支持这项研究是由来自全国联盟为精神分裂症和抑郁症的研究，并从加拿大精神病学研究基金会（FD）的俄共奖的青年研究者奖。作者衷心感谢援助，协助数据分析fMRI数据收集和克里斯蒂娜孙明扬特丽莎恰克拉巴提和彼得赛里斯。

1。任务设计，刺激，实验协议总体协议涉及的图片（ 直接影响）的情感收视率的评估，并为内存 这些图片（ 长期影响），作为诱导目标，以调节情绪反应的结果。 或明或暗地进行规范的目标是诱发，和眼前的和长远的影响评估相对收视率和内存在基线块/ ER的操作（前ER感应基线）之前的运行呈现的图片，见图1整体实验设计图。下面我们提供了所有这些方面的细节。 <img alt="图1" src="/files/ftp_upload/2430/2430fig1.jpg" /> 图1。图诱导内质网的目标是在每个参与者都自觉地/明确和non-consciously/implicitly，但归纳的顺序是不同的，和参与者之间平衡的协议-即那些在第一部分中明确的ER完成的隐在第二部分中，反之亦然;操纵每个操作之前，是由自身的基线运行。评价和记忆任务的详细描述见下文。 情绪调节操作<ol><li> 明确ER的操纵 。在明确的ER状况，情感评级任务指令之前压抑的情感体验和表达情绪反应。以前的证据表明，明确ER的战略能够可靠地影响情绪响应2。 </li><li> 隐ER的操纵 。对于这一点，适应炒句子任务（SST）的3是用于非自觉诱发调节情绪的目标。在此任务中，参与者有20 4个字的语法正确的句子构造嵌入式传达的情绪控制的想法（例如，“限制”，“稳定”，“盖”）的话从五字jumbles，从而总理与会者压抑自己的情绪反应4。从以往的研究4，采取了情绪控制的话，要求本科的学生选择“列出的20个字来你的头脑，当你想到”情绪控制“的概念。“ </li><li>值得注意的是，要维护的结构相同的任务，在双方的ER条件下，SST也是明确的ER条件，但在这种情况下，参与者与20只的目标，控制情绪的中性用语的句子。 </li></ol>操纵的ER直接的影响进行评估：情绪评价任务<ol start="4"><li> 刺激的选择。刺激包括从国际情感图片系统（IAPS）5基于情绪价和觉醒的规范性分数，选定的情感和中性图片。注意平衡的图片在所有研究运行的情感内容，从而避免由于可能存在的差异，在这些基本属性混杂影响。 </li><li> 定时任务 。每张照片上的4秒钟的画面，然后被删除，以尽量减少眼球运动相关的长时间扫描图像 6混杂的影响;一个固定跨如下每12秒的画面。参加者的任务是观看图片，并为自己的主观情感体验，触发8点规模的图片（1 =中性，8 =极其不利）。为了避免情绪感应，图片是伪随机的，所以，相同价不超过三个图片连续呈现。 </li></ol>评估的ER操纵的长期影响：记忆提取任务<ol start="6"><li>一个星期后，与会者认为旧的数目相等（见扫描仪） 和新图片（以前从未见过的），在黑色和白色，以防止的天花板性能。 新/旧的决定基于他们是否记得看到图片而在扫描仪或，与会者还率在3点规模的信心，他们的反应（1 =低2 =中3 =高可信度） 。 </li></ol> 2。准备扫描的主题所有的参与者提供书面知情同意书之前，运行实验的协议，这是由伦理委员会的批准。他们还警告说，许多图片描绘了令人痛心的事件，如暴力或创伤的行为，，并提供印有代表性的图片的例子。 扫描前<ol><li>扫描一天，参与者的当前状态“的心态，是评估 7，以控制为T他情绪影响的情感体验和控制情绪的能力。结合扫描后评估，初步评估也可用于情绪变化屏幕作为参与研究的结果。这些索引习惯性的情绪调节策略8参与评估的个性特征，也可以进行ER操作和相关的神经关联的成功，以探讨其可能产生的影响。 </li><li>扫描前，与会者被告知详细的扫描程序，行为的工作给予了具体的指示。为了避免不适和创建任务的熟悉程度增加，参加者也略实践情感评价任务和SST的运行。 </li></ol>在扫描室<ol start="3"><li>与会者指示躺在扫描床上仰卧，头额外的缓冲，以确保在扫描过程中的舒适性，并尽量减少运动。非粘合面的长度磁带可能的参与者的额头上轻轻裹住头部运动，以进一步减少。与会者耳保护以及隔离耳机与沟通，在MRI扫描实验者。 </li><li>受试者的右手定位舒适响应盒上，从而使左手被用于支持或其他测量（例如，皮肤电导反应）。