Name: nanosensors to detect protease activity in vivo for noninvasive diagnostics
Uploaded: 2018-07-16T14:30:00
Description: Watch this Scientific Journal Video about Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics at JoVE.com

이 작품은 NIH 감독의 새로운 혁신 상 수상 (제에 의해 투자 되었다 DP2HD091793)입니다. NSF 대학원 연구 장학 프로그램 (보조금 번호 Q.D.M.는 지원 DGE-1650044). B.A.H는 국립 연구소의 건강 GT BioMAT 훈련 그랜트 보너스 번호 5T32EB006343에서 조오지 아 기술 대통령의 친교에 의해 지원 됩니다. G.A.K.는 버로우즈 환영 기금에서 과학적인 인터페이스에서 경력 수상을 보유 하고있다. 내용은 전적으로 저자의 책임 이며 반드시 국립 보건원의 공식 의견을 대표 하지 않는다.

연구원의 교육 기관에서 기관 동물 관리 및 사용 위원회 (IACUC)에서 기관 승인 다음 동물 실험을 수행 하는 데 필요한입니다. 또한, 표준 동물 보호 시설 (예를 들어, 챔버, 무 균 동물 후드, anesthetization, isofluorane 챔버 및 윤리적인 끝점 euthanization에 대 한 공동2 챔버)는이 제대로 수행 하는 데 필요한 실험입니다. 특별 한 교육 및 이러한 시설 지원 하나의 기관에 생리 연구 실험실 (PRL) ...

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이 방법은 나노 코어를 활용 하는 프로 테아 제 기판으로 구성 된 활동 기반 nanosensors의 개발을 설명 합니다. 쪼개진된 펩 티 드 제품은 5 nm23 및 비 침 투 적인 신호를 생성 하는 소변으로 필터의 신장 크기 여과 한계 보다 작은 때문에 분해 분열의 이벤트 "pharmacokinetic 스위치", 불리는. 따라서, 그것은 유체역학 반경 5 보다 큰 나노 입자 또는 운반대를 사용 하는 것이 중요, 아무것?...

프로 테아 제 펩 티 드 결합의 가수분해를 전문화 하 고 항상성, 질병1, allostasis, 등 많은 생물 학적 과정을 중요 한 통제가 다기능 효소는. 프로 테아 제 활동의 변경된 상태 질병, 암 및 심장 혈관 질병, 프로 테아 제 임상 생체2,3으로 개발을 위한 매력적인 후보자를 만드는 등의 다양 한 상관 되었다. 또한, 프로 테아 제 활동은 별개에 기능적으로 연결 된 pathogeneses, 환자 결과 및4질병 예 후. 넓게, 바이오 센서 개발 되었습니다 다양 한 생물 학적 현상을 감지 하 고 질병, 암, 신경 질환, 전자 등 처리5,6,,78 , 9. 좀 더 구체적으로, 기판 기반 효소 센서 프로 테아 제 활동을 감지 하 여 진단 이미징10 fluorogenic 프로브 포함 개발 되었고 isotopically 펩 티 드 기질에 시험관 에 대 한 표시 질량 분석11탐지입니다. 또한, 활동 기반 프로브 개발 되었습니다, 바인딩하거나 대상 프로 테아 제12수정 기판 같은 영역을 포함 하는. 이 방법으로 활성 사이트 수정 하며 분석 조직, vivo에서 응용 프로그램을 제한의 수확 때 대상 효소 저해 irreversibly는. 그러나, 효소 활동의 규칙은 내 생 억제제의 존재와 같은 다른 생물 학적 활동의 문맥에 크게 의존 하기 때문에 효소 활동을 vivo에서, 감각을 중요 하다.
이 작품의 목표는 소변에서 측정 신호를 생산 하 여 효소 활동에서 vivo에서 검색 활동 기반 nanosensors의 공식 설명. 이 플랫폼 기능 biomarker로 dysregulated 프로 테아 제 활동을 사용 하 여 암과 같은 복잡 한 질병을 판별 하는 비 침범 성 진단으로 사용 됩니다. 산화 철 나노 입자 (IONP) 효소 기질에 활용 된 nanosensor 플랫폼에 의하여 이루어져 있다. 이러한 기판 프로 테아 제 쪼개 기판 때 출시 되는 형광 기자에 의해 종료 됩니다. 이러한 IONPs vivo에서순환, 질병 사이트, 지역화 및 기판 활성 질병 관련 된 프로 테아 제에 노출. 분열, 후 형광 기자 출시 되 고, 그들의 작은 크기로 인해 필터링 됩니다 소변에는 IONP에 uncleaved 기판 시체에 남아 있는 동안. 따라서, 효소 활동에서 vivo에서 증가 소변 (그림 1)에서 리포터의 높은 농도 발생 합니다. 우리의 플랫폼 소변 검사 이기 때문에, 아무 이미징 플랫폼 필수 이며 진단 신호는 소변에서 풍성 하 게.
이 플랫폼은 다양 한 질병 암, 섬유 증, 혈전 증13,14등을 감지 하 설계 될 수 있다. 여기 우리가 설명 하는 매트릭스 metallopeptidase 9 (MMP9)에서 고도 검색 하기 위해 nanosensors의 디자인 활동으로 대 장 암의 바이오 마커. 대 장 암은 약된 136,800 새로운 사례와 2014 혼자1550,300 죽음, 미국에서 암 사망의 두 번째 주요 원인이. 대 장 종양 세포 생성 MMP9 전이16뿐만 아니라 악성 진행, 매트릭스 저하, 운전 표시 되었습니다. 또한, 우리 문학17에서 MMP9에 대 한 적합 한 펩 티 드 기질 (PLGVRGK) 확인. 조기 암 발견 및 저가 포인트의 케어 진단13,,1418,19,20,21이 플랫폼을 사용할 수 있습니다.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57937/57937fig1.jpg"> 그림 1: Nanosensor 활동 비보의 도식. Nanosensors는 본문을 통해 순환 하 고 질병의 사이트 지역화. 다음, 질병 관련 된 프로 테아 제 펩 티 드 기질 IONPs를 제시한 다니엘. 쪼개진된 파편의 크기 신장 클리어런스, 그들이 소변에서 지역화 하는 원인에 대 한 수 있습니다. 동물 소변 후 그들의 기자 분자에 의해이 펩 티 드 파편을 분석할 수 있습니다. <a href="https://www.jove.com/files/ftp_upload/57937/57937fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭 하십시오.</a>

