저자는 자신의 지속적인 기술 지원 Ikiel Ptito 감사하고 싶습니다.

1 부 : 조직의 사전 처리 <ol><li> 조직은 잘 paraformaldehyde, 글루 타 알데히드, 또는 포르말린과 perfused해야합니다. 이것은 일반적으로 다른 장기를 수확하는 데 사용되는 표준 transcardial 재관류를 통해 달성될 수있다. 현재 연구에서 주제가 깊이 나트륨 pentobarbital (25 MG / kg, IV)의 과다로 euthanized 케타민을 하이드로 클로라이드 (10 MG / kg, IM)과 진정했고 완전히 exsanguinated 때까지 0.1 M PBS로 transcardially perfused. 이것은 5 분 (~ 1리터)을 PBS에 4 % paraformaldehyde 솔루션옵니다. </li><li> 두뇌는 stereotaxically, 차단 두개골에서 제거, 무게, 볼륨 2 결정되어야합니다. 조직은 다음 인산염 버퍼의 30 % 자당의 최종 농도로 등급 자당 솔루션에 cryoprotected해야합니다. 조직 단위가 다음 -65 ° C에서 이소펜탄에 냉동과 -80에 보관해야 ° C가 슬라이스 준비가 될 때까지. </li></ol> 2 부 : 체계적인 샘플링 <ol><li> 체계적인 샘플링 이전 sectioning에 대한 적절한 계획에 의존합니다. 주동이의 - 꼬리와 지느러미 - 복부 extents 모두에 대한 관심의 전체 영역이 잘 정의되어야한다. 사용 종족의 stereotaxic 아틀라스는 관심의 영역을 정의하고 관심 영역의 길이를 결정하기 위해 도움이됩니다. 일단 관심 영역의 길이는 샘플링 간격을 설정하는 데 필요한 결정됩니다. 예를 들어, stereotaxic 아틀라스에서 당신은 코로나 비행기에 sectioning 때 관심 영역의 전체 rostro - 꼬리 정도 길이가 약 3mm 것을 확인하십시오. 그런 다음, 50μm에서 섹션에 그라 이오 스탯을 설정하려면이 속도에서 관심의 영역의 주동이의 - 꼬리 범위에 걸쳐 60 섹션을 결정합니다. 일반적인 편견 stereological 연구 60-10 체계적으로 샘플 섹션은 신뢰할 수있는 결과를 얻기 위해 필요합니다. 이 예제에서 10 섹션은 동일 육분의 일 섹션의 샘플링 간격의 rostro - 꼬리 어느 정도 결과를 통해 간격. </li></ol> 파트 3 : 두뇌 은행의 창조 <ol><li> 섹션 샘플링 주파수가 결정되면, 다음 단계는 얼룩이 즉시 수행할지 결정하는 것입니다. 예를 들어, cresyl 제비꽃과 얼룩있다면 당신은 슬라이드에서 육분의 일 섹션을 사로잡을 것입니다. 이 시리즈 1 것입니다. 당신이 원하는 것을 아마도 몇 가지 잠재적인 항체가 알고있다면 실행 그렇다고 기본 얼룩을 제공하거나 immunohistochemistry을 수행할 수있는 시간이 없어도, 순서 섹션의 나머지 5 시리즈를 배치하는 데이터를 보려는 그들이 웰스가 포함된 항원 보존 (1 % 폴리비닐 pyrrolidone, o.1M PBS에서 50 %의 에틸렌 글리콜, 수소 이온 농도 7.4)를 자르고 있었다. 