是放在附近的一个紧急停止按钮，使这个问题可能表示任何迫切需要停止扫描仪。在开始收集数据之前，它是至关重要的，以确保主体清楚地看到屏幕投影，刺激呈现和响应按钮的正常工作。 </li><li>这项研究分为试验运行/块，让参与者的休息时间，并在设备故障的情况下，以避免大量的数据损失。运行的顺序是平衡参与者之间，每次运行开始6秒的固定，允许​​MR信号稳定。 </li></ol> 3。数据记录和处理扫描参数我们收集了从24岁的年轻健康受试者的核磁共振MRI检查记录数据，使用一个1.5特斯拉的西门子奏鸣曲扫描器。我们的解剖图像的三维MPRAGE解剖系列（重复时间（TR）= 1600毫秒，回波时间（TE）= 3.82毫秒;片= 112;体素大小= 1x1x1mm）。收购轴向使用echoplanar序列（TR = 2000毫秒，TE = 40 ms的角度视场= 256x256mm领域），从而使覆盖全脑功能图像系列28片（体素大小为4x4x4毫米）功能， 。 数据分析我们用统计参数图（SPM： <a href="http://www.fil.ion.ucl.ac.uk/spm/">http://www.fil.ion.ucl.ac.uk/spm/</a> ）与内部基于MATLAB的工具组合。前处理涉及的典型步骤：质量保证，TR对齐，运动校正，共同登记，规范化，和平滑的（8毫米3内核）9。据急诊室操纵（ER与基线运行;见图2），情绪价（负与中性），觉醒（低 - 高），和内存性能，个人和组级的统计分析可能涉及的大脑活动进行比较（记得与遗忘）。行为数据（如图片评分）和/或分数索引个性措施（例如，索引的ER战略的习惯性接触的人格特质）的脑成像数据的相关性也可以进行，大脑活动与个体差异，探讨如何合作不同行为和人格。 4。代表性的成果： <img alt="图2" src="/files/ftp_upload/2430/2430fig2.jpg" /> 图2。减少杏仁核活动，并增加了前额叶皮层的活动ER的操纵。诱导的ER目标是与情绪加工相关的脑区，包括杏仁核（一）相关的活动减少，并增加与认知控制和情绪调节相关的大脑区域的活动。 10，包括腹外侧前额叶皮层（PFC）（b）项，背外侧PFC（D），与内侧PFC（C）11,12。栏说明vlPFC活动图，明确的和隐含的ER响应类似的模式，这表明，这些变化反映了显性和隐性的ER联合贡献。叠加“激活地图”上的高分辨率图像显示大脑冠状意见;彩条显示激活地图的t值的梯度，根据统计组（N = 24），反映中显示下降的体素的大脑活动（左侧面板）或增加（右图）活动的ER操纵的结果。应该指出，在PFC激活斑点可能会覆盖更多地区，如下：vlPFC /岛（b）和背侧前扣带皮层/运动前皮层（C）。条形图说明情绪（EMO）和中性（NEU）的照片之前（即BaseRuns）和诱导后的ER目标（即％信号的变化，结合ER =隐和外显的急诊室平均起来，明确急诊室，隐ER），显示后ER差异的前体素的提取。 L =左，R =右。 <img alt="表1" src="/files/ftp_upload/2430/2430table1.jpg" /> 表1。以下ER操纵的情感评级下降 。类似的功能磁共振成像的结果，有减少雌激素受体 ​​诱导的情感评级，这起跌幅导致下面的显性和隐性的ER。重要的是，情感收视率没有发生明显变化，在对照组，执行相同的任务，以下类似的延误，但在ER目标归纳的情况下。支持这些想法，2（操作：基线与ER感应）× 2（价：情绪与中性）从实验组*的数据的重复测量方差分析，取得了显着操纵的主要效果（F [1，19] （N = 9）= 11.21，P &lt;0.002）和显着操纵x价的交互作用（F [1，19] = 3.02，P &lt;0.05），也有类似的2路从对照组的数据方差没有屈服主效应显着的延迟（基线与延迟运行）或明显的延迟x价互动。这些结果表明，在实验组的收视率下降是由于诱导目标，控制情绪反复接触后情绪刺激，而不是由于习惯。 *由于数据流失的常见原因（例如，技术故障），因为，在实验组的行为分析的基础上，从20名参加者的数据。 两者合计，这些行为和脑成像结果验证本实验的设计，它可以用来比较显性和隐性的目标归纳，以调节情绪。应该指出，本报告强调方面所造成的显性和隐性的ER操作的联合贡献，但这并不排除这种可能性，其影响可能会在行为和/或脑水平的可分离。