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			<td height="21" style="height:21px;width:375px;">0.2 &#181;m syringe filters</td>
			<td style="width:171px;">VWR</td>
			<td style="width:233px;">4652</td>
			<td style="width:167px;"></td>
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			<td height="21" style="height:21px;width:375px;">18G needle</td>
			<td style="width:171px;">VWR</td>
			<td style="width:233px;">89134-024</td>
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			<td height="21" style="height:21px;width:375px;">15 mL conicals</td>
			<td style="width:171px;">VWR</td>
			<td style="width:233px;">89039-670</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:375px;">250 mL Erlenmeyer flask</td>
			<td style="width:171px;">VWR</td>
			<td style="width:233px;">89000-362</td>
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			<td height="21" style="height:21px;width:375px;">Stir bar</td>
			<td style="width:171px;">VWR</td>
			<td style="width:233px;">58949-006</td>
			<td></td>
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			<td height="21" style="height:21px;width:375px;">Hot Plate/Magnetic Stirrer</td>
			<td style="width:171px;">VWR</td>
			<td style="width:233px;">97042-634</td>
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			<td height="21" style="height:21px;width:375px;">Glacial acetic acid</td>
			<td style="width:171px;">VWR</td>
			<td style="width:233px;">97064-482</td>
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			<td height="21" style="height:21px;width:375px;">Albumin from Bovine Serum (BSA)</td>
			<td style="width:171px;">Thermo Fisher</td>
			<td style="width:233px;">A13100</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Iron (III) chloride hexahydrate</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">236489</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Iron (II) chloride tetrahydrate</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">44939</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Epichlorohydrin</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">45340-500ML-F</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">DMF</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">D4551</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Ammonium Hydroxide</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">320145-500ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Sodium Hydroxide pellets</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">221465-500G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">EDTA</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">E9884</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Sodium Borate</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">B9876</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">L-Cysteine</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">168149-100G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Tris-HCl</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">T5941</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Tris base</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">T6066</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">PBS tablets</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:233px;">P4417</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Dextran</td>
			<td style="width:171px;">Pharmacosmos</td>
			<td>5510 0020 9006</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Amicon 15 mL 10k filters, 24 pk</td>
			<td style="width:171px;">Millipore</td>
			<td style="width:233px;">UFC901024</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Amicon 15 mL 30k filters, 24 pk</td>
			<td style="width:171px;">Millipore</td>
			<td style="width:233px;">UFC903024</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Amicon 15 mL 100k filters, 24 pk</td>
			<td style="width:171px;">Millipore</td>
			<td style="width:233px;">UFC910024</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Zetasizer Nano ZS</td>
			<td style="width:171px;">Malvern Panalytical</td>
			<td style="width:233px;">NanoZS</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Slide-A-Lyzer Dialysis Cassette</td>
			<td style="width:171px;">LifeTech</td>
			<td style="width:233px;">66130</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Dynabeads MyOne Tosylactivated</td>
			<td style="width:171px;">LifeTech</td>
			<td style="width:233px;">65501</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">SIA</td>
			<td style="width:171px;">Life Tech</td>
			<td style="width:233px;">22349</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">PEG 20k</td>
			<td style="width:171px;">Laysan Bio</td>
			<td style="width:233px;">MPEG-SH-20K-1g</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Fluorescein Antibody [2A3]</td>
			<td style="width:171px;">GeneTex</td>
			<td style="width:233px;">GTX10257</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Hiload 16/600 superdex 200</td>
			<td style="width:171px;">GE Healthcare</td>
			<td style="width:233px;">45-002-490</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">Plate Reader</td>
			<td style="width:171px;">Fisher</td>
			<td style="width:233px;">BTCYT5M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:375px;">BD Insulin Syringes</td>
			<td style="width:171px;">Fisher</td>
			<td style="width:233px;">NC0872854</td>
			<td></td>
		</tr>
	</tbody>
</table>

