The effects of refractive index heterogeneity within kidney tissue on multiphoton fluorescence excitation microscopy

Young, P. G.; Clendenon, Sherry G.; Byars, Jason M.; Dunn, Kenneth W.

doi:10.1111/j.1365-2818.2010.03448.x

Cited by 23 publications

(19 citation statements)

References 37 publications

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“…Residual differences in refractive index lead to scattering and aberrations that degrade the point-spread function at depth. Axial resolution is affected most strongly, particularly when using high numerical aperture objectives, reducing clarity of optical sections and three dimensional reconstructions [8]. The alternative clearing method known as Clarity that employs hydrogel scaffolds and electrophoresis to remove lipids [9] affords excellent index matching, but is too labor intensive and tissue altering for practical clinical histology.…”

Section: Introductionmentioning

confidence: 99%

Multiphoton microscopy with clearing for three dimensional histology of kidney biopsies

Olson¹,

Levene²,

Torres³

2016

Biomed. Opt. Express

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We present a multiphoton microscopy approach with clearing optimized for pathology evaluation producing image quality comparable to traditional histology. Use of benzyl alcohol/benzyl benzoate with 4',6-diamidino-2-phenylindole and eosin in an optimized imaging setup results in optical sections nearly indistinguishable from traditionally-cut sections. Application to human renal tissue demonstrates diagnostic-level image quality can be maintained through 1 millimeter of tissue. Three dimensional perspectives reveal changes of glomerular capsule cells not evident on single sections. Collagen-derived second harmonic generation can be visualized through entire biopsies. Multiphoton microscopy with clearing has potential for increasing the yield of histologic evaluation of biopsy specimens.OCIS codes: (180.0180) Microscopy; (180.4315) Nonlinear microscopy; (180.6900) Three-dimensional microscopy.

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Section: Introductionmentioning

confidence: 99%

Multiphoton microscopy with clearing for three dimensional histology of kidney biopsies

Olson¹,

Levene²,

Torres³

2016

Biomed. Opt. Express

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“…As for the “optical transparency” of a given organ, it strongly depends on its homogeneity. Indeed, light undergoes scattering when crossing the interface between component of the tissues with different refractive indexes 38 , 39 . It is also generally true that highly vascularized tissues are less optically transparent due to the light absorption and scattering of the red blood cells.…”

Section: Imaging Modality and Depthmentioning

confidence: 99%

“…Although a rigorous side by side comparison of the various organs has never been performed, it has been empirically determined that tissues such as the brain are optically more suitable for IVM than skeletal muscle, skin, liver or kidney. Notably, in the last couple of years several approaches have been used to “clear” tissues by using special solutions that either reduce the difference between the refractive index in the various tissue layers 38 , 39 or make them optically transparent 40 . This approach has enabled imaging neurons labeled with genetically encoded fluorescent proteins up to 8 mm below the surface with an unprecedented resolution (Fig.…”

Section: Imaging Modality and Depthmentioning

confidence: 99%

Intravital microscopy

et al. 2012

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Intravital microscopy is an extremely powerful tool that enables imaging several biological processes in live animals. Recently, the ability to image subcellular structures in several organs combined with the development of sophisticated genetic tools has made possible extending this approach to investigate several aspects of cell biology. Here we provide a general overview of intravital microscopy with the goal of highlighting its potential and challenges. Specifically, this review is geared toward researchers that are new to intravital microscopy and focuses on practical aspects of carrying out imaging in live animals. Here we share the know-how that comes from first-hand experience, including topics such as choosing the right imaging platform and modality, surgery and stabilization techniques, anesthesia and temperature control. Moreover, we highlight some of the approaches that facilitate subcellular imaging in live animals by providing numerous examples of imaging selected organelles and the actin cytoskeleton in multiple organs.

