scholarly journals Novel 3D imaging platform tracks cancer progression in vivo

2016 ◽  
Vol 38 (6) ◽  
pp. 12-15
Author(s):  
James McGinty ◽  
Paul French ◽  
Paul Frankel

Optical imaging underpins biomedical research in many respects and recent decades have seen spectacular advances, particularly in fluorescence imaging where genetic engineering approaches to labelling have been combined with new light sources, detectors and data analysis techniques to provide capabilities like super-resolution beyond the diffraction limit, exquisite spectroscopic contrast for molecular readouts and high-speed image capture for in vivo and high-throughput applications. However, the main impact of such advanced instrumentation and data analysis has been to provide unprecedented quantitative 2D and 3D information concerning samples compatible with microscopy where volumes of less than 1 mm3 are typically imaged in a single ‘acquisition’. The ability to view and measure cellular processes and signalling pathways in live cells has been a significant advance for biomedical research and drug discovery. However, for conventional microscope-based assays and experiments, the samples typically comprise thin layers of cells that are not experiencing the same signals that they would in a 3D tissue context and any findings may not directly translate to live organisms. It is desirable to study disease processes in live intact organisms that can provide appropriate physiological complexity. For cancer studies, recent research from our group shows that optical tomography can be used to directly monitor in vivo changes in tumour growth and vascular development in a zebrafish cancer model over time. This technique not only improves the value of the collected data, but if used on a wider scale should result in a reduction in the number of animals used in biomedical research.

2021 ◽  
Vol 9 ◽  
Author(s):  
Shu Zheng ◽  
Yanru Bai ◽  
Zihao Xu ◽  
Pengfei Liu ◽  
Guangjian Ni

Optical coherence tomography (OCT) has become a novel approach to noninvasive imaging in the past three decades, bringing a significant potential to biological research and medical biopsy in situ, particularly in three-dimensional (3D) in vivo conditions. Specifically, OCT systems using broad bandwidth sources, mainly centered at near-infrared-II, allow significantly higher imaging depth, as well as maintain a high-resolution and better signal-to-noise ratio than the traditional microscope, which avoids the scattering blur and thus obtains more details from delicate biological structures not just limited to the surface. Furthermore, OCT systems combined the spectrometer with novel light sources, such as multiplexed superluminescent diodes or ultra-broadband supercontinuum laser sources, to obtain sub-micron resolution imaging with high-speed achieve widespread clinical applications. Besides improving OCT performance, the functional extensions of OCT with other designs and instrumentations, taking polarization state or birefringence into account, have further improved OCT properties and functions. We summarized the conventional principle of OCT systems, including time-domain OCT, Fourier-domain OCT, and several typical OCT extensions, compared their different components and properties, and analyzed factors that affect OCT performance. We also reviewed current applications of OCT in the biomedical field, especially in hearing science, discussed existing limitations and challenges, and looked forward to future development, which may provide a guideline for those with 3D in vivo imaging desires.


2016 ◽  
Vol 2 (5) ◽  
pp. e1600061 ◽  
Author(s):  
Anja Steude ◽  
Emily C. Witts ◽  
Gareth B. Miles ◽  
Malte C. Gather

Optogenetics is a paradigm-changing new method to study and manipulate the behavior of cells with light. Following major advances of the used genetic constructs over the last decade, the light sources required for optogenetic control are now receiving increased attention. We report a novel optogenetic illumination platform based on high-density arrays of microscopic organic light-emitting diodes (OLEDs). Because of the small dimensions of each array element (6 × 9 μm2) and the use of ultrathin device encapsulation, these arrays enable illumination of cells with unprecedented spatiotemporal resolution. We show that adherent eukaryotic cells readily proliferate on these arrays, and we demonstrate specific light-induced control of the ionic current across the membrane of individual live cells expressing different optogenetic constructs. Our work paves the way for the use of OLEDs for cell-specific optogenetic control in cultured neuronal networks and for acute brain slices, or as implants in vivo.


2019 ◽  
Vol 116 (48) ◽  
pp. 24012-24018 ◽  
Author(s):  
Sixian You ◽  
Ronit Barkalifa ◽  
Eric J. Chaney ◽  
Haohua Tu ◽  
Jaena Park ◽  
...  

Despite extensive interest, extracellular vesicle (EV) research remains technically challenging. One of the unexplored gaps in EV research has been the inability to characterize the spatially and functionally heterogeneous populations of EVs based on their metabolic profile. In this paper, we utilize the intrinsic optical metabolic and structural contrast of EVs and demonstrate in vivo/in situ characterization of EVs in a variety of unprocessed (pre)clinical samples. With a pixel-level segmentation mask provided by the deep neural network, individual EVs can be analyzed in terms of their optical signature in the context of their spatial distribution. Quantitative analysis of living tumor-bearing animals and fresh excised human breast tissue revealed abundance of NAD(P)H-rich EVs within the tumor, near the tumor boundary, and around vessel structures. Furthermore, the percentage of NAD(P)H-rich EVs is highly correlated with human breast cancer diagnosis, which emphasizes the important role of metabolic imaging for EV characterization as well as its potential for clinical applications. In addition to the characterization of EV properties, we also demonstrate label-free monitoring of EV dynamics (uptake, release, and movement) in live cells and animals. The in situ metabolic profiling capacity of the proposed method together with the finding of increasing NAD(P)H-rich EV subpopulations in breast cancer have the potential for empowering applications in basic science and enhancing our understanding of the active metabolic roles that EVs play in cancer progression.


