支持这项研究是由来自美国国家精神分裂症和抑郁症研究联盟和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。海关支持由惠氏CIHR博士后奖学金。作者衷心感谢彼得赛里斯援助，协助数据分析fMRI数据收集和克里斯蒂娜孙明扬。

1。收集和选择回忆，成像任务，和实验协议收集情绪肺泡<ol><li>个人的回忆引起每个参与者在之前进行的fMRI会议接受记者采访时，类似的非情感的AMS（ 例如，11，12）所采用的程序。不像大多数以前的技术，我们的AMQ是专门建造目标评估个人情感的情节和他们recollective的属性。行为研究中常用的方法，利用脑成像研究适应涉及Crovitz和技术希夫曼13日至15日 ，科目记得具体的，亲身经历的事件提示词。一张照片范式也已 ​​用于研究医疗辅助队在fMRI研究16，17更多的控制设置。然而，这些技术通常没有目标检查AMS，recollective素质相匹配的情感成分。 </li><li>我们AMQ包括了115个不同的生活事件（例如，一个家庭成员的出生，亲属死亡）， 这是其他 18，19所使用的组合和扩展列出的口头线索的清单。对于每一个线索，参与者被要求要记住他们的生活的一个独特的情节，发生在一个特定的时间和地点（例如，一个实例时，他/她在一个特定的篮球比赛中发挥），而不是记住一般的或重复的事件（如打高中篮球）。重要的是，必须伴随着回忆亲自参与的回忆，而不是从别人对他们的听力。 </li><li>回忆后，参与者被要求提供了内存的简要说明（见图1），将用于在磁共振成像扫描（参与者天真的预扫描采访的具体目的）作为一种个性化的内存提示。 </li><li>每个内存也是过时和评价六个李克特量表20，21，评估其现象学的属性（见图1）。我们的尺度， 包括情绪价（采用7点量表：-3 =非常消极，0 =中性的，3 =非常积极），情感强度，个人的意义，语境的细节量，visuo知觉的细节量（即，生动性），以及检索的频率 （后者使用7点量表：1 =没有，所有7 =非常） 。 </li></ol><img alt="图1" src="/files/ftp_upload/2396/2396fig1.jpg" /> 图1。 AMQ政府的插图，对于每一个线索，参与者记住并简要描述一个特定的事件，然后日期和6级率。 选择的高度情绪化的肺泡<ol start="5"><li>下一步，每个参与者选择的40个最感性的回忆（20和20负），在AMQ提供的收视率（即评分2或3，分别为-2或-3）的基础上。正面和负面的AMS是那么的年龄和现象属性相匹配，以确保这些基本属性的差异，在回忆中的任何大脑活动的差异不混淆。 </li><li>如果有必要，稍微适应选定的记忆线索的描述要密切的长度和语法的复杂性可能相匹配;还建议选择为实践目的的一些额外的回忆。 </li></ol> fMRI的任务<ol start="7"><li> fMRI的任务是设计，让上午的任务相比，与语义记忆（SM）的控制任务; AM和SM检索试验也有类似的一般结构（见图2A）。我们使用CIGAL（ <a href="http://www.nitrc.org/projects/cigal/">http://www.nitrc.org/projects/cigal/</a>刺激的磁共振扫描仪演示），但也可用于其他刺激演示软件。 </li><li>我的任务是根据扫描前收集的个性化线索。每次试验开始触发上午的回忆，这是由参与者按下一个按钮表示提示。然后，与会者继续回忆事件的细节，直到线索率再次回忆的记忆体（图2B）。 </li><li> SM任务涉及一代的典范，从不同的语义类别（如乐器，体育），这就好比检索涉及搜索在内存和扩展检索时间 22（图2B） 。语义类是线索后，按下一个按钮，尽快为他们开始召回该类别的范例，然后继续回顾直到线索再次内存评级。 </li></ol><img alt="图2" src="/files/ftp_upload/2396/2396fig2a.jpg" /> <img alt="图2" src="/files/ftp_upload/2396/2396fig2b.jpg" /> 图2。