Name: a sectioning, coring, and image processing guide for high-throughput cortical bone sample procurement and analysis for synchrotron micro-ct
Uploaded: 2020-06-12T14:00:00.000+00:00
Duration: 7 min 10 s
Description: Watch this Scientific Journal Video about A Sectioning, Coring, and Image Processing Guide for High-Throughput Cortical Bone Sample Procurement and Analysis for Synchrotron Micro-CT at JoVE.com

이 논문에 설명된 연구는 캐나다 혁신, 자연 과학 및 공학 연구 위원회, 서스 캐처 원 대학, 서스 캐처 원 정부, 서부 경제 다각화 캐나다, 캐나다 국립 연구위원회 및 캐나다 보건 연구소의 캐나다 혁신 재단이 지원하는 캐나다 광원의 BMIT 시설에서 수행되었습니다. 저자는 캐나다 광원의 빔 라인 과학자, 특히 아담 웹, 데니스 밀러, 세르게이 가질로프, 닝 주 스카이스캔 SRμCT 및 백색 빔 현미경 시스템의 설정 및 문제 해결에 도움을 주셔서 감사합니다. 우리는 또한 이 연구 결과에 대한 cadaveric 견본에 접근을 위한 Toledo 의과 대학의 대학 및 박사 제프리 웬스트루프에서 베스 달젤에게 감사드립니다. JM Andronowski는 애크런 대학과 형사 사법 목적 부여를위한 법의학 과학의 국립 사법 연구 개발 연구소 (2018-DU-BX-0188)에서 제공하는 창업 연구 기금을 통해 지원됩니다. RA 데이비스는 애크런 대학에서 제공하는 대학원 보조에 의해 지원됩니다. 코링 과 톱질에 사용되는 장비와 공급은 아크론 대학과 NSF가 CW 홀리오크에 EAR-1624242를 부여하는 스타트업 기금에 의해 구입되었습니다.

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저자는 공개 할 것이 없습니다.

제한된 FOV 설정을 갖춘 고해상도 SRμCT 이미징을 위한 균일한 및 원통형 피질 뼈 코어 샘플을 조달하기 위한 포괄적이고 표준화된 프로토콜은 없었습니다. 여기에 자세히 설명된 프로토콜은 SRμCT 이미징에 대한 일관되게 크기의 피질 뼈 코어 샘플을 조달하는 방법과 미세 건축 데이터의 후속 정확한 시각화 및 추출에 대한 포괄적인 자습서를 제공함으로써 무효화를 채웁니다. 당사의 프로토콜은 ?...

