이 연구는 장쑤 고등 교육 기관의 우선 학업 프로그램 개발(PAPD) 및 중국 약학 대학의 교육 개혁 연구 프로젝트(2019XJYB18)의 지원을 받았습니다.

1. 반고체 배지의 제조
<ol><li>전통적인 반고체 한천
	<ol><li>기본 성분11을 사용하여 박테리아 운동성 시험 배지 레시피에 따라 전통적인 반고체 한천을 준비한다. 충분한 증류수에 트립토오스 10g, NaCl 15g, 한천 4g을 녹이고 pH를 7.2 ± 0.2로 조정하고 최종 부피를 1,000mL로 구성합니다.</li>
		<li>한천을 121°C에서 20분 동안 오토클레이브하고 3cm 높이의 반고체 배지로 10mL 시험관에 분주합니다.</li>
	</ol></li>
	<li>TTC를 사용한 전통적인 반고체 한천
	<ol><li>종래의 반고체 배지를 오토클레이빙한 후 50°C로 냉각하고 100mL의 배지에 멸균된 1% TTC 용액 5mL를 넣고 혼합한 후 10mL 시험관에 분주하여 3cm 높이의 반고체 배지를 형성합니다.</li>
	</ol></li>
</ol>2. 박테리아 균주
참고: 대장균, 녹농균, 살모넬라종, 비브리오종, 폐렴간균 및 에어로모나스 하이드로필라를 포함한 80개의 균주를 수생 환경에서 분리하고 자동 박테리아 식별 기기(재료 표 참조)를 사용하여 식별했습니다(표 1). 황색포도상구균(Staphylococcus aureus)(재료 표 참조)은 음성 비운동성 대조군으로 사용되었습니다. 대장균(Escherichia coli), 녹농균(Pseudomonas aeruginosa) 및 살모넬라 티피뮤리움(Salmonella typhimurium)(재료 표 참조)을 양성 대조군 균주로 사용했습니다.
<ol><li>운동성 분석에 사용할 테스트 박테리아 균주를 식별합니다.</li>
	<li>음성 비운동성 대조군 및 운동성 양성 대조군 균주를 포함합니다.</li>
</ol>3. TTC 강화 박테리아 운동성 관찰
<ol><li>한천 플레이트에서 테스트 박테리아의 단일 콜로니를 선택하고 접종 바늘을 사용하여 구멍을 뚫어 위의 두 반고체 배지(단계 1.1.2 및 1.2.1)에 접종합니다.</li>
	<li>튜브를 37°C에서 24-48시간 동안 배양하여 결과를 관찰하였다.</li>
	<li>성장 상태 관찰: 펑크 라인만 빨간색인 경우 박테리아를 운동성이 없는(-) 특성화합니다. 붉은 색이 천공선12를 따라 바깥쪽으로 가볍게 퍼지면 박테리아를 운동성(+)으로 특성화합니다.</li>
</ol>4. 다양한 한천 농도가 박테리아 운동성에 미치는 영향
<ol><li>0.3%, 0.5%, 0.8% 한천을 함유하는 반고체 배지를 준비하고, 상기와 같이 천자에 의해 접종한다. 배양 24-48 시간 후에 결과를 관찰하십시오.</li>
</ol>

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저자는 공개할 이해 상충이 없습니다.

반고체 배지 방법에 의한 박테리아 운동성의 검출은 많은 요인에 의해 영향을 받는다13,14. 산소(한천 표면의 호기성, 반고체 배지가 있는 튜브 바닥의 비호기성), pH 및 온도와 같은 박테리아 성장 조건은 박테리아 편모의 생존력에 영향을 미칠 수 있으며, 이는 운동성 감소 또는 운동성 상실로 이어질 수 있다15. 또한, 일부 점액형 박테?...