结构功能磁共振成像试验。一般结构试验。 B. AM和SM试验的具体结构。 <ol start="10"><li>除了AM和SM试验的基本比较，其他操作也可以参与。例如，可以操纵指挥与会者注意记住的事件的情绪或情感方面（图2B）上午检索的重点。该操作允许调查如何检索的重点可能会影响遇到的AMS和任何相关的变化，在相关的神经关联。 </li><li>其次是每个审判AM或SM评级屏幕呈现在一个平衡的顺序，使用5点李克特量表（见图2b）。 </li><li>实验分成试验运行/块，既允许参与者的时间休息和设备故障的情况下，以避免数据丢失。运行的顺序是参与者之间的平衡。每次运行开始，6秒的固定，使MR信号的稳定。 AM和SM条件审间由一个变量的持续时间间隔（5-9秒，平均= 7sec。​​）分隔的一个随机顺序。 </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。通常情况下，以避免混淆大脑激活偏侧，扫描参与者右手。 在进入扫描室<ol><li>在扫描当天，参加者目前的情感状态是评估 23，控制的心情回忆情感肺泡的影响。结合扫描后评估，这些初步评估，也可以使用屏幕上的情绪变化作为参与研究的结果，以及在功能磁共振成像分析协变量当前状态的影响进行调查的大脑激活。同样，也可以作出评估人格特质（例如，神经质），上午24任务和相关的神经关联探讨其可能出现的偏差。 </li><li>扫描之前，参加者在扫描程序的详细通知，并给予行为任务的具体说明。与会者还完成一个简短的练习，熟悉自己的任务。 </li></ol>进入扫描室<ol start="3"><li>与会者指示仰卧躺在扫描床上，并提供额外的头部缓冲，以确保扫描过程中的舒适性，并尽量减少运动。为了进一步减少头部运动，非粘合面的长度的磁带可能被包左右的受试者的前额轻轻。科目耳保护以及隔离耳机与沟通，在MRI扫描实验者。 </li><li>主题的右手定位舒适的响应中，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。 </li><li>开始收集数据之前，它是确保科目，可以看到屏幕投影清楚地刺激呈现和响应按钮正常工作的关键。 </li></ol> 3。数据记录和处理扫描参数我们收集使用1.5特斯拉西门子奏鸣曲扫描器的核磁共振MRI检查记录数据。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;片数= 112;体素大小= 1x1x1毫米），和功能图像系列28个功能片，收购轴向使用echoplanar序列（TR = 2000毫秒，TE = 40毫秒，实地查看FOV = 256 × 256毫米;体素大小为4x4x4毫米），从而使覆盖全脑。 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> ）相结合，与内部基于Matlab工具。前处理的典型步骤：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） 。个人和组级的统计分析可能涉及的大脑活动进行比较，根据内存的类型（上午与SM），情绪价（正与负），和检索的重点（情感与非情感的内容）。 4。代表性的成果<img alt="图3" src="/files/ftp_upload/2396/2396fig3.jpg" /> 图3。上午检索神经关联。验证本议定书，AMS的检索取得了上午检索网络4，25，包括河马活动增加campal领域（一），在一般的记忆检索，内侧前额叶皮质（二）个人参与，cuneus /楔前叶区域和parieto枕交界处（C，E，分别），涉及与处理visuo空间表述相关额颞交界处（D），涉及情感上午检索;后者类似的效果还发现杏仁核（未显示）。 “激活地图”叠加高分辨率脑冠状（左图）显示的图像和矢状（中间和右边的面板）的意见;彩条显示激活地图的 t值梯度（P &lt;0.005，连续10体素26），反映大脑活动的时间锁定到内存线索onsets。线图说明的fMRI信号的时间制课程（％信号变化），每次试验的类型和TR（1 TR = 2秒）。 L =左，R =右。