이미징 기술의 최근 발전으로, 지금 매우 높은 해상도의 X 선 이미징 데이터를 취득하는 것이 가능합니다. 데스크탑 마이크로 CT(μCT) 시스템은 비파괴적 특성1로인해 방수 골격을 이미징하는 현재 표준입니다. 그러나 피질 뼈의 미세 구조 적 특징을 이미징 할 때 μCT 사용은 더 제한적이었습니다. 해상도 제약으로 인해 데스크톱 시스템은 골세포 라쿠나에 같은 피질 모공보다 작은 미세 구조 적 특징을 이미지하는 데 필요한 해상도를 달성할 수 없습니다. 이 응용 프로그램의 경우 SRμCT는 이러한 시스템1의해상도가 커지므로 이상적입니다. 예를 들어, 생물 의학 이미징 및 치료 (BMIT) 빔 라인2에 캐나다 광원 (CLS)에서 실험은 0.9 μm만큼 작은 복셀이미지를 생산하고있다. 이전 연구1,3,4,5는 인간 긴 뼈(그림 1)에서피질 뼈 표본으로부터 프로젝션및 후속 3차원 (3D)을 획득하기 위해 이 해상도를 사용하여 골구세포 라부나 밀도4,6,7,9및라쿠나 모양및 크기3의 변화및 성간 사이의 변화. 추가 연구는 인간에서 골연 밴딩의 존재를입증했다 10,이전에 법의학 인류학 문헌에서 비 인간 포유동물과 연관된 것으로 인식 되는 현상.
뛰어난 해상도를 달성하기 위해 X선 빔은 최대 시편 크기를 직경 이내로 제한하는 시야(FOV) 내에 미세하게 초점을 맞추어야 합니다. 현재, 이러한 제한을 충족하는 뼈 샘플 조달을 요약하는 문헌에 설명된 포괄적이고 표준화된 절차는 없었습니다. FOV 내의 중심 표본은 1) 이미징 중에 180°를 회전할 때 샘플이 중심을 유지하고 2) 스캔 아티팩트가 이미지 잘림이 없기 때문에 제한되도록 하는 것이 중요합니다. 즉, FOV 외부의 시료의 어떤 부분도 FOV 내부의 초점으로 들어가는 빔을 방해하지 않는다. 이 경우 재구성 알고리즘은 완전히 올바른 재구성에 필요한 일부 감쇠 데이터가 박탈됩니다. 360° (전체 회전) 스캔빔 경화의 영향을 최소화하지만 이미징 중 정렬 불량 및 샘플 움직임으로 인한 아티팩트를 증가시키는 것은 주목할 가치가 있습니다. 따라서 360° 검사는 일반적으로 더 깨끗한 데이터를 생성하지만 이미징 시간이 두 배가되므로 실험 비용과 데이터 품질 간의 절충안을 해결해야 합니다.
뼈 화상 진찰 실험의 중요하고 수시로 간과되는 양상은 스캐닝의 앞에 수행된 정확하고 복제가능한 견본 준비 기술입니다. SRμCT 방법을 실험에 통합하는 연구는 샘플링 프로토콜을 간략하게 언급하지만 저자는 표본을 수집하는 데 사용되는 특정 방법론에 대해 거의 또는 전혀 세부 사항을 제공하지 않습니다. 많은 연구는 임의 차원의 직선 뼈 블록을 절단 언급하지만, 일반적으로사용되는도구 또는 포함 재료에 대한 자세한 정보를 제공하지3,4,10,11,12,13,14. 일부 연구자들은 일반적으로 핸드헬드 로터리 도구(예를 들어, Dremel)를 사용하여 관심 영역(ROI)3,4,10,11, 12,13,14에서뼈의 직선 블록을 제거한다. 이 방법은 FOV보다 더 큰 균일하게 크기의 샘플을 생성하여 아티팩트 및 이미지 잘림 스캔가능성을 높입니다. 이러한 표본은 종종 정밀 다이아몬드 웨이퍼 톱 (예 : Buehler Isomet)을 사용하여 추가 정제가 필요합니다. 일관된 차원(200/mm)으로 샘플을 조달하는 것은 획득된 데이터 집합이 최고 품질이고 후속 결과가 복제가능한지 확인하는 데 중요합니다.
샘플 조달 방법론의 제한된 보고는 이전 연구에서 수행된 방법을 사용 및/또는 검증하려고 할 때 어려움을 더합니다. 현재 연구원은 샘플링 절차에 대한 자세한 내용은 저자에게 직접 문의해야 합니다. 여기에 자세히 설명된 프로토콜은 생물 의학 연구자들이 철저히 문서화되고 복제되고 비용 효율적인 샘플링 기술을 제공합니다. 이 문서의 주요 목적은 미세 건축 데이터의 정확한 시각화 및 추출을 위해 밀 드릴 프레스 및 다이아몬드 코링 비트를 사용하여 일관되게 크기의 피질 뼈 코어 샘플을 조달하는 방법에 대한 포괄적 인 자습서를 제공하는 것입니다. 이 방법은 고압 암석 역학15,16,17,18,19의단단한 재료 블록에서 균일하고 작은 직경 (1-5 mm) 실린더를 일상적으로 수집하는 데 사용되는 절차에서 수정됩니다.

