-1::1
Simple Hit Counter
Skip to content

Products

Solutions

×
×
Sign In

CN

EN - EnglishCN - 简体中文DE - DeutschES - EspañolKR - 한국어IT - ItalianoFR - FrançaisPT - Português do BrasilPL - PolskiHE - עִבְרִיתRU - РусскийJA - 日本語TR - TürkçeAR - العربية
Sign In Start Free Trial

RESEARCH

JoVE Journal

Peer reviewed scientific video journal

Behavior
Biochemistry
Bioengineering
Biology
Cancer Research
Chemistry
Developmental Biology
View All
JoVE Encyclopedia of Experiments

Video encyclopedia of advanced research methods

Biological Techniques
Biology
Cancer Research
Immunology
Neuroscience
Microbiology
JoVE Visualize

Visualizing science through experiment videos

EDUCATION

JoVE Core

Video textbooks for undergraduate courses

Analytical Chemistry
Anatomy and Physiology
Biology
Calculus
Cell Biology
Chemistry
Civil Engineering
Electrical Engineering
View All
JoVE Science Education

Visual demonstrations of key scientific experiments

Advanced Biology
Basic Biology
Chemistry
View All
JoVE Lab Manual

Videos of experiments for undergraduate lab courses

Biology
Chemistry

BUSINESS

JoVE Business

Video textbooks for business education

Accounting
Finance
Macroeconomics
Marketing
Microeconomics

OTHERS

JoVE Quiz

Interactive video based quizzes for formative assessments

Authors

Teaching Faculty

Librarians

K12 Schools

Biopharma

Products

RESEARCH

JoVE Journal

Peer reviewed scientific video journal

JoVE Encyclopedia of Experiments

Video encyclopedia of advanced research methods

JoVE Visualize

Visualizing science through experiment videos

EDUCATION

JoVE Core

Video textbooks for undergraduates

JoVE Science Education

Visual demonstrations of key scientific experiments

JoVE Lab Manual

Videos of experiments for undergraduate lab courses

BUSINESS

JoVE Business

Video textbooks for business education

OTHERS

JoVE Quiz

Interactive video based quizzes for formative assessments

Solutions

Authors
Teaching Faculty
Librarians
K12 Schools
Biopharma

Language

zh_CN

EN

English

CN

简体中文

DE

Deutsch

ES

Español

KR

한국어

IT

Italiano

FR

Français

PT

Português do Brasil

PL

Polski

HE

עִבְרִית

RU

Русский

JA

日本語

TR

Türkçe

AR

العربية

    Menu

    JoVE Journal

    Behavior

    Biochemistry

    Bioengineering

    Biology

    Cancer Research

    Chemistry

    Developmental Biology

    Engineering

    Environment

    Genetics

    Immunology and Infection

    Medicine

    Neuroscience

    Menu

    JoVE Encyclopedia of Experiments

    Biological Techniques

    Biology

    Cancer Research

    Immunology

    Neuroscience

    Microbiology

    Menu

    JoVE Core

    Analytical Chemistry

    Anatomy and Physiology

    Biology

    Calculus

    Cell Biology

    Chemistry

    Civil Engineering

    Electrical Engineering

    Introduction to Psychology

    Mechanical Engineering

    Medical-Surgical Nursing

    View All

    Menu

    JoVE Science Education

    Advanced Biology

    Basic Biology

    Chemistry

    Clinical Skills

    Engineering

    Environmental Sciences

    Physics

    Psychology

    View All

    Menu

    JoVE Lab Manual

    Biology

    Chemistry

    Menu

    JoVE Business

    Accounting

    Finance

    Macroeconomics

    Marketing

    Microeconomics

Start Free Trial
Loading...
Home
JoVE Journal
Neuroscience
血管灌注固定的穆林肺的空气膨胀
血管灌注固定的穆林肺的空气膨胀
JoVE Journal
Neuroscience
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Neuroscience
Air-Inflation of Murine Lungs with Vascular Perfusion-Fixation

血管灌注固定的穆林肺的空气膨胀

Full Text
8,289 Views
07:19 min
February 2, 2021

DOI: 10.3791/62215-v

Stacey M. Thomas1, Joseph Bednarek2, William J. Janssen1, Patrick S. Hume1

1Division of Pulmonary, Sleep and Critical Care Medicine, Department of Medicine,National Jewish Health, 2Section of Pediatric Gastroenterology, Hepatology, and Nutrition,Children's Hospital Colorado

AI Banner

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article presents a method for preserving lung structures through an innovative air-inflation with vascular perfusion-fixation technique. This method allows researchers to maintain cell morphology and location within airways, alveoli, and interstitium for detailed structure-function analyses. The preserved samples are suitable for subsequent histologic studies.