박테리아 운동성은 박테리아 병원성, 생물막 형성 및 약물 내성에 중요한 역할을 합니다1. 박테리아 운동성은 병원성과 밀접한 관련이 있으며, 숙주 세포의 초기 감염 동안 박테리아 집락화에 필요하다2. 생물막 형성은 박테리아 운동성과 밀접한 관련이 있으며, 박테리아는 운동성을 통해 고체 배지의 표면에 부착됩니다. 박테리아의 운동성은 오랫동안 생물막 형성과 양의 상관관계가 있는 것으로 여겨져 왔습니다. 생물막으로 인한 높은 수준의 박테리아 약물 내성은 인체 건강에 위협이되는 지속적인 감염을 유발할 수 있습니다 3,4,5. 따라서 박테리아의 운동성을 감지하는 것이 중요합니다. 박테리아 운동성 검사는 주로 살아있는 상태에서 다양한 형태의 박테리아의 운동성을 검사하는 데 사용되며, 이는 편모의 유무를 간접적으로 결정할 수 있으므로 박테리아 식별에 중요한 역할을 합니다.
박테리아의 운동성을 감지하는 직간접적인 방법이 있다6. 편모를 가진 박테리아는 운동성을 나타내기 때문에 편모의 유무를 감지하여 박테리아가 운동성인지 여부를 간접적으로 검출할 수 있습니다. 예를 들어, 전자 현미경 및 편모 염색에 의해 간접적으로 운동성을 감지하여 박테리아가 운동성이 있음을 나타낼 수 있습니다. 서스펜션 드롭 및 반고체 펑크 방법과 같은 직접적인 방법으로 감지하는 것도 가능합니다. 
세균 운동성을 감지하기 위해 학부 미생물학 실험실에서 일반적으로 사용되는 반고체 천자 방법은 세균 성장 방향에 따라 0.4-0.8% 한천을 함유한 반고체 한천 배지에 박테리아를 천자 내로 접종합니다. 박테리아가 펑크 라인을 따라 자라서 주변으로 퍼지면 구름 모양 (브러시 같은) 성장 흔적이 나타나 편모의 존재와 운동성을 나타냅니다. 펑크 라인 성장 흔적이 없으면 박테리아는 편모도 운동성도 없습니다. 
그러나이 방법은 박테리아가 무색이고 투명하며 편모 활성은 살아있는 박테리아 및 기타 요인의 생리적 특성과 한천의 농도 및 시험관의 작은 직경에 의해 영향을받는 단점이 있습니다. 또한, 호기성 박테리아는 한천 표면에서의 성장에만 적합하여 박테리아 운동성 관찰에 영향을 미칩니다. 따라서이 실험을 개선하기 위해 2,3,5- 트리 페닐 테트라 졸륨 클로라이드 (TTC) (무색)를 배지에 첨가하여 TTC 7,8,9,10의 적색 생성물 형성을 촉매하기 위해 세포 내 탈수소 효소를 사용하는 현재의 직접 천자 방법보다 박테리아 운동성을 결정하는 더 신뢰할 수 있고 직관적 인 방법을 확립했습니다.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Bacto Agar</td>
			<td>Difco</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Escherichia coli</td>
			<td>ATCC</td>
			<td>ATCC25922</td>
			<td>Positive control</td>
		</tr>
		<tr>
			<td>Pseudomonas aeruginosa</td>
			<td>ATCC</td>
			<td>ATCC27853</td>
			<td>Positive control</td>
		</tr>
		<tr>
			<td>Salmonella typhimurium</td>
			<td>ATCC</td>
			<td>ATCC14028</td>
			<td>Positive control</td>
		</tr>
		<tr>
			<td>Staphylococcus aureus</td>
			<td>ATCC</td>
			<td>ATCC25923</td>
			<td>Negative nonmotile control</td>
		</tr>
		<tr>
			<td>Tryptose</td>
			<td>&#160;OXOID</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>TTC</td>
			<td>Sigma</td>
			<td>298-96-4</td>
			<td></td>
		</tr>
		<tr>
			<td>VITEK 2 automated microbial identification system</td>
			<td>Bio M&#233;rieux</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