아래 표는 1 슬라이드와 두뇌 은행 (표 1)의 일부로서 섹션 1 접시 (표준 48 잘 판)의 구성 샘플링 방식을 제공합니다. <img src="/files/ftp_upload/1260/table1.jpg" alt="표 1" /> 표 1. 여기서 섹션 샘플링 주파수 1 / 6입니다. 첫 번째 섹션은 cresyl 보라색 얼룩을 위해 슬라이드에 배치하고 향후 5 섹션은 보존 항원에 게재됩니다. 제 번호는 7 다음 슬라이드에서 캡처되고 섹션은 9-12 보존 항원에 배치됩니다. 관심이나 조직의 블록의 영역이 철저하게 sectioned 때까지이주기가 반복됩니다. </li><li> 다음 단계는 철저하게 섹션 조직의 블록을하는 것입니다. 척에 미디어를 삽입의 작은 금액을 놓고 그것이 동결하자. 마이크로톰 머리에 척을 놓고 평평한 표면을 가지고 냉동 매체의 정도 떨어져 면도. 마이크로톰 머리와 그라 이오 스탯 내에서 플랫 플레이스에서 척을 제거합니다. 척에 미디어를 삽입 붓고 단단히 척에 단호하게 위치 stereotaxically 컷 측면과 뇌 블록을 놓으십시오. 천천히 그리고 완전히는 매체를 장착의 머리를 포함. 마이크로톰 머리와 섹션 샘플링 주파수에 대해 미리 정해진 매개 변수를 사용하여 섹션으로 두뇌와 척를 놓습니다. </li><li> 조직의 블록이 철저하게 sectioned되었으며 섹션이 우물에 배치되면, 뚜껑과 접시를 커버 parafilm으로 뚜껑을 버리고, -20 ° C의 냉장고에서 개최. 각 시리즈, 섹션 - 샘플링 간격, 각 동물에 대한 일련의 번호를 촬영 섹션의 총 수를에 로그인합니다. 이 로그는 미래 immunohistochemistry를위한 두뇌 은행에서 섹션의 후속 제거를 추적할 중요한 것입니다. </li></ol> 4 부 : 대표 결과 : 이러한 방식으로 조직적인 샘플링은 지난 3 년간 우리 연구실에서 표준 연습을하고 있습니다. 그것이 신호의 열화 (그림 1)하지 않고 자른 후에 우리가 알게 된 지 3 년 보존 항원에 저장되었습니다 물질에 대한 성공적인 수행 immunohistochemistry의 좋은 거래를했다합니다. 또한, 우리의 두뇌 은행의 일환으로 우리가 아닌 인간의 가까이에 20,000 체계 섹션 로그인영장류의 두뇌 (그림 2) 장기 연구 계획의 일환으로. <img src="/files/ftp_upload/1260/1260figure1.jpg" alt="" /> 1.이 아닌 인간 영장류의 후두 극의 섹션입니다 그림 레이어 VI와 중간 흰색 물질 뉴런을 모두 보여주는 NeuN위한 immunostained. 이 특정 섹션 2 년간의 기간 동안 -20 ° C에 보존 항원에 저장되었습니다. 스케일 바 = 100μm. <img src="/files/ftp_upload/1260/1260fig2.jpg" alt="figure2" /> 그림 2. 우리 vervet 두뇌 은행은 현재 30여 원숭이에서 가까운 20,000 체계적인 섹션으로 구성되어 있습니다.

