High-resolution visualization of mouse cardiac microvasculature using optical histology

Moy, Austin J.; Lo, Patrick; Choi, Bernard

doi:10.1364/boe.5.000069

Cited by 15 publications

(25 citation statements)

References 43 publications

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“…11,12 Briefly, this method first involved cardiac perfusion of the lipophilic fluorescent dye DiI to label the mouse microvasculature, followed by tissue harvest and preparation similar to the procedure previously described. All procedures were approved by the Institutional Animal Care and Use Committee at University of California, Irvine.…”

Section: Optical Histologymentioning

confidence: 99%

“…The result is that both techniques are more challenging to implement than simply immersing the tissue in an optical clearing agent. We recently described a method that we call optical histology, 11,12 which involves applying optical clearing using FocusClear to thick (∼1 mm) tissue sections and subsequent imaging with confocal fluorescence microscopy. With optical histology, we demonstrated high-resolution visualization of fluorescent structures up to depths of 500 to 700 μm within each section.…”

Section: Introductionmentioning

confidence: 99%

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Optical properties of mouse brain tissue after optical clearing with FocusClear™

et al. 2015

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Abstract. Fluorescence microscopy is commonly used to investigate disease progression in biological tissues. Biological tissues, however, are strongly scattering in the visible wavelengths, limiting the application of fluorescence microscopy to superficial (<200 μm) regions. Optical clearing, which involves incubation of the tissue in a chemical bath, reduces the optical scattering in tissue, resulting in increased tissue transparency and optical imaging depth. The goal of this study was to determine the time-and wavelength-resolved dynamics of the optical scattering properties of rodent brain after optical clearing with FocusClear™. Light transmittance and reflectance of 1-mm mouse brain sections were measured using an integrating sphere before and after optical clearing and the inverse adding doubling algorithm used to determine tissue optical scattering. The degree of optical clearing was quantified by calculating the optical clearing potential (OCP), and the effects of differing OCP were demonstrated using the optical histology method, which combines tissue optical clearing with optical imaging to visualize the microvasculature. We observed increased tissue transparency with longer optical clearing time and an analogous increase in OCP. Furthermore, OCP did not vary substantially between 400 and 1000 nm for increasing optical clearing durations, suggesting that optical histology can improve ex vivo visualization of several fluorescent probes.

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Section: Optical Histologymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optical properties of mouse brain tissue after optical clearing with FocusClear™

et al. 2015

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“…Despite advancements in the field, our understanding of the adult heart structure and its remodeling following disease is still limited by the inability to provide three-dimensional (3D) images of the myocardium at cellular resolution. There is a growing trend to study the 3D structure of organs and tissues, requiring researchers to work with volumes rather than thin sections [3]. Confocal microscopy is a well-established imaging technique that plays an important role in studying tissue with high magnification.…”

Section: Introductionmentioning

confidence: 99%

3D imaging in CUBIC-cleared mouse heart tissue: going deeper

Nehrhoff¹,

Bocancea²,

Vaquero³

et al. 2016

Biomed. Opt. Express

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Abstract:The ability to acquire high resolution 3D images of the heart enables to study heart diseases more in detail. In this work, the CUBIC (clear, unobstructed brain imaging cocktails and computational analysis) clearing protocol was optimized for thick mouse heart sections to enhance the penetration depth of the confocal microscope lasers into the tissue. In addition, the optimized CUBIC clearing of the heart enhances antibody penetration into the tissue by a factor of five. The present protocol enables deep 3D high-quality image acquisition in the heart allowing a much more accurate assessment of the cellular and structural changes that underlie heart diseases. Sadayappan, "Cardiac muscle organization revealed in 3-D by imaging whole-mount mouse hearts using twophoton fluorescence and confocal microscopy,"

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“…For the last decade, optical clearing technique to improve the imaging depth has been actively investigated [21][22][23][24][25][26][27][28][29][30][31][32][33][34]. As the mismatch in refractive index (RI) between cellular components in tissue is the major cause of light scattering, selective removal of cellular components with relatively low or high refractive index can reduce the light scattering and make the tissue more optically transparent.…”

Section: Introductionmentioning

confidence: 99%

“…The most frequently used approach is replacing interstitial fluids (RI~1.33) in inter-and intra-cellular space with optical clearing agents (OCAs) which have refractive indices of ~1.46, matched to the average refractive index of most biological samples. It has been demonstrated that several OCAs such as glycerol [26- [30][31][32] and BABB [33] can enhance imaging depth in various organs such as brain [22][23][24][25][26], skin [27][28][29], pancreas [30], colon [31] and heart [32][33][34]. However, optical clearing effect of OCAs on LN has not yet been clearly demonstrated.…”

Section: Introductionmentioning

confidence: 99%

Optical clearing based cellular-level 3D visualization of intact lymph node cortex

Song

Seo

Choe

et al. 2015

Biomed. Opt. Express

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Lymph node (LN) is an important immune organ that controls adaptive immune responses against foreign pathogens and abnormal cells. To facilitate efficient immune function, LN has highly organized 3D cellular structures, vascular and lymphatic system. Unfortunately, conventional histological analysis relying on thin-sliced tissue has limitations in 3D cellular analysis due to structural disruption and tissue loss in the processes of fixation and tissue slicing. Optical sectioning confocal microscopy has been utilized to analyze 3D structure of intact LN tissue without physical tissue slicing. However, light scattering within biological tissues limits the imaging depth only to superficial portion of LN cortex. Recently, optical clearing techniques have shown enhancement of imaging depth in various biological tissues, but their efficacy for LN are remained to be investigated. In this work, we established optical clearing procedure for LN and achieved 3D volumetric visualization of the whole cortex of LN. More than 4 times improvement in imaging depth was confirmed by using LN obtained from H2B-GFP/actin-DsRed double reporter transgenic mouse. With adoptive transfer of GFP expressing B cells and DsRed expressing T cells and fluorescent vascular labeling by anti-CD31 and anti-LYVE-1 antibody conjugates, we successfully visualized major cellularlevel structures such as T-cell zone, B-cell follicle and germinal center. Further, we visualized the GFP expressing metastatic melanoma cell colony, vasculature and lymphatic vessels in the LN cortex.

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High-resolution visualization of mouse cardiac microvasculature using optical histology

Cited by 15 publications

References 43 publications

Optical properties of mouse brain tissue after optical clearing with FocusClear™

Optical properties of mouse brain tissue after optical clearing with FocusClear™

3D imaging in CUBIC-cleared mouse heart tissue: going deeper

Optical clearing based cellular-level 3D visualization of intact lymph node cortex

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