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没有利益冲突的声明。

我们描述了一个协议，它的目标包括显性和隐性的操纵来调节情绪，并允许调查相关的神经关联。这种设计有可能推进我们的大脑是如何调节情绪，适合比较明确的ER隐含的急诊室，这是无意的，可能发生不参与者的见解和认识 13,14知识。因此，该协议可为惯性相反的情感偏见，如在健康老龄化（ 阳性偏置15）和抑郁（ 消极偏见16）观察到的，这是与ER的机制...

brain imaging investigation of the neural correlates of emotion regulation

控制/调节情绪的能力是一个重要的的应对机制，在面对情绪困扰的情况下。虽然已取得重大进展，理解意识/故意情绪调节（ER）的少了解non-conscious/automatic ER和相关的神经关联。这是因为在自动和自觉处理1的单一概念所固有的问题的一部分。在这里，我们提出了一个协议，允许调查故意和自动急诊室使用功能磁共振成像（fMRI）的神经关联。此协议允许各方面的ER调查的新途径。例如，实验设计，使操纵的目标，调节情感（自觉与不自觉的），以及情感的挑战（高与低）的强度。此外，允许双方立即情绪知觉和长期影响情绪记忆的情绪加工的ER战略展开调查。因此，这个协议可能有助于更好地理解情绪调节健康的行为的神经机制，并获得洞察到，抑郁症和焦虑症的赤字情感DYS监管往往是衰弱的功能之间的核心的可能原因。

Watch this Scientific Journal Video about Brain Imaging Investigation of the Neural Correlates of Emotion Regulation at JoVE.com

Brain Imaging Investigation of the Neural Correlates of Emotion Regulation

我们提出了一个协议，允许调查故意和自动情绪调节，使用功能磁共振成像，神经关联。该协议可用于在健康的参与者，年轻人和老年人，以及在临床病人。

情绪调节的神经关联的脑成像研究

The ability to control/regulate emotions is an important coping mechanism in the face of emotionally stressful situations. Although significant progress ...