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“…A further advantage of using near-infrared wavelengths, instead of ultraviolet light, for fluorescence excitation is the reduction of scattering and increase in imaging penetration depth in the sample from approximately 20 µm to 500-600 µm. However, in the kidney, the maximal penetration depth is approximately 150 µm owing to substantial scattering of emission photons from the high refractory index and heterogeneity of kidney tissue (Young et al, 2011a;Young et al, 2011b). A useful application of FLIM in conjunction with MPM in the kidney is measuring the fluorescence lifetime of NAD(P)H. NAD(P)H lifetime measurements are widely used for metabolic and redox imaging in vitro and in vivo (Bird et al, 2005;Skala et al, 2007a;Skala et al, 2007b;Leite-Silva et al, 2013;Thorling et al, 2013).…”

Section: Figure 51: Acute Kidney Injury and Chronic Kidney Disease Amentioning

confidence: 99%

mentioning

confidence: 99%

Identification of potential mediators of oxidative damage and targets for novel treatments for chronic kidney disease

Small¹

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Chronic kidney disease (CKD) is a common and serious problem causing a significant burden to healthcare systems and, most importantly, the affected individual. Despite this, few successful treatments exist for CKD. Understanding the pathophysiological features that are common to chronic kidney pathologies is vital to identify targets for treatment. Oxidative stress has been implicated in all chronic diseases, acting via highly conserved pathways of disease progression.However, evidence to date shows limited benefit of antioxidants as therapies for CKD patients, suggesting a need to better identify the pathways that are stimulated.This thesis focuses on the role of oxidative stress, the regulation of mitochondrial homeostasis, the action of antioxidant compounds, and the disruption of oxidant signalling networks in chronic kidney pathologies (reviewed in Chapter 1). The aims of this project were to: (1) to use an in vitro model of oxidative stress-induced kidney injury to determine how a failure in balance between oxidative stress and oxidant control causes the cellular characteristics of CKD, then to determine whether exogenous antioxidants can prevent and restore renal cell bioenergetics and reduce cell injury; (2) to use an in vivo model of oxidative stress-induced kidney injury with molecular and metabolic imaging to measure potential biomarkers identified in Aim 1, in association with changes in kidney structure and function, and determine whether treatment with an antioxidant therapy modulates kidney disease pathology; and (3) to explore the links between biomarkers of oxidative stress and clinical characteristics of CKD in patients to determine whether a lifestyle intervention in conjunction with standard nephrology care improve systemic oxidative stress and whether this influences kidney function. The renal specific in vitro, in vivo, and clinical models and the methods used are explained in Chapter 2.Chapter 3 reports the separate and cumulative effects of oxidative stress, mitochondrial dysfunction and cell senescence in promoting loss of renal cells in an in vitro model of kidney disease. The results demonstrate that oxidative stress and cell senescence cause mitochondrial destabilization and kidney tubular epithelial cell loss. This may contribute to the development of cellular and tissue atrophy seen in CKD.Chapter 4 further defines the role of mitochondrial alterations as a consequence of a specificallyimpaired oxidant signalling regulatory mechanism, namely, peroxisome proliferator-activated receptor-gamma (PPARγ)-regulated mitochondrial biogenesis. Oxidative stress promoted mitochondrial destabilisation in kidney proximal tubular epithelial cells, in association with ii increased PPARγ Serine 112 phosphorylation. Despite their positive effects in other tissues, PPARγ agonists failed to protect proximal tubular epithelial (PTE) cells, and appear to be detrimental to kidney PTE cell health when oxidative stress induces the cell damage.Chapter 5 examined mediators of oxidative stress in an in...

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The effects of refractive index heterogeneity within kidney tissue on multiphoton fluorescence excitation microscopy

Cited by 23 publications

References 37 publications

Multiphoton microscopy with clearing for three dimensional histology of kidney biopsies

Multiphoton microscopy with clearing for three dimensional histology of kidney biopsies

Intravital microscopy

Identification of potential mediators of oxidative damage and targets for novel treatments for chronic kidney disease

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