这个作品是通过从加拿大自然科学和工程研究委员会（RGPIN 238382-2011）和健康研究（MOP115076）的加拿大学院资助。希瑟·保罗是由加拿大自然科学和工程研究理事会研究生奖学金和艾伯塔创新的健康解决方案的奖学金支持。梅根·哈勒姆是由自然科学与工程研究理事会研究生奖学金，一个弗雷德里克·班廷和查尔斯最佳加拿大研究生奖学金，和亚省儿童医院研究所培训奖遗传学，儿童发展，健康的支持。

该协议被批准卡尔加里动物保健委员会的大学和符合该指南实验动物的护理和使用 。 从后代1.独立坝<ol><li>挤奶12之前至少5分钟分开她的后代的坝。 注:大坝可以挤奶可达5-6小时分离1,6,13后，但是分离的时间超过4小时，可能会改变乳成分14。而分离时间似乎没有影响奶源容积 12，它是表示，维持整个研究一致分离时间。乳成分在整个哺乳期15可能发生变化，因此，应努力保持奶源一致的一天。最大的牛奶生产建议将发生在哺乳期第14天12。 </li><li>使用变暖室，确保幼崽能够保持适当的身体temperatu重新没有他们的母亲的存在的挤奶过程的持续时间。 注:在研究下面列出，挤奶在断奶，当水坝约22周龄的后代21日龄进行，因此也没有变暖室使用。 </li></ol> 2.设置和准备<ol><li>收集需要的挤奶过程的所有材料。 注:所有材料可在材料和设备表中找到。 </li><li>放置一个加热垫板凳挤奶的地方将发生并覆盖垫在垫吸收板凳上。 </li><li>设置麻醉系统。确保系统有足够的氧气，并开始前异氟醚。附加麻醉剂掩模将用于初始麻醉诱导到机器。放置，将用于对附近如果比初始麻醉剂掩模不同麻醉维持掩模。 </li><li>附加将25g针1毫升注射器催产素注射用无菌技术。 </li><li>打开加热垫使母体体温挤奶过程期间保持。注:监视所述加热垫的温度，以确保垫不会变得太热并且造成烫伤。或者，使用热源，诸如外科加热表，可以设置到特定的温度。 </li></ol> 3.麻醉大坝使用异氟醚<ol><li>打开氧气罐，并打开流量1升（1000毫升）每分钟。打开异氟醚的流动，并设定为5％。注意:避免直接吸入麻醉剂，防止麻醉剂蒸气积聚。 </li><li>麻醉大坝。 </li><li>如果需要切换到维修面具，把大坝平卧于吸收板凳垫。缺乏踏板反射确认麻醉。 </li><li>降低的异氟烷至2-3％的流量为维持麻醉。持续监测大坝整个程序，以确保德不发生呼吸PRESSION。注意:一旦在麻醉状态下，坝的眼睛应该使用无菌的眼润滑剂，以防止眼睛变干或者成为划伤保护。 </li></ol> 4.注射催产素<ol><li>确保催产素（20 USP单位/毫升）还未到达其到期日期。催产素消毒的小瓶消毒酒精擦拭/酒精清洁垫。 </li><li>催产素入注射器的使用无菌技术，制定2 IU（0.1毫升）。使用一个新的针头和注射器将挤奶每个坝。 注:催产素剂量范围通常为单个注射1至5 IU 1,6,12,16。或者，4国际单位/公斤体重的剂量可以使用12。 2 IU单剂量可如果挤奶困难遇到重复一次。 </li><li>腹腔注射催产素。针插入与朝向头部的针指向腹部的右下象限，以一定的角度15-30°，约0.5厘米深。 </li><li>向后拉柱塞，以确保在注射前的负压。如果任何流体（血液，尿液，肠内容物， 等）被吸入注射器，取出针，并尝试在注射用新的针头和注射器。如果没有流体被吸入注射催产素和立即丢弃针头和注射器成生物危害容器。 </li><li>等待大约5-15分钟的催产素刺激放乳。 </li></ol> 5.准备挤奶遗址<ol><li>您可以选择其中的牛奶将被收集网站/奶嘴。乳可从任何奶嘴12被收集。 </li><li>轻轻取出周围与剪挤奶奶头的毛皮，毛皮可能造成困难样品收集由于牛奶芯吸。温柔 - 围绕乳头的皮肤非常敏感，可能是干的，因此很容易受到划痕和泪水。 </li><li>的奶嘴我灭菌不是必要的，但可选，清洁用温水将奶嘴的皮毛被删除后。因为多个站点可能需要的奶源至少准备两个站点。 