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12.3K 次观看.  University of Illinois, Urbana-Champaign. 我们提出了一个协议，允许调查故意和自动情绪调节，使用功能磁共振成像，神经关联。该协议可用于在健康的参与者，年轻人和老年人，以及在临床病人。

视频: Brain Imaging Investigation of the Neural Correlates of Emotion Regulation

Brain Imaging Investigation of the Neural Correlates of Observing Virtual Social Interactions

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language affect our decisions in personal and professional life alike 1. These "friend or foe" judgements are often based on first impressions, which in turn may affect our decisions to "approach or avoid". Previous studies investigating the neural correlates of social cognition tended to use static facial stimuli 2. Here, we illustrate an experimental design in which whole-body animated characters were used in conjunction with functional magnetic resonance imaging (fMRI) recordings. Fifteen participants were presented with short movie-clips of guest-host interactions in a business setting, while fMRI data were recorded; at the end of each movie, participants also provided ratings of the host behaviour. This design mimics more closely real-life situations, and hence may contribute to better understanding of the neural mechanisms of social interactions in healthy behaviour, and to gaining insight into possible causes of deficits in social behaviour in such clinical conditions as social anxiety and autism 3.

This article demonstrates an experimental design in which whole-body animated characters are used in conjunction with functional magnetic resonance imaging (fMRI) to investigate the neural correlates of observing virtual social interactions.

观察虚拟社会交往中的神经关联的脑成像研究

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language ...

科研

神经科学

Brain Imaging Investigation of the Neural Correlates of Emotional Autobiographical Recollection

Recollection of emotional autobiographical memories (AMs) is important to healthy cognitive and affective functioning 1 - remembering positive AMs is associated with increased personal well-being and self-esteem 2, whereas remembering and ruminating on negative AMs may lead to affective disorders 3. Although significant progress has been made in understanding the brain mechanisms underlying AM retrieval in general (reviewed in 4, 5), less is known about the effect of emotion on the subjective re-experience of AMs and the associated neural correlates. This is in part due to the fact that, unlike the investigations of the emotion effect on memory for laboratory-based microevents (reviewed in 6, 7-9), often times AM studies do not have a clear focus on the emotional aspects of remembering personal events (but see 10). Here, we present a protocol that allows investigation of the neural correlates of recollecting emotional AMs using functional magnetic resonance imaging (fMRI). Cues for these memories are collected prior to scanning by means of an autobiographical memory questionnaire (AMQ), therefore allowing for proper selection of emotional AMs based on their phenomenological properties (i.e., intensity, vividness, personal significance). This protocol can be used in healthy and clinical populations alike.

We present a protocol that allows investigation of the neural correlates of recollecting emotional autobiographical memories, using functional magnetic resonance imaging. This protocol can be used with both healthy and clinical participants.

情感自传回忆的神经关联的脑成像研究

Recollection of emotional autobiographical memories (AMs) is important to healthy cognitive and affective functioning 1 - remembering positive AMs is ...

Brain Imaging Investigation of the Memory-Enhancing Effect of Emotion

Emotional events tend to be better remembered than non-emotional events1,2. One goal of cognitive and affective neuroscientists is 
to understand the neural mechanisms underlying this enhancing effect of emotion on memory. A method that has proven particularly influential in the 
investigation of the memory-enhancing effect of emotion is the so-called subsequent memory paradigm (SMP). This method was originally used to investigate the 
neural correlates of non-emotional memories3, and more recently we and others also applied it successfully to studies of emotional memory (reviewed in4, 5-7). 
Here, we describe a protocol that allows investigation of the neural correlates of the memory-enhancing effect of emotion using the SMP in conjunction with 
event-related functional magnetic resonance imaging (fMRI). An important feature of the SMP is that it allows separation of brain activity specifically 
associated with memory from more general activity associated with perception. Moreover, in the context of investigating the impact of emotional stimuli, 
SMP allows identification of brain regions whose activity is susceptible to emotional modulation of both general/perceptual and memory-specific processing. 
This protocol can be used in healthy subjects8-15, as well as in clinical patients where there are alterations in the neural correlates of emotion perception 
and biases in remembering emotional events, such as those suffering from depression and post-traumatic stress disorder (PTSD)16, 17.

We present a protocol that uses functional magnetic resonance imaging to investigate the neural correlates of the memory-enhancing effect of emotion. This protocol allows identification of brain activity specifically linked to memory-related processing, contrary to more general perceptual processing, and can be used with healthy and clinical populations.