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没有利益冲突的声明。

这里介绍实验设计允许的记忆情感的自传体记忆的神经相关性的调查。这种设计有可能推进我们的大脑如何产生情感偏见（正或负），在记忆的个人记忆的知识，以及如何将这些偏差可能是由检索的重点调制（情绪或情感方面）。该协议有额外的好处，它也可用于临床的人群（如抑郁症和创伤后应激障碍患者），这使得调查上午检索的负面情感偏见的改变（例如，负面的经验反刍创伤性事件和不?...

brain imaging investigation of the neural correlates of emotional autobiographical recollection

情感自传式记忆体（AMS）的回忆，重要的是健康的认知和情感的运作1 -记住积极AMS是增加个人的福祉和自尊2，而可能会导致记忆和负面的 AM反刍3情感性精神障碍。虽然已取得重大进展，在了解底层AM一般的检索（4，5审查），少的是情感的主观上的AMS和相关的神经关联重新体验的影响已知的脑机制。这部分是由于这样的事实，不像实验室为基础的微观活动（6，7-9审查）内存情感效应的调查，很多时候，我的研究没有明确的重点在情感方面记住个人事件（见10）。在这里，我们提出了一个协议，允许调查回味的情感队使用功能性磁共振成像（fMRI）的神经关联。这些记忆的线索收集，扫描前问卷（AMQ）的自传体记忆的手段，因此，基于现象学的属性（即强度，生动性，个人的意义）选择适当的情感肺泡。该协议可用于健康和临床人群的一致好评。

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Brain Imaging Investigation of the Neural Correlates of Emotional Autobiographical Recollection

我们提出了一个协议，允许调查回味的情感自传体记忆体，使用功能磁共振成像的神经关联。该协议可用于健康和临床参与者。

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-neural-correlates-emotional

Research

JoVE Journal

Neuroscience

9.7K 次观看.  University of Alberta, Edmonton. 我们提出了一个协议，允许调查回味的情感自传体记忆体，使用功能磁共振成像的神经关联。该协议可用于健康和临床参与者。

视频: Brain Imaging Investigation of the Neural Correlates of Emotional Autobiographical Recollection

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 Emotion Regulation

The ability to control/regulate emotions is an important coping mechanism in the face of emotionally stressful situations. Although significant progress has been made in understanding conscious/deliberate emotion regulation (ER), less is known about non-conscious/automatic ER and the associated neural correlates. This is in part due to the problems inherent in the unitary concepts of automatic and conscious processing1. Here, we present a protocol that allows investigation of the neural correlates of both deliberate and automatic ER using functional magnetic resonance imaging (fMRI). This protocol allows new avenues of inquiry into various aspects of ER. For instance, the experimental design allows manipulation of the goal to regulate emotion (conscious vs. non-conscious), as well as the intensity of the emotional challenge (high vs. low). Moreover, it allows investigation of both immediate (emotion perception) and long-term effects (emotional memory) of ER strategies on emotion processing. Therefore, this protocol may contribute to better understanding of the neural mechanisms of emotion regulation in healthy behaviour, and to gaining insight into possible causes of deficits in depression and anxiety disorders in which emotion dysregulation is often among the core debilitating features.

We present a protocol that allows investigation of the neural correlates of deliberate and automatic emotion regulation, using functional magnetic resonance imaging. This protocol can be used in healthy participants, both young and older, as well as in clinical patients.

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 ...

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 ...

Detection of Microregional Hypoxia in Mouse Cerebral Cortex by Two-photon Imaging of Endogenous NADH Fluorescence

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. Here we present a visualized experimental and methodological protocol to directly visualize microregional tissue hypoxia and to infer perivascular oxygen gradients in the mouse cortex. It is based on the non-linear relationship between nicotinamide adenine dinucleotide (NADH) endogenous fluorescence intensity and oxygen partial pressure in the tissue, where observed tissue NADH fluorescence abruptly increases at tissue oxygen levels below 10 mmHg1. We use two-photon excitation at 740 nm which allows for concurrent excitation of intrinsic NADH tissue fluorescence and blood plasma contrasted with Texas-Red dextran. The advantages of this method over existing approaches include the following: it takes advantage of an intrinsic tissue signal and can be performed using standard two-photon in vivo imaging equipment; it permits continuous monitoring in the whole field of view with a depth resolution of ~50 &mu;m. We demonstrate that brain tissue areas furthest from cerebral blood vessels correspond to vulnerable watershed areas which are the first to become functionally hypoxic following a decline in vascular oxygen supply. This method allows one to image microregional cortical oxygenation and is therefore useful for examining the role of inadequate or restricted tissue oxygen supply in neurovascular diseases and stroke.

Here we describe a method to directly visualize microregional tissue hypoxia in the mouse cortex in vivo. It is based on concurrent two-photon imaging of nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation. This method is useful for high resolution analysis of tissue oxygen supply.

检测内源NADH的荧光双光子成像的小鼠大脑皮层Microregional缺氧

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. ...

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 ...

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 ...