注:如果牛奶分析包括微生物分析，乳头区，可能需要消毒用碘5。 </li></ol> 6.奶源<ol><li>轻轻挤压奶嘴的基础上，手动排出乳汁收集。 注:如果牛奶分析包括微生物分析，第一滴几滴牛奶应丢弃。 </li><li>收集奶液滴进入毛细管时，填充所述毛细管。可以容纳体积较大（ 例如，50微升）缓解这一进程毛细管。 注意:如果难以收集奶遇到，催产素的第二剂量可以施用。推荐剂量不超过4 IU总和。 </li><li>分配从毛细管的牛奶到由无菌的离心管动它被用来从乳头到微量离心管的侧吸取乳汁的管的端部 - 观察通过毛细作用被拉出的毛细管的牛奶。 注:"吹出"使用连接到1ml注射器18G的针头未吸入到管内的奶。 </li><li>连续监视大坝疼痛或呼吸抑制的迹象，并相应地调整异氟醚的流动。 </li><li>继续如在本节中所述，直到足够的牛奶已收集为所选奶分析收集奶。对于下面的creamatocrit和蛋白质浓度测定描述，收集0.25毫升牛奶。 注:使用不同的挤奶网站，如果放乳减慢或所选择的网站不排出乳汁充分。最多约2.5毫升每只动物乳汁可以收集17，或至多每奶头0.5毫升作者建议，根据麻醉总时间限制为约45-60分钟，邻ř挤奶的45分钟。 </li><li>收集乳成microhematocrit管用于从新鲜牛奶样品creamatocrit测量，并密封与粘土密封剂管的端部。标注microhematocrit管坝ID和直立存放牛奶样品。 </li><li>当挤奶完成后，关闭异氟烷和氧气的流量。从大坝取下面膜，继续监测大坝，直到清醒。建议如果没有充分意识到，大坝应在恢复期放在吸收板凳垫，而不是直接在笼子里的床上用品，防止被褥被吸入或恢复过程中刮伤大坝的眼睛。 注意:不要让水坝无人看管，直到它重新获得足够的意识，以保持胸骨recumbancy。 </li><li>如果没有进一步的分析是必需的，冻牛奶在-80℃。其他人已经提出，牛奶可以在-20℃18被储存长达3小时，在4℃或5个月。 </li></ol> 7. Creamatocrit测量<ol><li>通过计算乳creamatocrit（奶油的乳样品中的百分比）19估计在牛奶中的脂肪含量。 注:测量与人乳已显示，无论是新鲜或冷冻牛奶可以用于一个creamatocrit测量，然而鲜奶更高度相关（r = 0.92与R = 0.90）与脂浓度11。使用新鲜或冷冻牛奶creamatocrit测量应保持一致的整个研究中，作为解冻牛奶都与一个小的下降creamatocrit相关值11。 </li><li>对于鲜奶，收集乳汁从乳头到microhematocrit管的样品;填写至少¾满（约15-20微升）。另外，搅拌好后得出鲜牛奶从收集的样品放入毛细管。密封到底用粘土密封胶。 </li><li>将毛细管插入比容离心器，用密封端指向外，保证了离心机是平衡的。 </li><li>开始的血细胞比容旋转（120秒，13700 XG）。 注:旋转时间或速度可能因离心分离机所用的模式改变。 </li><li>从离心机中取出管后的自旋是完整的，进行测量来计算creamatocrit。仔细观察样品的分离成奶油层和透明层。 </li><li>测量和记录流体在管的总长度和的脂肪（霜）的长度使用卡尺或尺子层。 注:creamatocrit表示为奶样本19内的膏层的百分比，计算为（奶油层的长度/牛奶柱的总长度）×100（ 图1A）。计算从creamatocrit测量脂肪浓度和能量值如下:脂肪浓度（g / L）=（creamatocrit（％） - 0.59）/0.146（图1B）19;能量值（KC人/ L）= 290 +（66.8 * creamatocrit（％）（图1C）19。 </li></ol> 8.蛋白质浓度测定<ol><li>使用牛血清白蛋白作为蛋白标准，确定使用标准蛋白质测定，总乳蛋白质浓度如洛瑞蛋白质测定法6。 注:牛奶稀释可能需要的乳蛋白质测量落入测定法的标准曲线内。 </li></ol>