脑成像的情感记忆的促进作用调查

Emotional events tend to be better remembered than non-emotional events1,2. One goal of cognitive and affective neuroscientists is 
to understand the ...

Brain Imaging Investigation of the Impairing Effect of Emotion on Cognition

Emotions can impact cognition by exerting both enhancing (e.g., better memory for emotional events) and impairing (e.g., increased emotional distractibility) effects (reviewed in 1). Complementing our recent protocol 2 describing a method that allows investigation of the neural correlates of the memory-enhancing effect of emotion (see also 1, 3-5), here we present a protocol that allows investigation of the neural correlates of the detrimental impact of emotion on cognition. The main feature of this method is that it allows identification of reciprocal modulations between activity in a ventral neural system, involved in 'hot' emotion processing (HotEmo system), and a dorsal system, involved in higher-level 'cold' cognitive/executive processing (ColdEx system), which are linked to cognitive performance and to individual variations in behavior (reviewed in 1). Since its initial introduction 6, this design has proven particularly versatile and influential in the elucidation of various aspects concerning the neural correlates of the detrimental impact of emotional distraction on cognition, with a focus on working memory (WM), and of coping with such distraction 7,11, in both healthy 8-11 and clinical participants 12-14.

We present a protocol that allows investigation of the neural mechanisms mediating the detrimental impact of emotion on cognition, using functional magnetic resonance imaging. This protocol can be used with both healthy and clinical participants.

脑成像的情感对认知损害的影响调查

Emotions can impact cognition by exerting both enhancing (e.g., better memory for emotional events) and impairing (e.g., increased emotional ...

Extinction Training During the Reconsolidation Window Prevents Recovery of Fear

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents the return of fear. Pavlovian fear-conditioning paradigms are commonly employed to create a controlled, novel fear association in the laboratory. After pairing an innocuous stimulus (conditioned stimulus, CS) with an aversive outcome (unconditioned stimulus, US) we can elicit a fear response (conditioned response, or CR) by presenting just the stimulus alone1,2 . Once fear is acquired, it can be diminished using extinction training, whereby the conditioned stimulus is repeatedly presented without the aversive outcome until fear is no longer expressed3. This inhibitory learning creates a new, safe representation for the CS, which competes for expression with the original fear memory4. Although extinction is effective at inhibiting fear, it is not permanent. Fear can spontaneously recover with the passage of time. Exposure to stress or returning to the context of initial learning can also cause fear to resurface3,4.
Our protocol addresses the transient nature of extinction by targeting the reconsolidation window to modify emotional memory in a more permanent manner. Ample evidence suggests that reactivating a consolidated memory returns it to a labile state, during which the memory is again susceptible to interference5-9. This window of opportunity appears to open shortly after reactivation and close approximately 6hrs later5,11,16, although this may vary depending on the strength and age of the memory15. By allowing new information to incorporate into the original memory trace, this memory may be updated as it reconsolidates10,11. Studies involving non-human animals have successfully blocked the expression of fear memory by introducing pharmacological manipulations within the reconsolidation window, however, most agents used are either toxic to humans or show equivocal effects when used in human studies12-14. Our protocol addresses these challenges by offering an effective, yet non-invasive, behavioral manipulation that is safe for humans.
By prompting fear memory retrieval prior to extinction, we essentially trigger the reconsolidation process, allowing new safety information (i.e., extinction) to be incorporated while the fear memory is still susceptible to interference. A recent study employing this behavioral manipulation in rats has successfully blocked fear memory using these temporal parameters11. Additional studies in humans have demonstrated that introducing new information after the retrieval of previously consolidated motor16, episodic17, or declarative18 memories leads to interference with the original memory trace14. We outline below a novel protocol used to block fear recovery in humans.

Conditioned fear can be diminished through an inhibitory process called extinction, but can resurface under conditions such as the passage of time or exposure to stress. Our protocol presents a novel way of preventing fear recovery by introducing extinction during the reconsolidation window (the re-storage phase of a reactivated memory).

