A novel and effective separation method for single mitochondria analysis

Pflugradt, René; Schmidt, Ulrike; Landenberger, Benjamin; Sänger, Timo; Lutz-Bonengel, Sabine

doi:10.1016/j.mito.2010.12.009

Cited by 19 publications

(14 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition to single cells including focal areas of fibrosis, inflammation and fatty infiltration, niches not easily studied by homogenate approaches. LCM of organelles such as mitochondria [11] open the possibilities of subcellular protein measurement by the LCM-ICP-QQQ method.…”

Section: Discussionmentioning

confidence: 99%

Single Cell Multiplex Protein Measurements through Rare Earth Element Immunolabeling, Laser Capture Microdissection and Inductively Coupled Mass Spectrometry

Liba

Wanagat

2014

J Cytol Histol

View full text Add to dashboard Cite

Complex diseases such as heart disease, stroke, cancer, and aging are the primary causes of death in the US. These diseases cause heterogeneous conditions among cells, conditions that cannot be measured in tissue homogenates and require single cell approaches. Understanding protein levels within tissues is currently assayed using various molecular biology techniques (e.g., Western blots) that rely on milligram to gram quantities of tissue homogenates or immunofluorescent (IF) techniques that are limited by spectral overlap. Tissue homogenate studies lack references to tissue structure and mask signals from individual or rare cellular events. Novel techniques are required to bring protein measurement sensitivity to the single cell level and offer spatiotemporal resolution and scalability. We are developing a novel approach to protein quantification by exploiting the inherently low concentration of rare earth elements (REE) in biological systems. By coupling REE-antibody immunolabeling of cells with laser capture microdissection (LCM) and ICP-QQQ, we are achieving multiplexed protein measurement in histological sections of single cells. This approach will add to evolving single cell techniques and our ability to understand cellular heterogeneity in complex biological systems and diseases.

show abstract

Section: Discussionmentioning

confidence: 99%

Single Cell Multiplex Protein Measurements through Rare Earth Element Immunolabeling, Laser Capture Microdissection and Inductively Coupled Mass Spectrometry

Liba

Wanagat

2014

J Cytol Histol

View full text Add to dashboard Cite

show abstract

“…When the intracellular contents spilled, the OT were switched on to capture the fluorescent mitochondria, which were finally directed toward a micropipette for collection and PCR analysis. Separation of mitochondria by OT has been proved a better approach as compared to laser capture microdissection and flow cytometry (Pflugradt et al, 2011).…”

Section: Genetic Materials Manipulationmentioning

confidence: 99%

Genetic Material Manipulation and Modification by Optical Trapping and Nanosurgery-A Perspective

Blázquez‐Castro

Fernández‐Piqueras

Santos

2020

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Light can be employed as a tool to alter and manipulate matter in many ways. An example has been the implementation of optical trapping, the so called optical tweezers, in which light can hold and move small objects with 3D control. Of interest for the Life Sciences and Biotechnology is the fact that biological objects in the size range from tens of nanometers to hundreds of microns can be precisely manipulated through this technology. In particular, it has been shown possible to optically trap and move genetic material (DNA and chromatin) using optical tweezers. Also, these biological entities can be severed, rearranged and reconstructed by the combined use of laser scissors and optical tweezers. In this review, the background, current state and future possibilities of optical tweezers and laser scissors to manipulate, rearrange and alter genetic material (DNA, chromatin and chromosomes) will be presented. Sources of undesirable effects by the optical procedure and measures to avoid them will be discussed. In addition, first tentative approaches at cellular-level genetic and organelle surgery, in which genetic material or DNA-carrying organelles are extracted out or introduced into cells, will be presented.

show abstract

“…Mitochondria from fresh murine liver tissue was prepared as described earlier. 22 Protoplasts from the root of Arabidopsis thaliana with a diameter of 40 mm were chosen for the experiments (for preparation see ref. 24).…”

Section: Cell Preparationmentioning

confidence: 99%

Microfluidic sorting of arbitrary cells with dynamic optical tweezers

et al. 2012

Self Cite

View full text Add to dashboard Cite

Optical gradient forces generated by fast steerable optical tweezers are highly effective for sorting small populations of cells in a lab-on-a-chip environment. The presented system can sort a broad range of different biological specimens by an automated optimisation of the tweezer path and velocity profile. The optimal grab positions for subsequent trap and cell displacements are estimated from the intensity of the bright field image, which is derived theoretically and proven experimentally. We exhibit rapid displacements of 2 μm small mitochondria, yeast cells, rod-shaped bacteria and 30 μm large protoplasts. Reliable sorting of yeast cells in a microfluidic chamber by both morphological criteria and by fluorescence emission is demonstrated.

show abstract

A novel and effective separation method for single mitochondria analysis

Cited by 19 publications

References 45 publications

Single Cell Multiplex Protein Measurements through Rare Earth Element Immunolabeling, Laser Capture Microdissection and Inductively Coupled Mass Spectrometry

Single Cell Multiplex Protein Measurements through Rare Earth Element Immunolabeling, Laser Capture Microdissection and Inductively Coupled Mass Spectrometry

Genetic Material Manipulation and Modification by Optical Trapping and Nanosurgery-A Perspective

Microfluidic sorting of arbitrary cells with dynamic optical tweezers

Contact Info

Product

Resources

About