A Single-Cell Resolution Imaging Protocol of Mitochondrial DNA Dynamics in Physiopathology, mTRIP, Which Also Evaluates Sublethal Cytotoxicity

2016 ◽  
pp. 49-65 ◽  
Author(s):  
Laurent Chatre ◽  
Benjamin Montagne ◽  
Miria Ricchetti
Keyword(s):  
Mitochondrial Dna ◽  
Single Cell ◽  
Dna Dynamics ◽  
Imaging Protocol ◽  
Resolution Imaging

scholarly journals μCB-seq: Microfluidic cell barcoding and sequencing for high-resolution imaging and sequencing of single cells

2020 ◽  
Author(s):  
Tyler N. Chen ◽  
Anushka Gupta ◽  
Mansi Zalavadia ◽  
Aaron M. Streets
Keyword(s):  
High Resolution ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Protein Localization ◽  
Single Cells ◽  
Optical Measurements ◽  
Detection Sensitivity ◽  
High Resolution Imaging ◽  
Cell Analysis ◽  
Resolution Imaging

AbstractSingle-cell RNA sequencing (scRNA-seq) enables the investigation of complex biological processes in multicellular organisms with high resolution. However, many phenotypic features that are critical to understanding the functional role of cells in a heterogeneous tissue or organ are not directly encoded in the genome and therefore cannot be profiled with scRNA-seq. Quantitative optical microscopy has long been a powerful approach for characterizing diverse cellular phenotypes including cell morphology, protein localization, and chemical composition. Combining scRNA-seq with optical imaging has the potential to provide comprehensive single-cell analysis, allowing for functional integration of gene expression profiling and cell-state characterization. However, it is difficult to track single cells through both measurements; therefore, coupling current scRNA-seq protocols with optical measurements remains a challenge. Here, we report Microfluidic Cell Barcoding and Sequencing (μCB-seq), a microfluidic platform that combines high-resolution imaging and sequencing of single cells. μCB-seq is enabled by a novel fabrication method that preloads primers with known barcode sequences inside addressable reaction chambers of a microfluidic device. In addition to enabling multi-modal single-cell analysis, μCB-seq improves gene detection sensitivity, providing a scalable and accurate method for information-rich characterization of single cells.

scholarly journals 3P325 Direct quantification of mitochondria and mitochondrial DNA dynamics in living cells(Bioimaging,The 48th Annual Meeting of the Biophysical Society of Japan)

2010 ◽  
Vol 50 (supplement2) ◽  
pp. S202
Author(s):  
Yasutomo Nomura ◽  
Tomomi Nabeya ◽  
Kyohei Nakayama ◽  
Kazuki Nishimoto ◽  
Yui Januma ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Annual Meeting ◽  
Living Cells ◽  
Dna Dynamics ◽  
Biophysical Society ◽  
Direct Quantification

scholarly journals Identification of unique and shared mitochondrial DNA mutations in neurodegeneration and cancer by single-cell mitochondrial DNA structural variation sequencing (MitoSV-seq)

EBioMedicine ◽  
2020 ◽  
Vol 57 ◽  
pp. 102868
Author(s):  
Elham Jaberi ◽  
Emilie Tresse ◽  
Kirsten Grønbæk ◽  
Joachim Weischenfeldt ◽  
Shohreh Issazadeh-Navikas
Keyword(s):  
Mitochondrial Dna ◽  
Single Cell ◽  
Structural Variation ◽  
Mitochondrial Dna Mutations ◽  
Dna Mutations

scholarly journals Super‐Resolution Imaging and Optomechanical Manipulation Using Optical Nanojet for Nondestructive Single‐Cell Research

2021 ◽  
pp. 2100233
Author(s):  
Alina Karabchevsky ◽  
Tal Elbaz ◽  
Aviad Katiyi ◽  
Ofer Prager ◽  
Alon Friedman
Keyword(s):  
Single Cell ◽  
Super Resolution ◽  
Resolution Imaging

scholarly journals Heteroplasmy variability in individuals with biparentally inherited mitochondrial DNA