<table><tbody><tr><td>1-1/8&quot; plunge cutting carbide for composites</td><td>Warrior</td><td>61812</td><td>28.6mm plunge</td></tr><tr><td>70% Ethanol</td><td>Fisher Scientific</td><td>BP8201500</td><td>3.8 Liters</td></tr><tr><td>Blunt-tipped forceps</td><td>Fisher Scientific</td><td>10-300</td><td></td></tr><tr><td>Centrifuge tubes</td><td>ThermoFisher</td><td>55398</td><td></td></tr><tr><td>Crystalbond 509-3 Epoxy</td><td>Ted Pella</td><td>821-3</td><td></td></tr><tr><td>CTAnalyser</td><td>Bruker microCT</td><td>v.1.15.4.0</td><td>Download and install at https://www.bruker.com/products/microtomography/micro-ct-software/3dsuite.html</td></tr><tr><td>Dental Tool Kit</td><td>Amazon</td><td>787269885110</td><td></td></tr><tr><td>Diamond wafering saw blade for composite material</td><td>Buehler</td><td>#11-4247</td><td></td></tr><tr><td>Drill Press</td><td>Jet Mill/Drill</td><td>350017</td><td>Model: JMD-15, benchtop drill presses are suitable substites, but typically lack a translatable machine table for positioning samples beneath the drill stem</td></tr><tr><td>Fine-tipped forceps</td><td>Fisher Scientific</td><td>22-327379</td><td></td></tr><tr><td>Fixturing clamps for XY machine table for mill/drill</td><td>MSC Industrial Supply</td><td>#04804571</td><td></td></tr><tr><td>Glass microscope slides</td><td>Ted Pella</td><td>26005</td><td>75x50mm slides, 1mm thick</td></tr><tr><td>Glass slide chuck</td><td>Buehler</td><td>#112488</td><td>Large enough to hold 75x50mm glass slides</td></tr><tr><td>Hot plate capable of reaching 140 &amp;deg;C</td><td>ThermoScientific</td><td>HP88850105</td><td></td></tr><tr><td>Incubator</td><td>NAPCO</td><td>Model 4200</td><td></td></tr><tr><td>Isocut Fluid</td><td>Buehler</td><td>111193032</td><td>Lubricant; 30mL</td></tr><tr><td>Jeweler&#039;s diamond coring drill bit</td><td>Otto Frei</td><td>#119.050</td><td>2mm inner diameter hollow stem coring bit</td></tr><tr><td>NRecon</td><td>Bruker microCT</td><td>v.1.6.10.2</td><td>Download and install at https://www.bruker.com/products/microtomography.html</td></tr><tr><td>Oscillating saw</td><td>Harbor Freight</td><td>62866</td><td></td></tr><tr><td>Oven-safe glass dishes</td><td>Pyrex</td><td>1117715</td><td>Glass food storage container</td></tr><tr><td>Precision slow-speed saw (Isomet 1000)</td><td>Buehler</td><td>111280160</td><td></td></tr><tr><td>Razor blades</td><td>Amazon</td><td>25181</td><td></td></tr><tr><td>Shallow aluminum tins</td><td>Amazon</td><td>B01MRWLD0R</td><td>~8cm diameter</td></tr><tr><td>Specimen cups</td><td>Amazon</td><td>616784425436 885334344729</td><td></td></tr><tr><td>Tergazyme detergent</td><td>Alconox</td><td>1304-1</td><td>1.8kg box</td></tr><tr><td>Ultrasonic cleaner</td><td>MTI Corporation</td><td>KJ201508006</td><td></td></tr></tbody></table>

a sectioning, coring, and image processing guide for high-throughput cortical bone sample procurement and analysis for synchrotron micro-ct

뼈는 인간의 수명 동안 구조에 변화 하는 동적 및 기계적으로 활성 조직. 뼈 리모델링 과정의 제품은 기존의 2차원 기술을 사용하여 실질적으로 연구되었습니다. 데스크톱 마이크로 컴퓨팅 단층 촬영(μCT) 및 싱크로트론 방사선 마이크로 컴퓨터 단층 촬영(SRμCT)을 통한 X선 이미징 기술의 최근 발전은 다른 3D 이미징 기술(예를 들어, SEM)보다 더 큰 시야(FOV)의 고해상도 3차원(3D) 스캔을 획득하여 인간 내 마이크로코픽 구조의 보다 완전한 그림을 제공한다. 그러나 표본은 FOV 내에서 정확하게 중심을 두어 데이터 분석에 영향을 미치는 것으로 알려진 줄무늬 아티팩트의 모양을 제한해야 합니다. 이전 연구는 고르지 못한 가장자리 또는 이미지 잘림으로 인해 이미징 아티팩트를 초래하는 불규칙한 모양의 직선 골 블록의 조달을 보고했습니다. 우리는 인간 페모라의 전방 양으로부터 SRμCT 실험을 위해 일관되게 크기의 피질 뼈 코어 표본을 조달하기 위해 지질 학적 샘플링 프로토콜 (coring)을 적용했습니다. 이 코링 방법은 효율적이고 조직에 최소한의 파괴적입니다. 균일한 원통형 샘플을 생성하여 회전 시 동포메트릭이 되고 스캔 전반에 걸쳐 X선 빔에 대한 균일한 경로 길이를 제공하는 특성상 이미징 아티팩트를 감소시합니다. 코어및 불규칙한 모양의 샘플의 X선 지형 데이터의 이미지 처리는 피질 뼈 미세 아키텍처의 시각화 및 분석을 개선하는 기술의 잠재력을 확인합니다. 이 프로토콜의 목표는 다양한 유형의 고해상도 뼈 이미징 실험에 적응할 수 있는 피질 뼈 코어추출을 위한 신뢰할 수 있고 반복가능한 방법을 제공하는 것입니다. 이 작업의 가장 중요한 목표는 저렴하고 일관되며 간단하며 SRμCT를 위한 표준화된 피질 뼈 조달을 만드는 것입니다. 이 절차는 생물학적 인류학, 지구과학 또는 물질 과학과 같은 단단한 복합 물질을 일반적으로 평가하는 관련 분야의 연구원에 의해 더 적응될 수 있습니다.