<ol>
	<li>West, M., Slomianka, L., Gundersen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unbiased+stereological+estimation+of+the+total+number+of+neurons+in+the+subdivisions+of+the+rat+hippocampus+using+the+optical+fractionator.+Aat+Rec.">Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator. Aat Rec.</a> 231, 482-497 (1991).</li><li>Burke, M. W., Zangenehpour, S., Boire, D., Ptito, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissecting+the+Non-human+Primate+Brain+in+Stereotaxic+Space.">Dissecting the Non-human Primate Brain in Stereotaxic Space.</a> J Vis Exp. , (2009).</li></ol>

The authors have nothing to disclose.

체계적인 샘플링 연구 자료를 극대화하기위한 저렴한 방법입니다. 이 샘플링 전략 지역에 걸쳐 체계적인 샘플링을 필요로 불편 stereology의 규칙을 준수하도록 설계되었습니다. 그것은 각 시리즈에 대한 섹션의 순서를 유지하는 것은 매우 중요합니다. 저희 연구실에서는 성공적으로 햄스터와 비 인간 영장류 두뇌 섹션을 모두 은행에 대해이 방법을 사용했습니다. 지금까지 우리는 가까이에 20,000 v...

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brain banking: making the most of your research specimens

불편 stereology 정확하고 효율적으로 관심 주어진 영역에서 총 뉴런 번호 (또는 다른 세포 유형) 계산하는 방법입니다<sup&gt; 1</sup&gt;. 이 목표를 달성하기 위해서는 6-10 체계 섹션은 전체 구조를 다루는 탐지해야합니다. 일반적으로이 처리되지 않은 물질의 상당한 양의 나뭇잎 5분의 1 섹션을 처리 포함됩니다. 자료를 극대화하기 위해, 우리는 장기 저장 연구 계획의 일환으로 고정 조직을 보존을위한 저렴한 방법을 제안합니다. 조직이 원하는 얼룩이나 항체, 다른 섹션에 대한 슬라이스 및 처리로가 체계적으로 미래를 처리 -20 ° C에 보존 항원에 배치해야합니다. 24 잘 접시를 사용하여 섹션 향후 검색을 위해 순서로 배열 수 있습니다. 이 방법을 사용하여 조직은 저장하고 년간의 과정을 통해 immunohistochemistry에 대해 처리할 수 있습니다.

Watch this Scientific Journal Video about Brain Banking: Making the Most of your Research Specimens at JoVE.com

Brain Banking: Making the Most of your Research Specimens

브레인 은행 및 생물 학적 물질의 체계적인 샘플링 불편 stereology의 기초를 제공하고 각 표본에서 얻은 잠재적인 데이터를 극대화할 수 있습니다.

브레인 은행 : 귀하의 연구 표본의 대부분을 만들기

Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given area of interest1.  To ...

brain-banking-making-the-most-of-your-research-specimens

Research

JoVE Journal

Biology

9.9K 조회수.  University of Montreal. 브레인 은행 및 생물 학적 물질의 체계적인 샘플링 불편 stereology의 기초를 제공하고 각 표본에서 얻은 잠재적인 데이터를 극대화할 수 있습니다.

비디오: Brain Banking: Making the Most of your Research Specimens

Micro-dissection of Rat Brain for RNA or Protein Extraction from Specific Brain Region

Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates micro-dissection of four major brain regions include olfactory bulb, frontal cortex, striatum and hippocampus in fresh rat brain tissue. Useful tips for quick removal of respective regions to avoid RNA and protein degradation of the tissue are given.

Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates micro-dissection of four major brain regions include olfactory bulb, frontal cortex, striatum and hippocampus in fresh rat brain tissue. Useful tips for quick removal of respective regions to avoid RNA and protein degradation of the tissue are given.

RNA 또는 특정 뇌 영역에서 단백질 추출에 대한 쥐 두뇌의 마이크로 해부

Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates ...

연구

생물학

Single Cell Electroporation in vivo within the Intact Developing Brain

Single-cell electroporation (SCE) is a specialized technique allowing the delivery of DNA or other macromolecules into individual cells within intact tissue, including in vivo preparations. The distinct advantage of this technique is that experimental manipulations may be performed on individual cells while leaving the surrounding tissue unaltered, thereby distinguishing cell-autonomous effects from those resulting from global treatments. When combined with advanced in vivo imaging techniques, SCE of fluorescent markers permits direct visualization of cellular morphology, cell growth, and intracellular events over timescales ranging from seconds to days. While this technique is used in a variety of in vivo and ex vivo preparations, we have optimized this technique for use in Xenopus laevis tadpoles. In this video article, we detail the procedure for SCE of a fluorescent dye or plasmid DNA into neurons within the intact brain of the albino Xenopus tadpole. We also discuss methods to optimize yield, and show examples of live two-photon fluorescence imaging of neurons fluorescently labeled by SCE.

Single-cell electroporation (SCE) is a specialized technique allowing delivery of DNA or other macromolecules into individual cells within intact tissue, including in vivo preparations. Here we detail the procedure for SCE of a fluorescent dye or plasmid DNA into neurons within the intact brain of the Xenopus laevis tadpole.

완전한 개발 두뇌 속에 생체내에서 단일 세포 Electroporation

Single-cell electroporation (SCE) is a specialized technique allowing the delivery of DNA or other macromolecules into individual cells within intact ...