2020 ◽  
Author(s):  
Jesse Slone ◽  
Weiwei Zou ◽  
Shiyu Luo ◽  
Eric S Schmitt ◽  
Stella Maris Chen ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Single Cell ◽  
Strong Evidence ◽  
Genetic Factors ◽  
Single Cell Level ◽  
Biparental Inheritance ◽  
Cell Level ◽  
Single Population ◽  
Parent Of Origin ◽  
Two Populations

ABSTRACTWith very few exceptions, mitochondrial DNA (mtDNA) in humans is transmitted exclusively from mothers to their offspring, suggesting the presence of a strong evolutionary pressure favoring the exclusion of paternal mtDNA. We have recently shown strong evidence of paternal mtDNA transmission. In these rare situations, males exhibiting biparental mtDNA appear to be limited to transmitting just one of the mtDNA species to their offspring, while females possessing biparental mtDNA populations consistently transmit both populations to their offspring at a very similar heteroplasmy level. The precise biological and genetic factors underlying this unusual transmission event remain unclear. Here, we have examined heteroplasmy levels in various tissues among individuals with biparental inheritance. Our results indicate that individuals with biparental mtDNA have remarkable inter-tissue variability in heteroplasmy level. At the single-cell level, paternal mtDNA heteroplasmy in sperm varies dramatically, and many sperm possess only one of the two mtDNA populations originally in question. These results show a fundamental, parent-of-origin difference in how mtDNA molecules transmit and propagate. This helps explain how a single population of mtDNAs are transmitted from a father possessing two populations of mtDNA molecules, suggesting that some mtDNA populations may be favored over others when transmitted from the father.

scholarly journals Understanding mitochondrial DNA maintenance disorders at the single muscle fibre level

2019 ◽  
Vol 47 (14) ◽  
pp. 7430-7443 ◽  
Author(s):  
Diana Lehmann ◽  
Helen A L Tuppen ◽  
Georgia E Campbell ◽  
Charlotte L Alston ◽  
Conor Lawless ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Single Cell ◽  
Respiratory Chain ◽  
Genetic Defect ◽  
Clear Correlation ◽  
Muscle Fibres ◽  
Single Muscle Fibre ◽  
Respiratory Chain Deficiency ◽  
Mtdna Deletions ◽  
Mtdna Deletion

Abstract Clonal expansion of mitochondrial DNA (mtDNA) deletions is an important pathological mechanism in adults with mtDNA maintenance disorders, leading to a mosaic mitochondrial respiratory chain deficiency in skeletal muscle. This study had two aims: (i) to determine if different Mendelian mtDNA maintenance disorders showed similar pattern of mtDNA deletions and respiratory chain deficiency and (ii) to investigate the correlation between the mitochondrial genetic defect and corresponding respiratory chain deficiency. We performed a quantitative analysis of respiratory chain deficiency, at a single cell level, in a cohort of patients with mutations in mtDNA maintenance genes. Using the same tissue section, we performed laser microdissection and single cell genetic analysis to investigate the relationship between mtDNA deletion characteristics and the respiratory chain deficiency. The pattern of respiratory chain deficiency is similar with different genetic defects. We demonstrate a clear correlation between the level of mtDNA deletion and extent of respiratory chain deficiency within a single cell. Long-range and single molecule PCR shows the presence of multiple mtDNA deletions in approximately one-third of all muscle fibres. We did not detect evidence of a replicative advantage for smaller mtDNA molecules in the majority of fibres, but further analysis is needed to provide conclusive evidence.

Live-Cell Probe for In Situ Single-Cell Monitoring of Mitochondrial DNA Methylation

ACS Sensors ◽  
2021 ◽  
Author(s):  
Han Zhao ◽  
Donghan Ma ◽  
Junkai Xie ◽  
Oscar Sanchez ◽  
Fang Huang ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Mitochondrial Dna ◽  
Single Cell ◽  
Live Cell ◽  
Cell Monitoring
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