visualizing bacterial motility based on a color reaction

박테리아 운동성은 박테리아 병원성, 생물막 형성 및 약물 내성에 매우 중요합니다. 박테리아 운동성은 많은 병원성 종의 침입 및/또는 보급에 매우 중요합니다. 따라서 박테리아의 운동성을 감지하는 것이 중요합니다. 산소, pH 및 온도와 같은 박테리아 성장 조건은 박테리아 성장과 박테리아 편모의 발현에 영향을 줄 수 있습니다. 이것은 운동성을 감소시키거나 심지어 운동성을 상실하여 박테리아 운동성에 대한 부정확한 평가를 초래할 수 있습니다. 살아있는 박테리아의 세포 내 탈수소효소에 의한 2,3,5-트리페닐 테트라졸륨 클로라이드(TTC)의 색상 반응을 기반으로 박테리아 운동성 검출을 위해 전통적인 반고체 한천에 TTC를 첨가했습니다. 그 결과 박테리아 운동성 검출을 위한 이 TTC 반고체 한천 방법이 간단하고 작동하기 쉬우며 크고 값비싼 기구가 필요하지 않다는 것을 보여주었습니다. 그 결과, 0.3% 한천으로 제조된 반고체 배지에서 가장 높은 운동성이 관찰되는 것으로 나타났다. 기존의 반고체 매체와 비교하여 결과를 평가하기 쉽고 정확합니다.

운동성 검출을 위해 표준 균주와 분리된 균주 모두 비교하였으며, 그 결과를 표 1에 나타내었다. 편모가 없기 때문에 황색포도상구균과 폐렴간균은 전통적인 배지와 TTC 반고체 배지 모두에서 접종된 라인을 따라서만 성장했습니다. 대조적으로, 녹농균(Pseudomonas aeruginosa), 대장균(Escherichia coli) 및 살모넬라 티피뮤리움(Salmonella typhimurium)은 TTC 반고체 배?...

Watch this Scientific Journal Video about Visualizing Bacterial Motility Based on a Color Reaction at JoVE.com

Visualizing Bacterial Motility Based on a Color Reaction

여기에서는 색상 반응을 기반으로 박테리아 운동성을 감지하는 프로토콜을 제시합니다. 이 방법의 주요 장점은 평가하기 쉽고 정확하며 특수 장비가 필요하지 않다는 것입니다.

색상 반응을 기반으로 한 박테리아 운동성 시각화

Bacterial motility is crucial for bacterial pathogenicity, biofilm formation, and drug resistance. Bacterial motility is crucial for the invasion and/or ...

visualizing-bacterial-motility-based-on-a-color-reaction

Research

JoVE Journal

Biology

3.6K 조회수.  China Pharmaceutical University. 여기에서는 색상 반응을 기반으로 박테리아 운동성을 감지하는 프로토콜을 제시합니다. 이 방법의 주요 장점은 평가하기 쉽고 정확하며 특수 장비가 필요하지 않다는 것입니다.

비디오: Visualizing Bacterial Motility Based on a Color Reaction

Assessing Neural Stem Cell Motility Using an Agarose Gel-based Microfluidic Device

While microfluidic technology is reaching a new level of maturity for macromolecular assays, cell-based assays are still at an infant stage1. This is largely due to the difficulty with which one can create a cell-compatible and steady microenvironment using conventional microfabrication techniques and materials. We address this problem via the introduction of a novel microfabrication material, agarose gel, as the base material for the microfluidic device. Agarose gel is highly malleable, and permeable to gas and nutrients necessary for cell survival, and thus an ideal material for cell-based assays. We have shown previously that agarose gel based devices have been successful in studying bacterial and neutrophil cell migration2. In this report, three parallel microfluidic channels are patterned in an agarose gel membrane of about 1mm thickness. Constant flows with media/buffer are maintained in the two side channels using a peristaltic pump. Cells are maintained in the center channel for observation. Since the nutrients and chemicals in the side channels are constantly diffusing from the side to center channel, the chemical environment of the center channel is easily controlled via the flow along the side channels. Using this device, we demonstrate that the movement of neural stem cells can be monitored optically with ease under various chemical conditions, and the experimental results show that the over expression of epidermal growth factor receptors (EGFR) enhances the motility of neural stem cells. Motility of neural stem cells is an important biomarker for assessing cells aggressiveness, thus tumorigenic factor3. Deciphering the mechanism underlying NSC motility will yield insight into both disorders of neural development and into brain cancer stem cell invasion.