Key Study Components

Area of Science

  • Neuroscience
  • Biology
  • Histological Analysis

Background

  • Traditional methods often compromise cell morphology during lung fixation.
  • This technique combines air inflation with vascular perfusion fixation to improve sample integrity.
  • Maintaining location of cells within lung architecture is critical for accurate analyses.

Purpose of Study

  • To develop a method preserving both structure and function of lung tissues.
  • To facilitate detailed histologic examination of lungs post-fixation.
  • To investigate inflammatory responses in lung tissues effectively.

Methods Used

  • The method involves air inflation of lungs combined with vascular perfusion fixation.
  • The biological model used is the euthanized mouse lung.
  • This technique involves precise surgical incisions and careful administration of fluids.
  • Key steps include setting up an air-inflation apparatus and accurately controlling pressure during fixation.
  • Importance is placed on the timing and order of inflation and fixative perfusion to prevent damage.

Main Results

  • The technique successfully preserves cell morphology and location in lung tissues.
  • Unlike traditional methods, few immune cells are lost in the airway lumen, allowing for more accurate assessments.
  • The method enhances visibility of inflammatory responses within perfused lung tissues.
  • Histological analysis demonstrates the viability of the samples for various imaging techniques.

Conclusions

  • This study enables better preservation of lung structures for detailed analyses.
  • It has significant implications for understanding lung pathology and responses to injury or infection.
  • Overall, the method enhances histological examination capabilities in lung research.

Frequently Asked Questions

What are the advantages of this air inflation and fixation method?
This method preserves cell morphology and positioning in lung structures better than traditional methods, allowing for more accurate functional analyses.
How is the mouse lung prepared for inflation and fixation?
Lungs are prepared by making incisions to access the trachea and heart, and through precise placement of a perfusion needle into the right ventricle.
What types of data can be obtained using this technique?
This method allows for histological analysis, including staining and immunohistochemistry to evaluate lung structure and immune response.
How can this method be applied to other studies?
This air-inflation and perfusion fixation technique can be adapted for other tissues where preservation of structural details is critical for analysis.
What are key considerations when using this technique?
Careful attention must be paid during the surgical preparation and the perfusion steps to avoid damaging the lung tissue.

呈现是一种空气膨胀的方法,通过血管灌注固定肺部,保留细胞在气道、肺泡和间歇性中的位置,用于结构功能分析。恒定的气道压力通过空气膨胀室保持,而固定气道则通过右心室喷灌。肺被处理为组织学研究。

这是一种廉价的方法,结合了空气膨胀和血管灌注固定的优势,以保存肺部进行结构和功能分析。这项技术的主要优点是保存细胞形态和肺空域内的位置。在将诱饵存根适配器放入气管和灌注针插入右心室时,应小心谨慎,以确保足够的通货膨胀和固定。

要设置充气装置,将水柱的注射器放入环架中,测量从动物平台到水柱 25 厘米的垂直高度。将水柱管的末端连接到空气室的塞孔上,并将空气室的女性诱饵中的管子连接到动物加工容器上的止流管。确认动物加工容器外部的气室盖和塞孔关闭,从水柱到空气膨胀室的管子上的止损孔打开。

然后将注射器装满水至25厘米大关。水会流过注射器和管子进入空气室。一旦压力被平衡,水就会停止流动。

为了使肺部为通货膨胀做好准备,在安乐死老鼠肋骨笼下的腹部壁上做一个横向切口,在臀部上方做第二个横向腹壁切口。沿着中线从内部切口切到高级切口,并用钝剪刀小心地切入隔膜的侧面。隔膜一被刺穿,肺就会塌陷。

沿着隔膜横向切割,打开胸腔,沿着胸骨从西福伊德过程切割到杂耍机腔,横向在肋骨笼上方,以充分暴露心脏和肺部。固定肋骨笼的两侧,在气管上方的颈部进行中线切口。去除气管周围的皮肤、肌肉、甲状腺和结缔组织,并使用弯曲的钳子在后气管下滑动两片缝合线。

使用一块缝合线将通货膨胀诱饵存根适配器保持到位,并使用 18 量表一英寸针在气管上打一个小孔。将一个20仪表的诱饵存根适配器插入孔中,并将缝合线系在气管周围,立即与诱导存根采用者进入孔的位置发生差。然后将动物转移到动物加工容器中,并将诱饵存根适配器连接到动物加工容器内部的女性诱饵上。

为了充气肺部,将附着在灌注装置管上的25个量表5/8英寸的针头放入心脏右心室,切开腹主动脉,让血液从心脏排出,促进香水通过肺部的流动。打开动物加工容器外部的塞子,在 25 厘米的水中充气 5 分钟,同时注意注射器中的水位不会迅速下降。在肺充膨胀的最后一分钟,打开渗透泵,流量为每分钟10毫升。