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作者有没有利益冲突披露。所有的动物实验均按照批准廉政公署的协议进行的。

Investigations into maternal milk components have increased as interest in early life development research rises. As the sole source of nutrition during the neonatal period, the bioactive compounds in milk are essential for ideal growth and development, especially in the context of intestinal and immune health3. The method presented here is a simple, non-invasive method of collecting milk from the lactating rat in amounts sufficient for downstream analysis, such as oligosaccharide profiling6. The me...

牛奶是营养新生哺乳动物，对于婴儿生长发育1,2-提供能量和营养的唯一来源。而乳主要由细胞脂质和蛋白 1，它还包含了过多了调节后代生命早期发展，包括酶，碳水化合物，激素，抗体，生长因子，细胞因子，外来体，微泡，和小RNA这样的生物活性化合物的作为微小RNA 1,2。孕妇奶粉在建立后代免疫和肠道健康3中的基础性作用，再加上证据表明，母乳喂养的婴儿不易受疾病2，突出了识别与疾病过程的早期生活有关的牛奶成分和所涉及的分子机制的重要性在他们的行动。发育大鼠是一种流行调查的各种营养，生理效果模型，并在早期的化学干预-生命的发展4。因此，大鼠乳汁的分析可以提供新的洞察孕产妇和后代的健康。 目前的科学进步，现在提供了越来越多的深入调查，对健康和疾病的特定的牛奶成分的影响的机会。例如，牛奶细菌型材测序阐明在婴儿肠道5早期肠集群自己的角色，乳寡糖的质谱分析已通过母亲饮食6，和微小RNA的深度测序分泌提供的洞察的乳寡糖概况的改变母乳的脂肪球突出于基因转录，代谢和免疫功能7可能角色。 大鼠模型代表在产妇的研究8,9中最常用的模式生物之一。一个优点是其短怀孕期和哺乳期的，仅approximat伊利21天每一个;因此，从怀孕哺乳的开始的总时间表示短时间内在能够产生有价值的数据。大鼠的较大尺寸相比小鼠，在奶源的背景下，可提供相对于牛奶和易于奶源体积显著优势;牛奶产量的小鼠，例如，似乎是依赖于总体重以产生更多的奶10较重的小鼠。 在这里，提供了手动收集的牛奶哺乳大鼠的一般描述。此协议需要最少的设备，非侵入性的，廉价的，并且可以用于收集奶足够量用于进一步下游分析。简言之，大坝用异氟烷麻醉，放乳由催产素的刺激，以及牛奶通过牛奶手册表达收集到毛细管。最后，如​​牛奶两个主要组成部分是脂肪和蛋白质，简要DESCRIPTIOÑ ​​估计使用creamatocrit测量11和使用标准蛋白质测定总蛋白质浓度的定量乳脂肪含量的呈现。