We demonstrate that the over expression of epidermal growth factor receptors (EGFR) enhances the motility of neural stem cells(NSCs) using a novel agarose gel based microfluidic device. This technology can be readily adaptable to other mammalian cell systems where cell sources are scarce, such as human neural stem cells, and the turn around time is critical.

아가로 오스 겔 기반 Microfluidic 장치를 사용 신경 줄기 세포의 운동성 평가

While microfluidic technology is reaching a new level of maturity for macromolecular assays, cell-based assays are still at an infant stage1. This is ...

연구

생물학

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA template is attached to a functionalized glass coverslip and replicated extensively after introduction of replication proteins and nucleotides (Figure 1). The growing product double-strand DNA (dsDNA) is extended with laminar flow and visualized by using an intercalating dye. Measuring the position of the growing DNA end in real time allows precise determination of replication rate (Figure 2). Furthermore, the length of completed DNA products reports on the processivity of replication. This experiment can be performed very easily and rapidly and requires only a fluorescence microscope with a reasonably sensitive camera.


This protocol demonstrates a simple single-molecule fluorescence microscopy technique for visualizing DNA replication by individual replisomes in real time.

형광 현미경과 함께 머릿속으로 단일 분자의 DNA 복제

We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA ...

Visualizing RNA Localization in Xenopus Oocytes

RNA localization is a conserved mechanism of establishing cell polarity. Vg1 mRNA localizes to the vegetal pole of Xenopus laevis oocytes and acts to spatially restrict gene expression of Vg1 protein. Tight control of Vg1 distribution in this manner is required for proper germ layer specification in the developing embryo. RNA sequence elements in the 3' UTR of the mRNA, the Vg1 localization element (VLE) are required and sufficient to direct transport. To study the recognition and transport of Vg1 mRNA in vivo, we have developed an imaging technique that allows extensive analysis of trans-factor directed transport mechanisms via a simple visual readout. 
To visualize RNA localization, we synthesize fluorescently labeled VLE RNA and microinject this transcript into individual oocytes. After oocyte culture to allow transport of the injected RNA, oocytes are fixed and dehydrated prior to imaging by confocal microscopy. Visualization of mRNA localization patterns provides a readout for monitoring the complete pathway of RNA transport and for identifying roles in directing RNA transport for cis-acting elements within the transcript and trans-acting factors that bind to the VLE (Lewis et al., 2008, Messitt et al., 2008). We have extended this technique through co-localization with additional RNAs and proteins (Gagnon and Mowry, 2009, Messitt et al., 2008), and in combination with disruption of motor proteins and the cytoskeleton (Messitt et al., 2008) to probe mechanisms underlying mRNA localization.

Visualizing RNA Localization in Xenopus Oocytes

Visualization of in vivo RNA transport is accomplished by microinjection of fluorescently labeled RNA transcripts into Xenopus oocytes, followed by confocal microscopy.

머릿속 RNA 현지화에서 Xenopus Oocytes

RNA localization is a conserved mechanism of establishing cell polarity.  Vg1 mRNA localizes to the vegetal pole of Xenopus laevis oocytes and acts to ...