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades. Today, such pipettes are made from glass capillaries using a programmable puller. Such instruments heat the capillary using either a metal filament or a laser and draw out the glass using gravity, a motor or both. Pipettes for patch-clamp recording are formed using only heat and gravity, while sharp electrodes for intracellular recording use a combination of heat, gravity, and a motor. The procedure used to make intracellular recording pipettes is similar to that used to make injection needles for a variety of applications, including cRNA injection into Xenopus oocytes. In general, capillary glass &lt;1.2 mm in diameter is used to make pipettes for patch clamp recording, while narrower glass is used for intracellular recording (outer diameter = 1.0 mm). For each tool, the puller is programmed slightly differently. This video shows how to make both kinds of recording pipettes using pre-established puller programs.

This video shows how to use a programmable puller to make patch pipettes and sharp electrodes for electrophysiology. The same procedure can be used to make a variety of glass tools, including injection needles.

프로그래밍 풀러와 패치 - pipettes와 샤프 전극을 만들기

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades.  Today, such pipettes are made from glass capillaries ...

Live Imaging of the Zebrafish Embryonic Brain by Confocal Microscopy

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the developing zebrafish brain (Gutzman et al, 2008). Such analysis affords a new understanding of the underlying cell biology required for development of the 3D structure of the vertebrate brain, and significantly increases our ability to study neural tube morphogenesis. The embryonic zebrafish brain is shaped beginning at 18 hours post fertilization (hpf) as the ventricles within the neuroepithelium inflate. By 24 hpf, the initial steps of neural tube morphogenesis are complete. Using the method described here, embryos at the one cell stage are injected with mRNA encoding membrane-targeted green fluorescent protein (memGFP). After injection and incubation, the embryo, now between 18 and 24 hpf, is mounted, inverted, in agarose and imaged by confocal microscopy. Notably, the zebrafish embryo is transparent making it an ideal system for fluorescent imaging. While our analyses have focused on the midbrain-hindbrain boundary and the hindbrain, this method could be extended for analysis of any region in the zebrafish to a depth of 
80-100 &mu;m.

In this video, we demonstrate a method by which to analyze the developing vertebrate brain in live zebrafish embryos at single cell resolution by confocal microscopy. This includes the method by which we inject the single-cell zebrafish embryo and subsequently mount and image the developing brain.

공촛점 현미경으로 Zebrafish 배아 뇌 영상 라이브

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the ...

Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural basis of human perception, cognition and behavior 1, 2. Moreover, this non-invasive imaging method has grown into a tool for clinicians and researchers to explore typical and atypical brain development. Although advances in neuroimaging tools and techniques are apparent, (f)MRI in young pediatric populations remains relatively infrequent 2. Practical as well as technical challenges when imaging children present clinicians and research teams with a unique set of problems 3, 2. To name just a few, the child participants are challenged by a need for motivation, alertness and cooperation. Anxiety may be an additional factor to be addressed. Researchers or clinicians need to consider time constraints, movement restriction, scanner background noise and unfamiliarity with the MR scanner environment2,4-10. A progressive use of functional and structural neuroimaging in younger age groups, however, could further add to our understanding of brain development. As an example, several research groups are currently working towards early detection of developmental disorders, potentially even before children present associated behavioral characteristics e.g.11. Various strategies and techniques have been reported as a means to ensure comfort and cooperation of young children during neuroimaging sessions. Play therapy 12, behavioral approaches 13, 14,15, 16-18 and simulation 19, the use of mock scanner areas 20,21, basic relaxation 22 and a combination of these techniques 23 have all been shown to improve the participant's compliance and thus MRI data quality. Even more importantly, these strategies have proven to increase the comfort of families and children involved 12. One of the main advances of such techniques for the clinical practice is the possibility of avoiding sedation or general anesthesia (GA) as a way to manage children's compliance during MR imaging sessions 19,20. In the current video report, we present a pediatric neuroimaging protocol with guidelines and procedures that have proven to be successful to date in young children.