<table><tbody><tr><td>&lt;strong&gt;&lt;em&gt;Equipment - Milking&lt;/em&gt;&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>1 ml syringes</td><td>BD-Canada</td><td>309602</td><td></td></tr><tr><td>25 G needles</td><td>BD-Canada</td><td>305122</td><td></td></tr><tr><td>18 G needles</td><td>BD-Canada</td><td>305196</td><td></td></tr><tr><td>50 &amp;mu;l Microdispenser Capillary Tubes</td><td>Fisher Scientific</td><td>21-169D</td><td></td></tr><tr><td>Oxytocin (20 USP Units/ml)</td><td>Bimeda-MTC</td><td>1OXY015</td><td></td></tr><tr><td>PPC Vet Isoflurane Inhalation Anesthetic, 250 ml</td><td>Fresenius Kabi</td><td>M60302</td><td>Used on the order of a veterinarian</td></tr><tr><td>Sterile Alcohol Prep Pad</td><td>Dukal</td><td>853</td><td></td></tr><tr><td>Absorbent Bench Underpad</td><td>VWR</td><td>82020-845</td><td></td></tr><tr><td>Maxi-Therm Hyper/Hypothermia Blanket</td><td>Cincinnati Sub-Zero</td><td>274</td><td></td></tr><tr><td>Rodent Anesthesia Machine with Vaporizer</td><td>Benson Medical Industries Inc.</td><td></td><td>Subject to individual laboratory needs</td></tr><tr><td>Animal Masks</td><td>Benson Medical Industries Inc.</td><td>50100/50102</td><td></td></tr><tr><td>Microcentrifuge Tubes</td><td>Axygen</td><td>MCT-060-C</td><td></td></tr><tr><td>ChroMini Professional Trimmer</td><td>Wahl</td><td></td><td></td></tr><tr><td>&lt;strong&gt;&lt;em&gt;Equipment - Creamatocrit&lt;/em&gt;&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>StatSpin SafeCrit Plastic Microhematocrit Tubes (Untreated)</td><td>Fisher Scientific</td><td>22-274-914</td><td></td></tr><tr><td>Critoseal Capillary Tube Sealant Tray</td><td>VWR</td><td>470161-478</td><td></td></tr><tr><td>StatSpin CritSpin Microhematocrit Centrifuge</td><td>Beckman Coulter, Inc</td><td>X00-004999-001</td><td></td></tr></tbody></table>

milk collection in the rat using capillary tubes and estimation of milk fat content by creamatocrit

Milk, as the sole source of nutrition for the newborn mammal, provides the necessary nutrients and energy for offspring growth and development. It also contains a vast number of bioactive compounds that greatly affect the development of the neonate. The analysis of milk components will help elucidate key factors that link maternal metabolism and health with offspring growth and development. The laboratory rat represents a popular model organism for maternal studies, and rat milk can be used to examine the effect of various maternal physiological, nutritional, and pharmacological interventions on milk components, which may then impact offspring health. Here a simple method of manually collecting milk from the lactating rat that can be performed by a single investigator, does not require specialized vacuum or suction equipment, and provides sufficient milk for subsequent downstream analysis is described. A method for estimating the fat content of milk by measuring the percentage of cream within the milk sample, known as the creamatocrit, is also presented. These methods can ultimately be used to increase insight into maternal-child health and to elucidate maternal factors that are involved in proper growth and development of offspring.

截至取Wistar水坝断奶描述的牛奶收集（约22周龄，体重350至400g），其消耗的控制（AIN-93G中，n = 5），高蛋白质（40％酪蛋白重量/重量，N = 5）或高益生纤维（21.6％重量/重量，1:1的比例的寡果糖和菊粉中，n = 4）的饮食整个怀孕和哺乳期。催产素剂量为2 IU。使用毛细管奶收集，并用血细胞比容喷丝确定creamatocrit（图1A），然后将其用于估计根据脂肪浓度和能量值一个管进行纺丝:脂肪浓?...