설명된 코어 샘플링 방법은 매우 효과적이고 효율적인 것으로 입증되었습니다. 이 프로토콜을 사용하여 코링 시편은 CLS BMIT-BM 빔라인2의실험을 위해 일관되게 크기의 샘플을 조달할 수 있었으며, FOV는 1.49 μm 복셀 크기로 ~2mm입니다. 코어 직경의 일관성을 검증하기 위해 인간 전방 대퇴코어(n=69)의 하위 집합의 길이(위,중간, 아래)를 따라 세 가지 측정을 수행하였다. 코어?...

Watch this Scientific Journal Video about A Sectioning, Coring, and Image Processing Guide for High-Throughput Cortical Bone Sample Procurement and Analysis for Synchrotron Micro-CT at JoVE.com

A Sectioning, Coring, and Image Processing Guide for High-Throughput Cortical Bone Sample Procurement and Analysis for Synchrotron Micro-CT

우리는 인간 페모라의 전방 측면에서 SRμCT 실험을 위해 균일한 크기의 피질 뼈 표본을 조달하기 위해 지질학적(coring) 샘플링 프로토콜을 사용했습니다. 이 방법은 최소한의 파괴적이고 효율적이며, 불규칙한 샘플 모양에서 이미징 아티팩트를 최소화하고 미세 건축 시각화 및 분석을 향상시키는 원통형 표본을 초래합니다.

싱크로트론 마이크로 CT를 위한 고처리량 피질 뼈 샘플 조달 및 분석을 위한 단면, 코링 및 이미지 처리 가이드

Bone is a dynamic and mechanically active tissue that changes in structure over the human lifespan. The products of the bone remodeling process have been ...

a-sectioning-coring-image-processing-guide-for-high-throughput

Research

JoVE Journal

Biology

5.0K 조회수.  The University of Akron. 우리는 인간 페모라의 전방 측면에서 SRμCT 실험을 위해 균일한 크기의 피질 뼈 표본을 조달하기 위해 지질학적(coring) 샘플링 프로토콜을 사용했습니다. 이 방법은 최소한의 파괴적이고 효율적이며, 불규칙한 샘플 모양에서 이미징 아티팩트를 최소화하고 미세 건축 시각화 및 분석을 향상시키는 원통형 표본을 초래합니다.

비디오: A Sectioning, Coring, and Image Processing Guide for High-Throughput Cortical Bone Sample Procurement and Analysis for Synchrotron Micro-CT

High Resolution 3D Imaging of Ex-Vivo Biological Samples by Micro CT

Non-destructive volume visualization can be achieved only by tomographic techniques, of which the most efficient is the x-ray micro computerized tomography (&mu;CT).
High resolution &mu;CT is a very versatile yet accurate (1-2 microns of resolution) technique for 3D examination of ex-vivo biological samples1, 2. As opposed to electron tomography, the &mu;CT allows the examination of up to 4 cm thick samples. This technique requires only few hours of measurement as compared to weeks in histology. In addition, &mu;CT does not rely on 2D stereologic models, thus it may complement and in some cases can even replace histological methods3, 4, which are both time consuming and destructive. Sample conditioning and positioning in &mu;CT is straightforward and does not require high vacuum or low temperatures, which may adversely affect the structure. The sample is positioned and rotated 180&deg; or 360&deg;between a microfocused x-ray source and a detector, which includes a scintillator and an accurate CCD camera, For each angle a 2D image is taken, and then the entire volume is reconstructed using one of the different available algorithms5-7. The 3D resolution increases with the decrease of the rotation step. The present video protocol shows the main steps in preparation, immobilization and positioning of the sample followed by imaging at high resolution.