Making MR Imaging Child&#039;s Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure

Despite an increase in the use of structural and functional magnetic resonance imaging (fMRI) in humans, the study of young pediatric populations remains a challenge. We present a hands-on, step-by-step video protocol including guidelines for clinicians and researchers intending to perform (f)MRI in young children.

소아 Neuroimaging 프로토콜, 지침 및 절차 - MR 이미징 아동의 놀이 만들기

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural ...

Making Gynogenetic Diploid Zebrafish by Early Pressure

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from heterozygous females allow F1 mutagenized females to be screened directly for deleterious mutations in an accelerated forward genetic screen. Streisinger et al.1,2 described methods for making gynogenetic (homozygous) diploid zebrafish by activating zebrafish eggs with ultraviolet light-inactivated sperm and preventing either the second meiotic or the first zygotic cell division using physical treatments (heat or pressure) that deploymerize microtubules. The "early pressure" (EP) method blocks the meiosis II, which occurs shortly after fertilization. The EP method produces a high percentage of viable embryos that can develop to fertile adults of either sex. The method generates embryos that are homozygous at all loci except those that were separated from their centromere by recombination during meiosis I. Homozygous mutations are detected in EP clutches at between 50% for centromeric loci and less than 1% for telomeric loci. This method is reproduced verbatim from the Zebrafish Book3.

This is a method for generating gynogenetic diploid zebrafish embryos (embryos whose only genetic contribution comes from the mother) by blocking the second meiotic division immediately after fertilization with ultraviolet light-inactivated sperm. EP embryos are not fully homozygous due to recombination during the first meiotic division, however they are homozygous at all loci that have not been separated from their centromere by recombination.

초기 압력에 의해 Gynogenetic의 Diploid의 Zebrafish 만들기

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from ...

Long-term Culture of Human Breast Cancer Specimens and Their Analysis Using Optical Projection Tomography

Breast cancer is a leading cause of mortality in the Western world. It is well established that the spread of breast cancer, first locally and later distally, is a major factor in patient prognosis. Experimental systems of breast cancer rely on cell lines usually derived from primary tumours or pleural effusions. Two major obstacles hinder this research: (i) some known sub-types of breast cancers (notably poor prognosis luminal B tumours) are not represented within current line collections; (ii) the influence of the tumour microenvironment is not usually taken into account.
We demonstrate a technique to culture primary breast cancer specimens of all sub-types. This is achieved by using three-dimensional (3D) culture system in which small pieces of tumour are embedded in soft rat collagen I cushions. Within 2-3 weeks, the tumour cells spread into the collagen and form various structures similar to those observed in human tumours1. Viable adipocytes, epithelial cells and fibroblasts within the original core were evident on histology. Malignant epithelial cells with squamoid morphology were demonstrated invading into the surrounding collagen. Nuclear pleomorphism was evident within these cells, along with mitotic figures and apoptotic bodies.
We have employed Optical Projection Tomography (OPT), a 3D imaging technology, in order to quantify the extent of tumour spread in culture. We have used OPT to measure the bulk volume of the tumour culture, a parameter routinely measured during the neo-adjuvant treatment of breast cancer patients to assess response to drug therapy. 
Here, we present an opportunity to culture human breast tumours without sub-type bias and quantify the spread of those ex vivo. This method could be used in the future to quantify drug sensitivity in original tumour. This may provide a more predictive model than currently used cell lines.

We have developed a collagen-based in vitro assay which promotes proliferation and invasion from samples of all breast cancer subtypes. Optical Projection Tomography, a three dimensional microscopy technique was utilised to visualise and quantify tumour expansion. This assay may be used to quantify drug response of individual tumour samples.

광학 프로젝션 Tomography를 사용하여 인간 유방암 표본과 그 분석의 장기 문화

Breast cancer is a leading cause of mortality in the Western world. It is well established that the spread of breast cancer, first locally and later ...

