A Modified Magnified Analysis of Proteome (MAP) Method for Super-Resolution Cell Imaging that Retains Fluorescence

被引:9
作者
Woo, Jiwon [1 ,2 ]
Seo, Jeong-Min [3 ,4 ,5 ]
Lee, Mirae [1 ,2 ]
Kim, Juyoung [1 ]
Min, Sol [3 ,4 ]
Kim, Sang-Tae [6 ]
Ku, Seockmo [7 ]
Park, Jeong-Yoon [1 ,2 ]
机构
[1] Yonsei Univ, Gangnam Severance Hosp, Spine & Spinal Cord Inst, Dept Neurosurg,Coll Med, Seoul 06273, South Korea
[2] Yonsei Univ, Brain Korea 21 PLUS Project Med Sci, Seoul 03722, South Korea
[3] Seoul Natl Univ, Sch Dent, Cellular Reprogramming & Embryo Biotechnol Lab, Seoul 08826, South Korea
[4] Seoul Natl Univ, Sch Dent, Dent Res Inst, Seoul 08826, South Korea
[5] NeoRegen Biotech Co Ltd, Biomed Res Inst, Gyeonggi Do 16614, South Korea
[6] Seoul Natl Univ, Dept Neurol, Bundang Hosp, Seongnam Si 13605, Gyeonggi Do, South Korea
[7] Middle Tennessee State Univ, Coll Basic & Appl Sci, Sch Agr, Fermentat Sci Program, Murfreesboro, TN 37132 USA
基金
新加坡国家研究基金会;
关键词
STRUCTURED-ILLUMINATION MICROSCOPY; EXPANSION MICROSCOPY; RESOLUTION LIMIT; LOCALIZATION; BREAKING; MIR-671; CANCER;
D O I
10.1038/s41598-020-61156-2
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Biological systems consist of a variety of distinct cell types that form functional networks. Super-resolution imaging of individual cells is required for better understanding of these complex systems. Direct visualization of 3D subcellular and nano-scale structures in cells is helpful for the interpretation of biological interactions and system-level responses. Here we introduce a modified magnified analysis of proteome (MAP) method for cell super-resolution imaging (Cell-MAP) which preserves cell fluorescence. Cell-MAP expands cells more than four-fold while preserving their overall architecture and three-dimensional proteome organization after hydrogel embedding. In addition, Optimized-Cell-MAP completely preserves fluorescence and successfully allows for the observation of tagged small molecular probes containing peptides and microRNAs. Optimized-Cell-MAP further successfully applies to the study of structural characteristics and the identification of small molecules and organelles in mammalian cells. These results may give rise to many other applications related to the structural and molecular analysis of smaller assembled biological systems.
引用
收藏
页数:14
相关论文
共 33 条
[1]   Biological Insight from Super-Resolution Microscopy: What We Can Learn from Localization-Based Images [J].
Baddeley, David ;
Bewersdorf, Joerg .
ANNUAL REVIEW OF BIOCHEMISTRY, VOL 87, 2018, 87 :965-989
[2]   In vitro quantification of specific microRNA using molecular beacons [J].
Baker, Meredith B. ;
Bao, Gang ;
Searles, Charles D. .
NUCLEIC ACIDS RESEARCH, 2012, 40 (02) :e13
[3]   Imaging intracellular fluorescent proteins at nanometer resolution [J].
Betzig, Eric ;
Patterson, George H. ;
Sougrat, Rachid ;
Lindwasser, O. Wolf ;
Olenych, Scott ;
Bonifacino, Juan S. ;
Davidson, Michael W. ;
Lippincott-Schwartz, Jennifer ;
Hess, Harald F. .
SCIENCE, 2006, 313 (5793) :1642-1645
[4]   Expansion microscopy [J].
Chen, Fei ;
Tillberg, Paul W. ;
Boyden, Edward S. .
SCIENCE, 2015, 347 (6221) :543-548
[5]  
Chozinski TJ, 2016, NAT METHODS, V13, P485, DOI [10.1038/NMETH.3833, 10.1038/nmeth.3833]
[6]   Practical limits of resolution in confocal and non-linear microscopy [J].
Cox, G ;
Sheppard, CJR .
MICROSCOPY RESEARCH AND TECHNIQUE, 2004, 63 (01) :18-22
[7]   Imaging cellular ultrastructures using expansion microscopy (U-ExM) [J].
Gambarotto, Davide ;
Zwettler, Fabian U. ;
Le Guennec, Maeva ;
Schmidt-Cernohorska, Marketa ;
Fortun, Denis ;
Borgers, Susanne ;
Heine, Joern ;
Schloetel, Jan-Gero ;
Reuss, Matthias ;
Unser, Michael ;
Boyden, Edward S. ;
Sauer, Markus ;
Hamel, Virginie ;
Guichard, Paul .
NATURE METHODS, 2019, 16 (01) :71-+
[8]  
Heintzmann Rainer, 2006, Briefings in Functional Genomics & Proteomics, V5, P289, DOI 10.1093/bfgp/ell036
[9]   BREAKING THE DIFFRACTION RESOLUTION LIMIT BY STIMULATED-EMISSION - STIMULATED-EMISSION-DEPLETION FLUORESCENCE MICROSCOPY [J].
HELL, SW ;
WICHMANN, J .
OPTICS LETTERS, 1994, 19 (11) :780-782
[10]   Recent advancements in structured-illumination microscopy toward live-cell imaging [J].
Hirano, Yasuhiro ;
Matsuda, Atsushi ;
Hiraoka, Yasushi .
MICROSCOPY, 2015, 64 (04) :237-249