Gastrointestinal Motility Monitor (GIMM)

The Gastrointestinal Motility Monitor (GIMM; Catamount Research and Development; St. Albans, VT) is an in vitro system that monitors propulsive motility in isolated segments of guinea pig distal colon. The complete system consists of a computer, video camera, illuminated organ bath, peristaltic and heated water bath circulating pumps, and custom GIMM software to record and analyze data. Compared with traditional methods of monitoring colonic peristalsis, the GIMM system allows for continuous, quantitative evaluation of motility. The guinea pig distal colon is bathed in warmed, oxygenated Krebs solution, and fecal pellets inserted in the oral end are propelled along the segment of colon at a rate of about 2 mm/sec. Movies of the fecal pellet proceeding along the segment are captured, and the GIMM software can be used track the progress of the fecal pellet. Rates of propulsive motility can be obtained for the entire segment or for any particular region of interest. In addition to analysis of bolus-induced motility patterns, spatiotemporal maps can be constructed from captured video segments to assess spontaneous motor activity patterns. Applications of this system include pharmacological evaluation of the effects of receptor agonists and antagonists on propulsive motility, as well as assessment of changes that result from pathophysiological conditions, such as inflammation or stress. The guinea pig distal colon propulsive motility assay, using the GIMM system, is straightforward and simple to learn, and it provides a reliable and reproducible method of assessing propulsive motility.

Gastrointestinal Motility Monitor GIMM

Evaluation of colonic motility in the guinea pig distal colon with the Gastrointestinal Motility Monitor (GIMM) is a straightforward and simple to learn approach to quantitatively evaluate propulsive motility in the gastrointestinal tract.

The Gastrointestinal Motility Monitor (GIMM; Catamount Research and Development; St. Albans, VT) is an in vitro system that monitors propulsive motility ...

Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays

Bacterial surface motility, such as swarming, is commonly examined in the laboratory using plate assays that necessitate specific concentrations of agar and sometimes inclusion of specific nutrients in the growth medium. The preparation of such explicit media and surface growth conditions serves to provide the favorable conditions that allow not just bacterial growth but coordinated motility of bacteria over these surfaces within thin liquid films. Reproducibility of swarm plate and other surface motility plate assays can be a major challenge. Especially for more &#8220;temperate swarmers&#8221; that exhibit motility only within agar ranges of 0.4%-0.8% (wt/vol), minor changes in protocol or laboratory environment can greatly influence swarm assay results. &#8220;Wettability&#8221;, or water content at the liquid-solid-air interface of these plate assays, is often a key variable to be controlled. An additional challenge in assessing swarming is how to quantify observed differences between any two (or more) experiments. Here we detail a versatile two-phase protocol to prepare and image swarm assays. We include guidelines to circumvent the challenges commonly associated with swarm assay media preparation and quantification of data from these assays. We specifically demonstrate our method using bacteria that express fluorescent or bioluminescent genetic reporters like green fluorescent protein (GFP), luciferase (lux operon), or cellular stains to enable time-lapse optical imaging. We further demonstrate the ability of our method to track competing swarming species in the same experiment.

Swarming motility is influenced by physical and environmental factors. We describe a two-phase protocol and guidelines to circumvent the challenges commonly associated with swarm assay preparation and data collection. A macroscopic imaging technique is employed to obtain detailed information on swarm behavior that is not provided by current analysis techniques.

세균 표면 운동성 분석의 준비, 이미징 및 정량화

Bacterial surface motility, such as swarming, is commonly examined in the laboratory using plate assays that necessitate specific concentrations of agar ...

Monitoring Endoplasmic Reticulum Calcium Homeostasis Using a Gaussia Luciferase SERCaMP

The endoplasmic reticulum (ER) contains the highest level of intracellular calcium, with concentrations approximately 5,000-fold greater than cytoplasmic levels. Tight control over ER calcium is imperative for protein folding, modification and trafficking. Perturbations to ER calcium can result in the activation of the unfolded protein response, a three-prong ER stress response mechanism, and contribute to pathogenesis in a variety of diseases. The ability to monitor ER calcium alterations during disease onset and progression is important in principle, yet challenging in practice. Currently available methods for monitoring ER calcium, such as calcium-dependent fluorescent dyes and proteins, have provided insight into ER calcium dynamics in cells, however these tools are not well suited for in vivo studies. Our lab has demonstrated that a modification to the carboxy-terminus of Gaussia luciferase confers secretion of the reporter in response to ER calcium depletion. The methods for using a luciferase based, secreted ER calcium monitoring protein (SERCaMP) for in vitro and in vivo applications are described herein. This video highlights hepatic injections, pharmacological manipulation of GLuc-SERCaMP, blood collection and processing, and assay parameters for longitudinal monitoring of ER calcium.