Obtaining Specimens with Slowed, Accelerated and Reversed Aging in the Honey Bee Model

Societies of highly social animals feature vast lifespan differences between closely related individuals. Among social insects, the honey bee is the best established model to study how plasticity in lifespan and aging is explained by social factors.
The worker caste of honey bees includes nurse bees, which tend the brood, and forager bees, which collect nectar and pollen. Previous work has shown that brain functions and flight performance senesce more rapidly in foragers than in nurses. However, brain functions can recover, when foragers revert back to nursing tasks. Such patterns of accelerated and reversed functional senescence are linked to changed metabolic resource levels, to alterations in protein abundance and to immune function. Vitellogenin, a yolk protein with adapted functions in hormonal control and cellular defense, may serve as a major regulatory element in a network that controls the different aging dynamics in workers.
Here we describe how the emergence of nurses and foragers can be monitored, and manipulated, including the reversal from typically short-lived foragers into longer-lived nurses. Our representative results show how individuals with similar chronological age differentiate into foragers and nurse bees under experimental conditions. We exemplify how behavioral reversal from foragers back to nurses can be validated. Last, we show how different cellular senescence can be assessed by measuring the accumulation of lipofuscin, a universal biomarker of senescence.
For studying mechanisms that may link social influences and aging plasticity, this protocol provides a standardized tool set to acquire relevant sample material, and to improve data comparability among future studies.

In honey bee workers, aging depends on social behaviors rather than on chronological age. Here we show how worker-types with very different aging patterns can be obtained and analyzed for cellular senescence.

둔화 가속과 반전 꿀 꿀벌 모델에서 노화와 표본을 얻기

Societies of highly social animals feature vast lifespan differences between closely related individuals. Among social insects, the honey bee is the best ...

Sampling Blood from the Lateral Tail Vein of the Rat

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, proteins, and glucose, yet the composition of the blood is dynamically regulated and easily perturbed. One factor that can change the blood composition is the stress response triggered by the sampling procedure, which can contribute to variability in the measures of interest. Here we describe a procedure for blood sampling from the lateral tail vein in the rat. This procedure offers significant advantages over other more commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, without anesthesia or analgesia. Additionally, it can be used to obtain large volume samples (upwards of 1 ml in some rats), and it may be used repeatedly across experimental days. By minimizing the stress response and pain resulting from blood sampling, measures can more accurately reflect the true basal state of the animal, with minimal influence from the sampling procedure itself.

Blood samples are useful for assessing biomarkers of physiological states or disease in vivo. Here we describe the methodology to sample blood from the lateral tail vein in the rat. This method provides rapid samples with minimal pain and invasiveness.

쥐의 측면 꼬리 정맥에서 혈액을 샘플링

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, ...

Adipo-Clear: A Tissue Clearing Method for Three-Dimensional Imaging of Adipose Tissue

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte precursors, immune cells, fibroblasts, blood vessels, and nerve projections. Although the molecular control of cell type specification and how these cells interact have been increasingly delineated, a more comprehensive understanding of these adipose-resident cells can be achieved by visualizing their distribution and architecture throughout the whole tissue. Existing immunohistochemistry and immunofluorescence approaches to analyze adipose histology rely on thin paraffin-embedded sections. However, thin sections capture only a small portion of tissue; as a result, the conclusions can be biased by what portion of tissue is analyzed. We have therefore developed an adipose tissue clearing technique, Adipo-Clear, to permit comprehensive three-dimensional visualization of molecular and cellular patterns in whole adipose tissues. Adipo-Clear was adapted from iDISCO/iDISCO+, with specific modifications made to completely remove the lipid stored in the tissue while preserving native tissue morphology. In combination with light-sheet fluorescence microscopy, we demonstrate here the use of the Adipo-Clear method to obtain high-resolution volumetric images&#160;of an entire adipose tissue.

Due to the high lipid content, adipose tissue has been challenging to visualize using traditional histological methods. Adipo-Clear is a tissue clearing technique that allows robust labeling and high-resolution volumetric fluorescent imaging of adipose tissue. Here, we describe the methods for sample preparation, pretreatment, staining, clearing, and mounting for imaging.

Adipo-지우기: 조직 지방 조직의 3 차원 이미징 방법 삭제

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte ...

Methods Article

브레인 은행 : 귀하의 연구 표본의 대부분을 만들기