Watch this Scientific Journal Video about Milk Collection in the Rat Using Capillary Tubes and Estimation of Milk Fat Content by Creamatocrit at JoVE.com

Milk Collection in the Rat Using Capillary Tubes and Estimation of Milk Fat Content by Creamatocrit

Milk is a primary source of nutrition for the neonate. Analysis of milk components may provide insight into maternal factors that affect offspring health. This protocol describes a manual method of collecting milk samples from the lactating rat, which can then be used for further downstream analysis.

奶源在大鼠毛细管管和乳脂肪含量Creamatocrit估计

Milk, as the sole source of nutrition for the newborn mammal, provides the necessary nutrients and energy for offspring growth and development. It also ...

milk-collection-rat-using-capillary-tubes-estimation-milk-fat-content

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

12.3K 次观看.  University of Calgary. Milk is a primary source of nutrition for the neonate. Analysis of milk components may provide insight into maternal factors that affect offspring health. This protocol describes a manual method of collecting milk samples from the lactating rat, which can then be used for further downstream analysis.

视频: Milk Collection in the Rat Using Capillary Tubes and Estimation of Milk Fat Content by Creamatocrit

The Use of Gas Chromatography to Analyze Compositional Changes of Fatty Acids in Rat Liver Tissue during Pregnancy

Gas chromatography (GC) is a highly sensitive method used to identify and quantify the fatty acid content of lipids from tissues, cells, and plasma/serum, yielding results with high accuracy and high reproducibility. In metabolic and nutrition studies GC allows assessment of changes in fatty acid concentrations following interventions or during changes in physiological state such as pregnancy. Solid phase extraction (SPE) using aminopropyl silica cartridges allows separation of the major lipid classes including triacylglycerols, different phospholipids, and cholesteryl esters (CE). GC combined with SPE was used to analyze the changes in fatty acid composition of the CE fraction in the livers of virgin and pregnant rats that had been fed various high and low fat diets. There are significant diet/pregnancy interaction effects upon the omega-3 and omega-6 fatty acid content of liver CE, indicating that pregnant females have a different response to dietary manipulation than is seen among virgin females.

Pregnancy leads to significant changes to the fatty acid composition of maternal tissues. Lipid profiles can be obtained via gas chromatography to allow identification and quantification of fatty acids in individual lipid classes among rats fed various high and low fat diets during pregnancy.

使用气相色谱怀孕期间分析脂肪酸组成的变化在大鼠肝组织

Gas chromatography (GC) is a highly sensitive method used to identify and quantify the fatty acid content of lipids from tissues, cells, and plasma/serum, ...

科研

化学

Rat Mammary Epithelial Cell Transplantation into the Interscapular White Fat Pad

As early as the 1970s, researchers have successfully transplanted mammary epithelial cells into the interscapular white fat pad of rats. Grafting mammary epithelium using transplantation techniques takes advantage of the hormonal environment provided by the adolescent rodent host. These studies are ideally suited to explore the impact of various biological manipulations on mammary gland development and dissect many aspects of mammary gland biology. A common, but limiting, feature is that transplanted epithelial cells are strongly influenced by the surrounding stroma and outcompeted by endogenous epithelium; to utilize native mammary tissue, the abdominal-inguinal white fat pad must be cleared to remove host mammary epithelium prior to the transplantation. A major obstacle when using the rat model organism is that clearing the developing mammary tree in post-weaned rats is not efficient. When transplanted into gland-free fat pads, donor epithelial cells can repopulate the cleared host fat pad and form a functional mammary gland. The interscapular fat pad is an alternative location for these grafts. A major advantage is that it lacks ductal structures yet provides the normal stroma that is necessary to promote epithelial outgrowth and is easily accessible in the rat. Another major advantage of this technique is that it is minimally invasive, because it eliminates the need to cauterize and remove the growing endogenous mammary tree. Additionally, the interscapular fat pad contains a medial blood vessel that can be used to separate sites for grafting. Because the endogenous glands remain intact, this technique can also be used for studies comparing the endogenous mammary gland to the transplanted gland. This paper describes the method of mammary epithelial cell transplantation into the interscapular white fat pad of rats.