肝素溶液应从瓶子流经管子进入动物。在使肺部充气五分钟后,关闭泵,将香水从肝素切换到固定剂。将水柱注射器降低到 20 厘米。

当压力变化时,气泡在水柱内移动是正常的。检查注射器中的水位。它应该是在25厘米大关。

然后等待一分钟,让肺部从25至20厘米的水压下放气,然后以每分钟6.5毫升的流量重新启动泵,进行10至15分钟的血管固定灌注。要提取膨胀和固定的肺组织,请将第二根线紧紧系在气管周围,远距离到诱饵存根适配器,并从气管中取出诱饵存根适配器。从心脏取出针头,用钝剪刀切开结缔组织,后切至中性,将肺和心脏从胸腔中取出,避免刺穿肺部。

小心地从肺部取出心脏,在50毫升圆锥管中将肺放入20至25毫升的固定剂中。将支撑气管的缝合线拉过锥形管的开口,并通过盖子的螺纹固定缝合线。然后倒置管子,以确保浮力充气的肺在固定中保持完全淹没,并按照标准协议处理肺部进行组织学研究。

在解剖过程中输入隔膜时,胸膜空间的完整性将被废除,肺部应崩溃。应用25厘米的水压后,空气会通过气管进入肺部,并且应该很容易观察到通货膨胀。一旦肺部完全扩张,通货膨胀压力可以降低到20厘米,以保持肺部充气而不过度扩张。

气管连合和切除胸腔后,肺部应保持充气。如果肺在动物准备或肺提取过程中被刺穿,则可能发生通货紧缩。对充气肺部段的组织学分析表明,使用传统液体膨胀固定的组织气道流明中很少有免疫细胞。

相比之下,炎症细胞被保存在通过血管灌注与空气膨胀固定的组织空域内。在空气膨胀与大量固定后,肺组织可能嵌入正式组织分割和随后的分析通过组织学技术,包括染色,免疫造血术和免疫荧光成像。

View the full transcript and gain access to thousands of scientific videos

Sign In Start Free Trial

Explore More Videos

神经科学 第168期 空气膨胀 血管灌注固定 气道 肺球菌 间歇性 巨噬细胞 肺 结构功能

Related Videos

离体小鼠肺动脉瓣的压力固定:一种以封闭构象分离和化学固定小鼠肺动脉瓣的程序

04:19

离体小鼠肺动脉瓣的压力固定:一种以封闭构象分离和化学固定小鼠肺动脉瓣的程序

Related Videos

2.6K Views

使用穆林精密切肺片对肺动脉内下氧肺血管收缩进行视频形态分析

13:32

使用穆林精密切肺片对肺动脉内下氧肺血管收缩进行视频形态分析

Related Videos

11.3K Views

在离体血液灌注大鼠肺备量化单微血管通透性

07:22

在离体血液灌注大鼠肺备量化单微血管通透性

Related Videos

9.9K Views

灌输和固定方法有用的小鼠肺癌研究

06:57

灌输和固定方法有用的小鼠肺癌研究

Related Videos

29.4K Views

通过小鼠肺切除术和假体植入测量内部肺表面积的标准方法

08:46

通过小鼠肺切除术和假体植入测量内部肺表面积的标准方法

Related Videos

13.9K Views

小鼠肺气肿评价持续压力下的肺固定

05:48

小鼠肺气肿评价持续压力下的肺固定

Related Videos

9.8K Views

肿瘤病理学鼠肺的灌注和膨胀

04:24

肿瘤病理学鼠肺的灌注和膨胀

Related Videos

21.2K Views

用于微CT成像的成人和早期产后小鼠肺的血管铸造

09:00

用于微CT成像的成人和早期产后小鼠肺的血管铸造

Related Videos

8.2K Views

在合并臭氧和LPS诱导穆林急性肺损伤期间可视化肺细胞适应

14:48

在合并臭氧和LPS诱导穆林急性肺损伤期间可视化肺细胞适应

Related Videos

5.5K Views

建立用于大鼠循环死亡后肺的新型离体肺灌注系统

09:14

建立用于大鼠循环死亡后肺的新型离体肺灌注系统

Related Videos

939 Views

JoVE logo
Contact Us Recommend to Library
Research
  • JoVE Journal
  • JoVE Encyclopedia of Experiments
  • JoVE Visualize
Business
  • JoVE Business
Education
  • JoVE Core
  • JoVE Science Education
  • JoVE Lab Manual
  • JoVE Quizzes
Solutions
  • Authors
  • Teaching Faculty
  • Librarians
  • K12 Schools
  • Biopharma
About JoVE
  • Overview
  • Leadership
Others
  • JoVE Newsletters
  • JoVE Help Center
  • Blogs
  • JoVE Newsroom
  • Site Maps
Contact Us Recommend to Library
JoVE logo

Copyright © 2026 MyJoVE Corporation. All rights reserved

Privacy Terms of Use Policies
WeChat QR code