞的广泛形态观察到复杂的单分子成像。当与定量图像分析相结合时，现有的成像技术可以作为一种关键手段，用于证实假设、推动新理论的形成或在亚细胞和纳米尺度上准确确定蛋白质的定位。在这里，我们描述了两种用于对疟原虫入侵人类红细胞的瞬态步骤进行成像的方法。当应用于对最具毒性的人类疟原虫恶性疟原虫进行成像时，第一种方法是使用实时延时宽场显微镜，它可以在入侵和入侵后红细胞内发育过程中捕捉瞬态事件，而第二种方法是对固定样本进行免疫荧光测定（IFA），它可以在多个平台上对寄生虫结构进行高清探索。