This article describes a transplantation method to graft donor rat mammary epithelial cells into the interscapular white fat pad of recipient animals. This method can be used to examine host and/or donor effects on mammary epithelium development and eliminates the need for pre-clearing, thereby extending the usefulness of this technique.

大鼠乳腺内皮细胞移植到间膜白脂肪垫

As early as the 1970s, researchers have successfully transplanted mammary epithelial cells into the interscapular white fat pad of rats. Grafting mammary ...

癌症研究

Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain

This study was designed to evaluate the protective effect of the Bacillus subtilis strain against complications related to heat stress. Thirty-two Sprague-Dawley rats were used in this study. Animals were orally treated twice a day for two days with B. subtilis BSB3 strain or PBS. The next day after the last treatment, each group was divided and two experimental groups (one treated with PBS and one treated with B. subtilis) were placed at 45 oC for 25 min. Two control groups stayed for 25 min at room temperature. All rats were euthanized and different parameters were analyzed in all groups. Adverse effects of heat stress are registered by the decrease of villi height and total mucosal thickness in the intestinal epithelium; translocation of bacteria from the lumen; increased vesiculation of erythrocytes and elevation of the lipopolysaccharides (LPS) level in the blood. The protective efficacy of treatment is evaluated by prevention of these side effects. The protocol was set up for the oral treatment of rats with bacteria for prevention of heat stress complications, but this protocol can be modified and used for other routes of administration and for analysis of different compounds.

Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain

We described a protocol for prevention of heat stress effects in rats by oral pre-treatment with beneficial bacteria. This protocol can be modified and used for various routes of administration and for analysis of different compounds.

热火预防应激在大鼠不利影响<em&gt;枯草芽孢杆菌</em&gt;应变

This study was designed to evaluate the protective effect of the Bacillus subtilis strain against complications related to heat stress. Thirty-two ...

免疫与感染

Repeated Blood Collection from Tail Vein of Non-Anesthetized Rats with a Vacuum Blood Collection System

Blood can be collected from rats in a number of sampling locations. For instance, the tail vein is a superior location for blood sampling. However, the tail vein is thin so that it is sometimes hard to puncture. In addition, the tail vein has low blood flow and requires a long sampling time to get sufficient blood. The present report describes a simple blood sampling method, the vacuum blood collection method, which is usually used to obtain blood samples from patients, here used for non-anesthetized rats. The 22 G butterfly needle tip was inserted into one of the lateral tail veins approximately 2-3 cm from the tip of the tail at an angle of approximately 20&#176;, and blood was collected in the vacuum collection tube by inserting the rubber end of the butterfly needle into the vacuum blood collection tube. The present experimental results show that the success rate was 95% in the experimental group and 90% in the beginner group. The success rate and puncture times were similar between two groups. The sampling duration was significantly shorter in the experimental group compared to beginner group. In conclusion, this vacuum blood collection method for sequential blood sampling from the tail vein of non- anesthetized rats is feasible and easy-to-learn, which might serve as a reliable alternative to other conventionally used blood sampling techniques for rats.

Here, we describe a simple tail vein blood sampling method in non-anesthetized rats using a vacuum extraction tube system. This method reduces the risk of direct exposure to blood and simplifies taking multiple samples from a single venipuncture.

无麻醉大鼠尾静脉血采集系统的研究

Blood can be collected from rats in a number of sampling locations. For instance, the tail vein is a superior location for blood sampling. However, the ...