High Resolution 3D Imaging of Ex-Vivo Biological Samples by Micro CT

Non-destructive volume visualization can be achieved only by tomographic techniques, of which the most efficient is the x-ray micro computerized tomography ( CT).

고해상도 3D 영상의 전 VIVO</em마이크로 중부 표준시로> 생물 학적 샘플

Non-destructive volume visualization can be achieved only by tomographic techniques, of which the most efficient is the x-ray micro computerized ...

연구

생체공학

Automated Quantification of Hematopoietic Cell &#8211; Stromal Cell Interactions in Histological Images of Undecalcified Bone

Confocal microscopy is the method of choice for the analysis of localization of multiple cell types within complex tissues such as the bone marrow. However, the analysis and quantification of cellular localization is difficult, as in many cases it relies on manual counting, thus bearing the risk of introducing a rater-dependent bias and reducing interrater reliability. Moreover, it is often difficult to judge whether the co-localization between two cells results from random positioning, especially when cell types differ strongly in the frequency of their occurrence. Here, a method for unbiased quantification of cellular co-localization in the bone marrow is introduced. The protocol describes the sample preparation used to obtain histological sections of whole murine long bones including the bone marrow, as well as the staining protocol and the acquisition of high-resolution images. An analysis workflow spanning from the recognition of hematopoietic and non-hematopoietic cell types in 2-dimensional (2D) bone marrow images to the quantification of the direct contacts between those cells is presented. This also includes a neighborhood analysis, to obtain information about the cellular microenvironment surrounding a certain cell type. In order to evaluate whether co-localization of two cell types is the mere result of random cell positioning or reflects preferential associations between the cells, a simulation tool which is suitable for testing this hypothesis in the case of hematopoietic as well as stromal cells, is used. This approach is not limited to the bone marrow, and can be extended to other tissues to permit reproducible, quantitative analysis of histological data.

A strategy to quantitatively analyze histological data in the bone marrow is presented. Confocal microscopy of fluorescently labeled cells in tissue sections results in 2-dimensional images, which are automatically analyzed. Co-localization analyses of different cell types are compared to data from simulated images, giving quantitative information about cellular interactions.

조혈 세포의 자동화 된 정량화 - Undecalcified 뼈의 조직 학적 이미지에서 기질 세포 상호 작용

Confocal microscopy is the method of choice for the analysis of localization of multiple cell types within complex tissues such as the bone marrow. ...

발생학

Outer-Boundary Assisted Segmentation and Quantification of Trabecular Bones by an Imagej Plugin

Micro-computed tomography (micro-CT) is routinely used to assess bone quantity and trabecular microstructural properties in small animals under different bone loss conditions. However, the standard approach for trabecular analysis of micro-CT images is slice-by-slice semi-automatic hand-contouring, which is labor intensive and error prone. Described here is an efficient method for automatic segmentation of trabecular bones according to the bone's outer boundaries, where trabecular bones can be identified and segmented automatically with accuracy with less operator bias when appropriate segmentation parameters are set. To profile satisfactory segmentation parameters, an image stack of segmentation results is displayed, where all possible combinations of the segmentation parameters are changed one by one in sequence, and segmentation results with associated parameters can easily be visually checked. As a quality-control feature of the plugin, simulated standard objects are quantified where the measured quantities can be compared with theoretical values. Layer-by-layer quantification of trabecular properties and trabecular thicknesses are reported by such a plugin, and the distributions of such properties within the selected regions can be profiled easily. Although layer-by-layer quantification retains more information about trabecular bones and facilitates further statistical analysis of structural changes, such measures are unavailable from the output of current commercial software, where only a single quantified value for each parameter is reported for each sample. Therefore, the described workflows are better approaches for analyzing trabecular bones with accuracy and efficiency.

We present a workflow for segmenting and quantifying trabecular bones for 2D and 3D images based on the bone&#39;s outer boundary using an ImageJ plugin. This approach is more efficient and accurate than the current manual hand-contouring approach, and provides layer-by-layer quantifications, which are not available in current commercial software.

외부-경계 지원 세분화 및 정량화 Imagej 플러그인에 의해 배수 뼈의

Micro-computed tomography (micro-CT) is routinely used to assess bone quantity and trabecular microstructural properties in small animals under different ...

