Imaging flow cytometry analysis of intracellular pathogens

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• 作者：Viraga Haridas^a ; ^b ; Shahin Ranjbar^a ; ^b ; Ivan A. Vorobjev^c ; ^d ; ^e ; Anne E. Goldfeld^a ; ^b ; ^{Anne.Goldfeld@childrens.harvard.edu} ; Natasha S. Barteneva^a ; ^b ; ^c ; ^{natasha.barteneva@childrens.harvard.edu}
• 关键词：An5 ; annexin V ; APC ; antigen-presenting cells ; BDS ; Bright Detail Similarity ; BF ; bright field ; CFSE ; carboxyfluoresceinsuccinimidyl ester ; CFU ; colony-forming unit ; CMDFA ; 5-chloromethylfluoresceinDiacetate ; DAPI ; (4&prime ; 6-diamidino-2-phenylindole) ; DHR ; dihydrorhodamine ; DMEM ; Dulbecco&rsquo ; s modified Eagle medium ; FP ; fluorescent protein ; EGFP ; enhanced green fluorescent protein ; em. ; emission ; EVs ; extracellular vesicles ; EYFP ; enhanced yellow fluorescent protein ; ex. ; excitation ; H2DCFD
• 刊名：Methods
• 出版年：2017
• 出版时间：1 January 2017
• 年：2017
• 卷：112
• 期：Complete
• 页码：91-104
• 全文大小：2688 K
• 卷排序：112

文摘

Imaging flow cytometry (IFC) characterize hundreds thousands of cellular images using hundreds of measurements of morphological and fluorescence cellular features. Complex morphology of host-intracellular parasite interaction can be quantified in statistically robust manner using IFC. Feature Finder algorithm is useful in defining statistically significant differences in morphological and fluorescent features of cells. IFC revealed that M. tuberculosis internalized bacilli were co-localized to a greater degree at late endosome/lysosome CD107 and late endosome Rab7 compartment than Rab5 early endosome compartment.