医学

Three-Dimensional Culture of Vascularized Thermogenic Adipose Tissue from Microvascular Fragments

Engineering thermogenic adipose tissue (e.g., beige or brown adipose tissues) has been investigated as a potential therapy for metabolic diseases or for the design of personalized microtissues for health screening and drug testing. Current strategies are often quite complex and fail to accurately fully depict the multicellular and functional properties of thermogenic adipose tissue. Microvascular fragments, small intact microvessels comprised of arteriole, venules, and capillaries isolated from adipose tissue, serve as a single autologous source of cells that enable vascularization and adipose tissue formation. This article describes methods for optimizing culture conditions to enable the generation of three-dimensional, vascularized, and functional thermogenic adipose tissues from microvascular fragments, including protocols for isolating microvascular fragments from adipose tissue and culture conditions. Additionally, best practices are discussed, as are techniques for characterizing the engineered tissues, and sample results from both rodent and human microvascular fragments are provided. This approach has the potential to be utilized for the understanding and development of treatments for obesity and metabolic disease.

Here, we present a detailed protocol outlining the use of microvascular fragments isolated from rodent or human fat tissue as a straightforward approach to engineer functional, vascularized beige adipose tissue.

微血管碎片血管化产热脂肪组织的三维培养

Engineering thermogenic adipose tissue (e.g., beige or brown adipose tissues) has been investigated as a potential therapy for metabolic diseases or for ...

生物工程

Intranasal Immunization and Milk Collection in Studies of Maternal Immunization in New Zealand White Rabbits (Oryctolagus cuniculus)

Due to similarities in placentation and antibody transfer with humans, rabbits are an excellent model of maternal immunization. Additional advantages of this research model are the ease of breeding and sample collection, relatively short gestation period, and large litter sizes. Commonly assessed routes of immunization include subcutaneous, intramuscular, intranasal, and intradermal. Nonterminal sample collection for the chronological detection of the immunologic responses to these immunizations include the collection of blood, from both dams and kits, and milk from the lactating does. In this article, we will demonstrate techniques our lab has utilized in studies of maternal immunization in New Zealand White rabbits (Oryctolagus cuniculus), including intranasal immunization and milk collection.

Intranasal Immunization and Milk Collection in Studies of Maternal Immunization in New Zealand White Rabbits Oryctolagus cuniculus

This article describes and demonstrates the administration of intranasal vaccines and the collection of milk from lactating rabbits (Oryctolagus cuniculus) as a means to assess mucosal immunity in a translationally appropriate model of maternal immunization.

新西兰白兔产妇免疫研究中的内在免疫和牛奶收集

Due to similarities in placentation and antibody transfer with humans, rabbits are an excellent model of maternal immunization. Additional advantages of ...

Milk Collection Methods for Mice and Reeves' Muntjac Deer

Animal models are commonly used throughout research laboratories to accomplish what would normally be considered impractical in a pathogen&#8217;s native host. Milk collection from animals allows scientists the opportunity to study many aspects of reproduction including vertical transmission, passive immunity, mammary gland biology, and lactation. Obtaining adequate volumes of milk for these studies is a challenging task, especially from small animal models. Here we illustrate an inexpensive and facile method for milk collection in mice and Reeves&#8217; muntjac deer that does not require specialized equipment or extensive training. This particular method requires two researchers: one to express the milk and to stabilize the animal, and one to collect the milk in an appropriate container from either a Muntjac or mouse model. The mouse model also requires the use of a P-200 pipetman and corresponding pipette tips. While this method is low cost and relatively easy to perform, researchers should be advised that anesthetizing the animal is required for optimal milk collection.

Milk collection from animal models facilitates various research avenues: understanding passive immunity, identifying pathogens responsible for vertical transmission and, through the use of transgenic mice, even commercial production of proteins found in human breast milk. Here we illustrate a simple method for milk collection in mice and Reeves&#8217; muntjac deer.

牛奶收集方法小鼠和里夫斯&#39;麂鹿

Animal models are commonly used throughout research laboratories to accomplish what would normally be considered impractical in a pathogen&#8217;s native ...