3D Imaging of Soft-Tissue Samples using an X-ray Specific Staining Method and Nanoscopic Computed Tomography

We demonstrate a laboratory-based method combining X-ray microCT and nanoCT with a specific X-ray stain, which targets the cell cytoplasm. The described protocol is easy to apply, fast and suitable for larger soft-tissue samples. The presented methodology enables the characterization of crucial tissue structures in three dimensions and is demonstrated on a whole mouse kidney. The multiscale approach allows to image the entire mouse kidney and supports the selection of further volumes of interest, which are acquired with higher resolutions ranging into the nanometer range. Thereby, soft-tissue morphology with a similar detail level as the corresponding histological light microscopy images is reproduced. Deeper insights into the 3D configuration of tissue structures are achieved without impeding further investigations through histological methods.

A protocol for 3D visualization of microscopic tissue structures by using an X-ray specific staining method designed for X-ray computed tomography is presented.

X선 특이적 염색 방법 및 나노스코픽 컴퓨터 단층 촬영을 사용한 연조직 시료의 3D 이미징

We demonstrate a laboratory-based method combining X-ray microCT and nanoCT with a specific X-ray stain, which targets the cell cytoplasm. The described ...

A Micro-CT-based Method for Characterizing Lesions and Locating Electrodes in Small Animal Brains

Lesion and electrode location verification are traditionally done via histological examination of stained brain slices, a time-consuming procedure that requires manual estimation. Here, we describe a simple, straightforward method for quantifying lesions and locating electrodes in the brain that is less laborious and yields more detailed results. Whole brains are stained with osmium tetroxide, embedded in resin, and imaged with a micro-CT scanner. The scans result in 3D digital volumes of the brains with resolutions and virtual section thicknesses dependent on the sample size (12&#8211;15 and 5&#8211;6 &#181;m&#160;per voxel for rat and zebra finch brains, respectively). Surface and deep lesions can be characterized, and single tetrodes, tetrode arrays, electrolytic lesions, and silicon probes can also be localized. Free and proprietary software allows experimenters to examine the sample volume from any plane and segment the volume manually or automatically. Because this method generates whole brain volume, lesions and electrodes can be quantified to a much higher degree than in current methods, which will help standardize comparisons within and across studies.

This article describes a straightforward method to prepare small animal brains for micro-CT imaging, in which lesions can be quantified and electrodes located with high precision in the context of the whole brain.

병 변 특성화 및 작은 동물 두뇌에 전극을 위치에 대 한 마이크로-CT 기반 방법

Lesion and electrode location verification are traditionally done via histological examination of stained brain slices, a time-consuming procedure that ...

신경과학

An Unbiased Approach of Sampling TEM Sections in Neuroscience

Investigations of the ultrastructural features of neurons and their synapses are only possible with electron microscopy. Especially for comparative studies of the changes in densities and distributions of such features, an unbiased sampling protocol is vital for reliable results. Here, we present a workflow for the image acquisition of brain samples. The workflow allows systematic uniform random sampling within a defined brain region, and the images can be analyzed using a disector. This technique is much faster than extensive examination of serial sections but still presents a feasible approach to estimate the densities and distributions of ultrastructure features. Before embedding, stained vibratome sections were used as a reference to identify the brain region under investigation, which helped speed up the overall specimen preparation process. This approach was used for comparative studies investigating the effect of an enriched-housing environment on several ultrastructural parameters in the mouse brain. Based on the successful use of the workflow, we adapted it for the purpose of elemental analysis of brain samples. We optimized the protocol in terms of the time of user-interaction. Automating all the time-consuming steps by compiling a script for the open source software SerialEM helps the user to focus on the main work of acquiring the elemental maps. As in the original workflow, we paid attention to the unbiased sampling approach to guarantee reliable results.

We introduce a novel workflow for electron microscopy investigations of brain tissue. The method allows the user to examine neuronal features in an unbiased fashion. For elemental analysis, we also present a script that automatizes most of the workflow for randomized sampling.

신경 과학에서 가장 섹션 샘플링의 공평한 접근

Investigations of the ultrastructural features of neurons and their synapses are only possible with electron microscopy. Especially for comparative ...