Oncotarget ◽  
2016 ◽  
Vol 7 (28) ◽  
pp. 43939-43948 ◽  
Author(s):  
Sunil Kumar ◽  
Nicola Lockwood ◽  
Marie-Christine Ramel ◽  
Teresa Correia ◽  
Matthew Ellis ◽  
...  

2021 ◽  
Vol 118 (37) ◽  
pp. e2015621118
Author(s):  
Yichen Li ◽  
Vasilisa Aksenova ◽  
Mark Tingey ◽  
Jingjie Yu ◽  
Ping Ma ◽  
...  

The in vivo characterization of the exact copy number and the specific function of each composite protein within the nuclear pore complex (NPC) remains both desirable and challenging. Through the implementation of live-cell high-speed super-resolution single-molecule microscopy, we first quantified the native copies of nuclear basket (BSK) proteins (Nup153, Nup50, and Tpr) prior to knocking them down in a highly specific manner via an auxin-inducible degron strategy. Second, we determined the specific roles that BSK proteins play in the nuclear export kinetics of model messenger RNA (mRNA) substrates. Finally, the three-dimensional (3D) nuclear export routes of these mRNA substrates through native NPCs in the absence of specific BSK proteins were obtained and further validated via postlocalization computational simulations. We found that these BSK proteins possess the stoichiometric ratio of 1:1:1 and play distinct roles in the nuclear export of mRNAs within live cells. The absence of Tpr from the NPC predominantly reduces the probability of nuclear mRNAs entering the NPC for export. Complete depletion of Nup153 and Nup50 results in an mRNA nuclear export efficiency decrease of approximately four folds. mRNAs can gain their maximum successful export efficiency as the copy number of Nup153 increased from zero to only half the full complement natively within the NPC. Lastly, the absence of Tpr or Nup153 seems to alter the 3D export routes of mRNAs as they pass through the NPC. However, the removal of Nup50 alone has almost no impact upon mRNA export route and kinetics.


Author(s):  
E. D. Salmon ◽  
J. C. Waters ◽  
C. Waterman-Storer

We have developed a multi-mode digital imaging system which acquires images with a cooled CCD camera (Figure 1). A multiple band pass dichromatic mirror and robotically controlled filter wheels provide wavelength selection for epi-fluorescence. Shutters select illumination either by epi-fluorescence or by transmitted light for phase contrast or DIC. Many of our experiments involve investigations of spindle assembly dynamics and chromosome movements in live cells or unfixed reconstituted preparations in vitro in which photodamage and phototoxicity are major concerns. As a consequence, a major factor in the design was optical efficiency: achieving the highest image quality with the least number of illumination photons. This principle applies to both epi-fluorescence and transmitted light imaging modes. In living cells and extracts, microtubules are visualized using X-rhodamine labeled tubulin. Photoactivation of C2CF-fluorescein labeled tubulin is used to locally mark microtubules in studies of microtubule dynamics and translocation. Chromosomes are labeled with DAPI or Hoechst DNA intercalating dyes.


2012 ◽  
Author(s):  
Dominic Piro ◽  
Kyle A. Brucker ◽  
Thomas T. O'Shea ◽  
Donald Wyatt ◽  
Douglas Dommermuth ◽  
...  

2018 ◽  
Vol 18 (2) ◽  
pp. 156-165 ◽  
Author(s):  
Jiaqiang Wang ◽  
Chien-shan Cheng ◽  
Yan Lu ◽  
Xiaowei Ding ◽  
Minmin Zhu ◽  
...  

Background: Propofol, a widely used intravenous anesthetic agent, is traditionally applied for sedation and general anesthesia. Explanation: Recent attention has been drawn to explore the effect and mechanisms of propofol against cancer progression in vitro and in vivo. Specifically, the proliferation-inhibiting and apoptosis-inducing properties of propofol in cancer have been studied. However, the underlying mechanisms remain unclear. Conclusion: This review focused on the findings within the past ten years and aimed to provide a general overview of propofol's malignance-modulating properties and the potential molecular mechanisms.


2020 ◽  
Vol 13 (3-4) ◽  
pp. 222-228
Author(s):  
И.В. Яминский ◽  
А.И. Ахметова

Разработка высокоэффективных режимов быстродействующего сканирующего зондового микроскопа, в первую очередь атомно-силовой и сканирующей капиллярной микроскопии, представляет особый интерес для успешного проведения биомедицинских исследований: изучения биологических процессов и морфологии биополимеров, определения антибио­тикорезистентности бактерий, адресной доставки биомакромолекул, скринингу лекарств, раннему обнаружению биологических агентов (вирусов и бактерий) и др. The development of highly efficient modes of a high-speed scanning probe microscope, primarily atomic force and scanning capillary microscopy, is of particular interest for successful biomedical research: studying biological processes and the morphology of biopolymers, determining antibiotic resistance of bacteria, targeted delivery of biomacromolecules, drug screening, early detection agents (viruses and bacteria), etc.


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