生物学

Preparation of Rat Serum Suitable for Mammalian Whole Embryo Culture

Mammalian whole embryo culture (WEC) is a widely used technique for examining pharmacological toxicity in developing mouse and rat embryos and for investigating the mechanisms of developmental processes. Immediately centrifuged (IC) rat serum is commonly used for WEC and is essential for the growth and development of cultured mouse and rat embryos ex vivo. For the culture of midgestation embryos (i.e., E8.0-12.5 for the mouse, and E10.0-14.5 for the rat), 100% rat serum is the best media for supporting the growth of the embryo ex vivo. To prepare rat serum suitable for WEC, the collected blood should be centrifuged immediately to separate the blood cells from the plasma fraction. After centrifugation, the fibrin clot forms in the upper layer; this clot should be squeezed gently using a pair of sterile forceps and subsequently centrifuged to completely separate the blood cells from the serum. In this video article, we demonstrate our standard protocol for the preparation of optimal IC rat serum, including blood collection from the abdominal aorta of male rats and extraction of the serum by centrifugation.

Mammalian whole embryo culture (WEC) is widely used in teratology and developmental biology. Immediately centrifuged rat serum is commonly provided as a medium for both mouse and rat WEC. In this video, we demonstrate our standard protocol for the preparation of high-quality rat serum suitable for mammalian WEC.

大鼠血清适用于哺乳动物全胚胎培养的制备

Mammalian whole embryo culture (WEC) is a widely used technique for examining pharmacological toxicity in developing mouse and rat embryos and for ...

同时记录脑电图的大鼠脑脊液和血液

Sampling Cerebrospinal Fluid and Blood from Lateral Tail Vein in Rats During EEG Recordings

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many experimental contexts to measure molecules of interest, such as hormones, growth factors, proteins, or small non-coding RNAs. A specific example is the sampling of biological liquids in the research of biomarkers for epilepsy. In these studies, it is desirable to compare the levels of molecules in cerebrospinal fluid (CSF) and in plasma, by withdrawing CSF and plasma in parallel and considering the time distance of the sampling from and to seizures. The combined CSF and plasma sampling, coupled with video-EEG monitoring in epileptic animals, is a promising approach for the validation of putative diagnostic and prognostic biomarkers. Here, a procedure of combined CSF withdrawal from cisterna magna and blood sampling from the lateral tail vein in epileptic rats that are continuously video-EEG monitored is described. This procedure offers significant advantages over other commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, and reduced time of anesthesia. Additionally, it can be used to obtain CSF and plasma samples in both tethered and telemetry EEG recorded rats, and it may be used repeatedly across multiple days of experiment. By minimizing the stress due to sampling by shortening isoflurane anesthesia, measures are expected to reflect more accurately the true levels of investigated molecules in biofluids. Depending on the availability of an appropriate analytical assay, this technique may be used to measure the levels of multiple, different molecules while performing EEG recording at the same time.

作者聚焦:获取高质量的 CSF 和血液样本以发现癫痫生物标志物 

The protocol shows repeated cerebrospinal fluid and blood collections from epileptic rats performed in parallel with continuous video-electroencephalogram (EEG) monitoring. These are instrumental for exploring possible links between changes in various body fluid molecules and seizure activity.

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many ...

奶源在大鼠毛细管管和乳脂肪含量Creamatocrit估计

摘要

探索更多视频

此视频中的章节

使用气相色谱怀孕期间分析脂肪酸组成的变化在大鼠肝组织

大鼠乳腺内皮细胞移植到间膜白脂肪垫

热火预防应激在大鼠不利影响

无麻醉大鼠尾静脉血采集系统的研究

微血管碎片血管化产热脂肪组织的三维培养

新西兰白兔产妇免疫研究中的内在免疫和牛奶收集

牛奶收集方法小鼠和里夫斯'麂鹿

大鼠血清适用于哺乳动物全胚胎培养的制备

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液