골절 감수성에 대한 기계적 검사의 원리와 프로토콜

Practical Considerations for the Design, Execution, and Interpretation of Studies Involving Whole-Bone Bending Tests of Rodent Bones

Skeletal fragility leading to fracture is an American public health crisis resulting in 1.5 million fractures each year and $18 billion in direct care costs. The ability to understand the mechanisms underlying bone disease and the response to treatment is not only desired, but critical. Mechanical testing of bone serves as a valuable technique for understanding and quantifying a bone&#39;s susceptibility to fracture. While this method appears simple to perform, inappropriate and inaccurate conclusions may be reached if governing assumptions and key steps are disregarded by the user. This has been observed across disciplines as studies continue to be published with misuse of methods and incorrect interpretation of results. This protocol will serve as a primer for the principles associated with mechanical testing along with the application of these techniques-from considerations of sample size through tissue harvesting and storage, to data analysis and interpretation. With this in hand, valuable information regarding a bone&#39;s susceptibility to fracture may be obtained, furthering understanding for both academic research and clinical solutions.

저자 스포트라이트: Enhancing Accuracy and Reproducibility in Whole Bone Bending Tests(전체 뼈 굽힘 검사의 정확성 및 재현성 향상) 

Mechanical testing of rodent bones is a valuable method for extracting information regarding a bone's susceptibility to fracture. Lacking proper practical understanding, the results may be overinterpreted or lack validity. This protocol will serve as a guide to ensure mechanical tests are performed accurately to provide valid and functional data.

설치류 뼈의 전체 뼈 굽힘 시험과 관련된 연구의 설계, 실행 및 해석에 대한 실용적인 고려 사항

Skeletal fragility leading to fracture is an American public health crisis resulting in 1.5 million fractures each year and $18 billion in direct care ...

마우스 신생아 뇌 Micro-CT 스캐닝 및 이미지 처리

Micro-CT Imaging and Morphometric Analysis of Mouse Neonatal Brains

Neuroimages are a valuable tool for studying brain morphology in experiments using animal models. Magnetic resonance imaging (MRI) has become the standard method for soft tissues, although its low spatial resolution poses some limits for small animals. Here, we describe a protocol for obtaining high-resolution three-dimensional (3D) information on mouse neonate brains and skulls using micro-computed tomography (micro-CT). The protocol includes those steps needed to dissect the samples, stain and scan the brain, and obtain morphometric measurements of the whole organ and regions of interest (ROIs). Image analysis includes the segmentation of structures and the digitization of point coordinates. In sum, this work shows that the combination of micro-CT and Lugol's solution as a contrast agent is a suitable alternative for imaging the perinatal brains of small animals. This imaging workflow has applications in developmental biology, biomedicine, and other sciences interested in assessing the effect of diverse genetic and environmental factors on brain development.

저자 스포트라이트: 생쥐 신생아 뇌의 고해상도 이미징 – Lugol&#39;s Solution Contrast Agent를 사용한 Micro-CT 프로토콜

This study describes the steps for obtaining high-resolution images of neonatal mouse brains by combining micro-computed tomography (micro-CT) and a contrast agent in ex vivo samples. We describe basic morphometric analyses to quantify brain size and shape in these images.

Micro-CT 영상 및 생쥐 신생아 뇌의 형태 분석

Neuroimages are a valuable tool for studying brain morphology in experiments using animal models. Magnetic resonance imaging (MRI) has become the standard ...

싱크로트론 마이크로 CT를 위한 고처리량 피질 뼈 샘플 조달 및 분석을 위한 단면, 코링 및 이미지 처리 가이드

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고해상도 3D 영상의 전 VIVO 생물 학적 샘플

조혈 세포의 자동화 된 정량화 - Undecalcified 뼈의 조직 학적 이미지에서 기질 세포 상호 작용

외부-경계 지원 세분화 및 정량화 Imagej 플러그인에 의해 배수 뼈의

X선 특이적 염색 방법 및 나노스코픽 컴퓨터 단층 촬영을 사용한 연조직 시료의 3D 이미징

병 변 특성화 및 작은 동물 두뇌에 전극을 위치에 대 한 마이크로-CT 기반 방법

신경 과학에서 가장 섹션 샘플링의 공평한 접근

설치류 뼈의 전체 뼈 굽힘 시험과 관련된 연구의 설계, 실행 및 해석에 대한 실용적인 고려 사항

설치류 뼈의 전체 뼈 굽힘 시험과 관련된 연구의 설계, 실행 및 해석에 대한 실용적인 고려 사항

설치류 뼈의 전체 뼈 굽힘 시험과 관련된 연구의 설계, 실행 및 해석에 대한 실용적인 고려 사항

Micro-CT 영상 및 생쥐 신생아 뇌의 형태 분석