Angular Light-Scattering Studies on Isolated Mitochondria

Gotterer, Gerald S.; Thompson, T. E.; Lehninger, Albert L.

doi:10.1083/jcb.10.1.15

Cited by 34 publications

(7 citation statements)

References 26 publications

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“…For example, inside a cell, backscattering is reduced as the relative refractive index of intracellular organelles (i.e., with respect to that of the cytoplasm) barely reaches n rel = 1.1 (e.g., lipid droplet refractive index n LD = 1.48 − 1.53 43 , cytoplasm refractive index n c = 1.36 − 1.375 44 ), whereas for glass microcylinders in water, n rel = 1.17. Angular light-scattering studies show that even in organelles with complex internal structure, such as mitochondria, scattering predominantly occurs in the forward direction 45 .…”

Section: Discussionmentioning

confidence: 99%

Extending calibration-free force measurements to optically-trapped rod-shaped samples

Català

Marsà

Montes-Usategui

et al. 2017

Sci Rep

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Optical trapping has become an optimal choice for biological research at the microscale due to its non-invasive performance and accessibility for quantitative studies, especially on the forces involved in biological processes. However, reliable force measurements depend on the calibration of the optical traps, which is different for each experiment and hence requires high control of the local variables, especially of the trapped object geometry. Many biological samples have an elongated, rod-like shape, such as chromosomes, intracellular organelles (e.g., peroxisomes), membrane tubules, certain microalgae, and a wide variety of bacteria and parasites. This type of samples often requires several optical traps to stabilize and orient them in the correct spatial direction, making it more difficult to determine the total force applied. Here, we manipulate glass microcylinders with holographic optical tweezers and show the accurate measurement of drag forces by calibration-free direct detection of beam momentum. The agreement between our results and slender-body hydrodynamic theoretical calculations indicates potential for this force-sensing method in studying protracted, rod-shaped specimens.

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Section: Discussionmentioning

confidence: 99%

Extending calibration-free force measurements to optically-trapped rod-shaped samples

Català

Marsà

Montes-Usategui

et al. 2017

Sci Rep

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“…Upon opening of the mPT pore, swelling of the mitochondria is an invariable event. Thus, the most direct way of testing for mPT in isolated mitochondria is by following mitochondrial light scatter (Gotterer et al, 1961), while for in situ mitochondria, time-lapse of microscopic visualization of mitotargeted fluorophores is warranted (Doczi et al, 2011), (Doczi et al, 2016). In addition, the method of quenching mitochondrially-trapped calcein by exogenously added cobalt is also widely used (Petronilli et al, 1998), however, the materials are too toxic for some cell types (Doczi et al, 2016).…”

Section: Bioenergetic Pitfalls In Search Of the Mptmentioning

confidence: 99%

Mitochondrial permeability transition pore: Back to the drawing board

Chinopoulos

2018

Neurochemistry International

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Current models theorizing on what the mitochondrial permeability transition (mPT) pore is made of, implicate the c-subunit rings of ATP synthase complex. However, two very recent studies, one on atomistic simulations and in the other disrupting all genes coding for the c subunit disproved those models. As a consequence of this, the structural elements of the pore remain unknown. The purpose of the present short-review is to (i) briefly review the latest findings, (ii) serve as an index for more comprehensive reviews regarding mPT specifics, (iii) reiterate on the potential pitfalls while investigating mPT in conjunction to bioenergetics, and most importantly (iv) suggest to those in search of mPT pore identity, to also look elsewhere.

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“…This is beneficial in the study of expansion/contraction or structural changes of, for instance, mitochondria [27,36] or chloroplasts, which is traditionally done with light scattering [37].…”

Section: Forward and Backward Thg From Multilayermentioning

confidence: 99%

Numerical second- and third-harmonic generation microscopy

et al. 2013

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A full numerical description of second-and third-harmonic generation (SHG and THG) at the focus of a nonlinear microscope is presented. The numerical implementation takes into account reflections and refraction by an arbitrary number of interfaces perpendicular to the optical axis in the focal region. The calculation of the second-and third-harmonic far-field radiation pattern is based on a Green function approach and is presented for any collection direction. The calculations are sped up by using the chirp-z transform for the focusing fields as well as for the far-field radiation calculation. Numerical evidence is presented for deviations in the measurement of the secondorder nonlinear susceptibility ratio ρ ≡ χ 2 yyy ∕χ 2 yxx of collagen fibers in SHG microscopy at high excitation numerical aperture. When interface reflections are taken into account, significant direct backward THG is demonstrated from interfaces and multilayer structures.

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Angular Light-Scattering Studies on Isolated Mitochondria

Cited by 34 publications

References 26 publications

Extending calibration-free force measurements to optically-trapped rod-shaped samples

Extending calibration-free force measurements to optically-trapped rod-shaped samples

Mitochondrial permeability transition pore: Back to the drawing board

Numerical second- and third-harmonic generation microscopy

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