Monitoring Endoplasmic Reticulum Calcium Homeostasis Using a Gaussia Luciferase SERCaMP

Endoplasmic reticulum calcium homeostasis is disrupted in diverse pathologies. A secreted ER calcium monitoring protein (SERCaMP) reporter can be used to detect disruptions in the ER calcium store. This protocol describes the use of a Gaussia luciferase SERCaMP to examine ER calcium homeostasis in vitro and in vivo.

사용 소포체 칼슘 항상성 모니터링<em&gt; Gaussia</em&gt; 루시 페라 제 SERCaMP

The endoplasmic reticulum (ER) contains the highest level of intracellular calcium, with concentrations approximately 5,000-fold greater than cytoplasmic ...

Measurement and Analysis of Extracellular Acid Production to Determine Glycolytic Rate

Extracellular measurement of oxygen consumption and acid production is a simple and powerful way to monitor rates of respiration and glycolysis1. Both mitochondrial (respiration) and non-mitochondrial (other redox) reactions consume oxygen, but these reactions can be easily distinguished by chemical inhibition of mitochondrial respiration. However, while mitochondrial oxygen consumption is an unambiguous and direct measurement of respiration rate2, the same is not true for extracellular acid production and its relationship to glycolytic rate 3-6. Extracellular acid produced by cells is derived from both lactate, produced by anaerobic glycolysis, and CO2, produced in the citric acid cycle during respiration. For glycolysis, the conversion of glucose to lactate- + H+ and the export of products into the assay medium is the source of glycolytic acidification. For respiration, the export of CO2, hydration to H2CO3 and dissociation to HCO3- + H+ is the source of respiratory acidification. The proportions of glycolytic and respiratory acidification depend on the experimental conditions, including cell type and substrate(s) provided, and can range from nearly 100% glycolytic acidification to nearly 100% respiratory acidification 6. Here, we demonstrate the data collection and calculation methods needed to determine respiratory and glycolytic contributions to total extracellular acidification by whole cells in culture using C2C12 myoblast cells as a model.

Glycolysis is a defining metabolic marker in multiple biological systems. Monitoring glycolysis by measuring the extracellular flux of H+ is common, but requires correction to be quantitative and unambiguous. Here, we demonstrate how to gather and correct extracellular flux data to distinguish between respiratory and glycolytic sources of extracellular acidification.

세포 외 산 생산의 측정 및 분석은 당분 속도를 확인하는

Extracellular measurement of oxygen consumption and acid production is a simple and powerful way to monitor rates of respiration and glycolysis1. Both ...

SA-&#946;-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs

Cell senescence is one of the hallmarks of aging known to negatively influence a healthy lifespan. Drugs able to kill senescent cells specifically in cell culture, termed senolytics, can reduce the senescent cell burden in vivo and extend healthspan. Multiple classes of senolytics have been identified to date including HSP90 inhibitors, Bcl-2 family inhibitors, piperlongumine, a FOXO4 inhibitory peptide and the combination of Dasatinib/Quercetin. Detection of SA-&#946;-Gal at an increased lysosomal pH is one of the best characterized markers for the detection of senescent cells. Live cell measurements of senescence-associated &#946;-galactosidase (SA-&#946;-Gal) activity using the fluorescent substrate C12FDG in combination with the determination of the total cell number using a DNA intercalating Hoechst dye opens the possibility to screen for senotherapeutic drugs that either reduce overall SA-&#946;-Gal activity by killing of senescent cells (senolytics) or by suppressing SA-&#946;-Gal and other phenotypes of senescent cells (senomorphics). Use of a high content fluorescent image acquisition and analysis platform allows for the rapid, high throughput screening of drug libraries for effects on SA-&#946;-Gal, cell morphology and cell number.