灭绝培训在彭兰霞窗口无法还原的恐惧

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents ...

High-resolution Live Imaging of Cell Behavior in the Developing Neuroepithelium

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central nervous system (CNS). Although much is known about the molecular mechanisms that pattern the spinal cord and elicit neuronal differentiation1, 2, we lack a deep understanding of these early events at the level of cell behavior. It is thus critical to study the behavior of neural progenitors in real time as they undergo neurogenesis.
In the past, real-time imaging of early embryonic tissue has been limited by cell/tissue viability in culture as well as the phototoxic effects of fluorescent imaging. Here we present a novel assay for imaging such tissue for long periods of time, utilizing a novel ex vivo slice culture protocol and wide-field fluorescence microscopy (Fig. 1). This approach achieves long-term time-lapse monitoring of chick embryonic spinal cord progenitor cells with high spatial and temporal resolution.
This assay may be modified to image a range of embryonic tissues3, 4 In addition to the observation of cellular and sub-cellular behaviors, the development of novel and highly sensitive reporters for gene activity (for example, Notch signaling5) makes this assay a powerful tool with which to understand how signaling regulates cell behavior during embryonic development.

Imaging embryonic tissue in real-time is challenging over long periods of time. Here we present an assay for monitoring cellular and sub-cellular changes in chick spinal cord for long periods with high spatial and temporal resolution. This technique can be adapted for other regions of the nervous system and developing embryo.

高分辨率实时成像发展中的神经上皮细胞的行为

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central ...

Co-analysis of Brain Structure and Function using fMRI and Diffusion-weighted Imaging

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when studying the brain, as the functional role that a brain area fulfills may be largely defined by its connections to other brain areas. In this report, we describe a novel, non-invasive approach for relating brain structure and function using magnetic resonance imaging (MRI). This approach, a combination of structural imaging of long-range fiber connections and functional imaging data, is illustrated in two distinct cognitive domains, visual attention and face perception. Structural imaging is performed with diffusion-weighted imaging (DWI) and fiber tractography, which track the diffusion of water molecules along white-matter fiber tracts in the brain (Figure 1). By visualizing these fiber tracts, we are able to investigate the long-range connective architecture of the brain. The results compare favorably with one of the most widely-used techniques in DWI, diffusion tensor imaging (DTI). DTI is unable to resolve complex configurations of fiber tracts, limiting its utility for constructing detailed, anatomically-informed models of brain function. In contrast, our analyses reproduce known neuroanatomy with precision and accuracy. This advantage is partly due to data acquisition procedures: while many DTI protocols measure diffusion in a small number of directions (e.g., 6 or 12), we employ a diffusion spectrum imaging (DSI)1, 2 protocol which assesses diffusion in 257 directions and at a range of magnetic gradient strengths. Moreover, DSI data allow us to use more sophisticated methods for reconstructing acquired data. In two experiments (visual attention and face perception), tractography reveals that co-active areas of the human brain are anatomically connected, supporting extant hypotheses that they form functional networks. DWI allows us to create a "circuit diagram" and reproduce it on an individual-subject basis, for the purpose of monitoring task-relevant brain activity in networks of interest.

We describe a novel approach for simultaneous analysis of brain function and structure using magnetic resonance imaging (MRI). We assess brain structure with high-resolution diffusion-weighted imaging and white-matter fiber tractography. Unlike standard structural MRI, these techniques allow us to directly relate anatomical connectivity to functional properties of brain networks.

使用功能磁共振成像和弥散加权成像脑结构与功能分析

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when ...