SA-&amp;#946;-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs

Cellular senescence is the key factor in the development of chronic age-related pathologies. Identification of therapeutics that target senescent cells show promise for extending healthy aging. Here, we present a novel assay to screen for the identification of senotherapeutics based on measurement of senescence associated &#946;-Galactosidase activity in single cells.

SA-β-갈락토시다아제 계 선별 분석법

Cell senescence is one of the hallmarks of aging known to negatively influence a healthy lifespan. Drugs able to kill senescent cells specifically in cell ...

Ex Vivo Method for Assessing the Mouse Reproductive Tract Spontaneous Motility and a MATLAB-based Uterus Motion Tracking Algorithm for Data Analysis

Dysmenorrhea, or painful cramping, is the most common symptom associated with menses in females and its severity can hinder women&#39;s everyday lives. Here, we present an easy and inexpensive method that would be instrumental for testing new drugs decreasing uterine contractility. This method utilizes the unique ability of the entire mouse reproductive tract to exhibit spontaneous motility when maintained ex vivo in a Petri dish containing oxygenated Krebs buffer. This spontaneous motility resembles the wave-like myometrial activity of the human uterus, referred to as endometrial waves. To demonstrate the effectiveness of the method, we employed a well-known uterine relaxant drug, epinephrine. We demonstrate that the spontaneous motility of the entire mouse reproductive tract can be quickly and reversibly inhibited by 1 &#181;M epinephrine in this Petri dish model. Documenting the changes of uterine motility can be easily done using an ordinary smart phone or a sophisticated digital camera. We developed a MATLAB-based algorithm allowing motion tracking to quantify spontaneous uterine motility changes by measuring the rate of uterine horn movements. A major advantage of this ex vivo approach is that the reproductive tract remains intact throughout the entire experiment, preserving all intrinsic intrauterine cellular interactions. The major limitation of this approach is that up to 10-20% of uteri may exhibit no spontaneous motility. Thus far, this is the first quantitative ex vivo method for assessing spontaneous uterine motility in a Petri dish model.

Uterine contractions are important for the well-being of females. However, pathologically increased contractility may result in dysmenorrhea, especially in younger females. Here, we describe a simple ex vivo preparation allowing quick assessment of the efficacy of smooth muscle relaxants that may be used for treating dysmenorrhea.

마우스 생식 기관 자발적 운동성 및 데이터 분석을 위한 MATLAB 기반 자궁 운동 추적 알고리즘을 평가하기 위한 Ex Vivo 방법

Dysmenorrhea, or painful cramping, is the most common symptom associated with menses in females and its severity can hinder women&#39;s everyday lives. ...

A Capsule-Based Model for Immature Hard Tick Stages Infestation on Laboratory Mice

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of mouse models in tick research is critical for understanding their biology and tick-host-pathogen interactions. Here we demonstrate a non-laborious technique for the feeding of immature stages of hard ticks on laboratory mice. The benefit of the method is its simplicity, short duration, and the ability to monitor or collect ticks at different time points of an experiment. In addition, the technique allows attachment of two individual capsules on the same mouse, which is beneficial for a variety of experiments where two different groups of ticks are required to feed on the same animal. The non-irritating and flexible capsule is made from easily accessible materials and minimizes the discomfort of the experimental animals. Furthermore, euthanasia is not necessary, mice recover completely after the experiment and are available for re-use.

In this study, a feeding system for nymphal and larval stages of hard tick was developed using a capsule attached to laboratory mouse. The feeding capsule is made from flexible materials and remains firmly attached to the mouse for at least one week and allows comfortable monitoring of tick feeding.

실험실 마우스에 미숙한 하드 진드기 단계 감염에 대 한 캡슐 기반 모델

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of ...