A Computer-assisted Multi-electrode Patch-clamp System

The patch-clamp technique is today the most well-established method for recording electrical activity from individual neurons or their subcellular compartments. Nevertheless, achieving stable recordings, even from individual cells, remains a time-consuming procedure of considerable complexity. Automation of many steps in conjunction with efficient information display can greatly assist experimentalists in performing a larger number of recordings with greater reliability and in less time. In order to achieve large-scale recordings we concluded the most efficient approach is not to fully automatize the process but to simplify the experimental steps and reduce the chances of human error while efficiently incorporating the experimenter's experience and visual feedback. With these goals in mind we developed a computer-assisted system which centralizes all the controls necessary for a multi-electrode patch-clamp experiment in a single interface, a commercially available wireless gamepad, while displaying experiment related information and guidance cues on the computer screen. Here we describe the different components of the system which allowed us to reduce the time required for achieving the recording configuration and substantially increase the chances of successfully recording large numbers of neurons simultaneously.

Multi-electrode patch-clamp recordings constitute a complex task. Here we show how, by automating of many of the experimental steps, it is possible to accelerate the process leading to qualitative improvement in performance and number of recordings.

计算机辅助多电极膜片钳系统

The patch-clamp technique is today the most  well-established method for recording electrical activity from individual  neurons or their subcellular ...

Analysis of Brain Mitochondria Using Serial Block-Face Scanning Electron Microscopy

Human brain is a high energy consuming organ that mainly relies on glucose as a fuel source. Glucose is catabolized by brain mitochondria via glycolysis, tri-carboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) pathways to produce cellular energy in the form of adenosine triphosphate (ATP). Impairment of mitochondrial ATP production causes mitochondrial disorders, which present clinically with prominent neurological and myopathic symptoms. Mitochondrial defects are also present in neurodevelopmental disorders (e.g. autism spectrum disorder) and neurodegenerative disorders (e.g. amyotrophic lateral sclerosis, Alzheimer&#39;s and Parkinson&#39;s diseases). Thus, there is an increased interest in the field for performing 3D analysis of mitochondrial morphology, structure and distribution under both healthy and disease states. The brain mitochondrial morphology is extremely diverse, with some mitochondria especially those in the synaptic region being in the range of &#60;200 nm diameter, which is below the resolution limit of traditional light microscopy. Expressing a mitochondrially-targeted green fluorescent protein (GFP) in the brain significantly enhances the organellar detection by confocal microscopy. However, it does not overcome the constraints on the sensitivity of detection of relatively small sized mitochondria without oversaturating the images of large sized mitochondria. While serial transmission electron microscopy has been successfully used to characterize mitochondria at the neuronal synapse, this technique is extremely time-consuming especially when comparing multiple samples. The serial block-face scanning electron microscopy (SBFSEM) technique involves an automated process of sectioning, imaging blocks of tissue and data acquisition. Here, we provide a protocol to perform SBFSEM of a defined region from rodent brain to rapidly reconstruct and visualize mitochondrial morphology. This technique could also be used to provide accurate information on mitochondrial number, volume, size and distribution in a defined brain region. Since the obtained image resolution is high (typically under 10 nm) any gross mitochondrial morphological defects may also be detected.

Mitochondrial visualization and analysis from mammalian brain tissue is a challenging task. Here, we describe how three dimensional (3D) reconstruction analysis from the serial block-face scanning electron microscopy (SBFSEM) can be used to gain insights on the morphological and volumetric analysis of this critical energy generating organelle.

使用串行块面部扫描电子显微镜脑线粒体分析

Human brain is a high energy consuming organ that mainly relies on glucose as a fuel source. Glucose is catabolized by brain mitochondria via glycolysis, ...

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in adult mammalian animals remains elusive. In this study, we describe an experimental procedure for measuring calcium dynamics through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy. This method enables recordings and quantifications of neural activity in bilateral mouse brain regions one at a time in the same experiment for a prolonged period in vivo. Key aspects of this method, which can be completed within an hour, include minimally invasive surgery procedures for creating dual optical windows, and the use of 2P imaging. Although we only demonstrate the technique in the S1 area, the method can be applied to other regions of the living brain facilitating the elucidation of structural and functional complexities of brain neural networks.

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo.

胸腺1-gmcmp6 转基因小鼠在原代体血源皮质中的成像神经活性

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...