nanosensors to detect protease activity in vivo for noninvasive diagnostics

프로 테아 제는 다기능 효소 펩 티 드 결합의 가수분해에서 전문 및 항상성 등 allostasis 광범위 한 생물 학적 과정을 제어. 또한, dysregulated 프로 테아 제 활동 pathogenesis 드라이브 이며 암;과 같은 질병의 기능 바이오 마커 따라서, 효소 활동에서 vivo에서 감지 하는 능력은 생물 의학 진단에 대 한 임상 관련 정보를 제공할 수 있습니다. 이 프로토콜의 목표 조사 효소 활동에서 vivo에서 소변에서 정량 신호를 생산 하 여 nanosensors를 만드는 것입니다. 이러한 효소 nanosensors의 두 가지 구성 요소 구성: 나노 및 기판. 대상 질병 사이트에 순환 반감기 및 기판 납품 증가를 나노 기능. 기판은 짧은 펩 티 드 순서 (6-8 AA), 대상 효소 또는 프로 테아의 그룹에 구체적으로 설계 된입니다. 기판은 나노 입자의 표면에 활용 하 고 탐지를 위한 형광 표식 같은 기자에 의해 종료 됩니다. Dysregulated 프로 테아 제 펩 티 드 기질 다니엘로 기자 프로 테아 제 활동의 한 biomarker로 정량화에 대 한 소변으로 필터링 됩니다. 여기는 매트릭스 metalloproteinase 9 (MMP9), 종양 진행 및 전이, 대 장 암 마우스 모델에서의 검출에 대 한 관련 된에 대 한 nanosensor의 건설에 설명 합니다.

대부분의 인구는 IONPs의 평균 직경 40-50 nm에서 범위는 주위에 이다. Pegylation, 후이 크기 범위는 약 6 시간13 생체 내에서 의 순환 반감기 ( 그림 2a참조). 특정 크기 범위에 대 한 선택 하 고 싶다면 하나 하나 다른 직경을 가진 IONP 분수 분리 하 크기 배제 크로마토그래피를 사용할 수 있습니다. 가장 하 여 나노 입자 dextran의 외부...

Watch this Scientific Journal Video about Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics at JoVE.com

Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics

프로 테아 제는 기본적인 생물학 과정 및 암과 같은 복잡 한 질병의 dysregulated 프로 테아 제 활동 드라이브 진행에 관련 된 단단히 규제 효소. 이 방법의이 목표 호스트 소변에서 감지 하 고 질병을 차별 분열 신호를 생산 하 여 효소 활동에서 vivo에서 측정 하는 nanosensors를 만드는 것입니다.

Nanosensors 프로 테아 제 활동에 감지 비보에 비 침 투 적인 진단에 대 한

Proteases are multi-functional enzymes that specialize in the hydrolysis of peptide-bonds and control broad biological processes including homeostasis and ...

This method can help answer key questions in protea mix and diagnostics, such as how to design probes that can detect the progression of diseases by measuring proteates'activity in vivo. The main advantage of this method is that the diagnositics developed will be noninvasive, and can actually amplify the detection of biological signals. This method can be applied to numerous diseases, because many proteuses are actually major drivers of pathogenesis.In the chemical fume hood, transfer one ml of 30%ammonium hydroxide to a 15 ml conical tube. Place this tube into an ice bucket and let it chill for 30 minutes. Wash a 250 ml Ehrlenmeyer flask with deionized water.Then add 10 ml of double distilled water. Submerge the flask in an ice bath resting on a heat plate. Using a plastic pipette, begin the flow of nitrogen gas into the double distilled water in the flask.Bubble for 15 minutes to deoxygenate. Next, weigh out 4.5 g of dextrine in a 15 ml conical tube. Add water such that the final volume of the mixture including the dextrine is 20 ml.Vortex vigorously to dissolve the dextrine. Weigh out 78.5 mg of iron 3 chloride hexahydrate and add this to the dextrine solution. Vortex to dissolve.Using a 0.2 um filter, filter the resulting solution. After this, transfer 9 ml of the deoxygenated water to the 15 ml conical tube containing the 1 ml of chilled ammonium hydroxide. Return the conical tube to the ice bucket to chill.Weigh out 73 mg of iron 2 chloride tetrahydrate and dissolve it in the remaining 1 ml of deoxygenated double distilled water. Use a 0.2 um filter to filter the solution. Then, transfer the dextrine iron 3 solution into the 250 ml Ehrlenmeyer flask.Deoxygenate with nitrogen gas for 15 minutes on ice, while mixing at 1, 600 rpm. Cap the flask with a rubber septum. Using an 18 gauge needle, puncture the septum to flow nitrogen gas into the flask.Insert a separate 18 gauge needle, which will be the flow outlet. Next, use a 1 ml syringe to add 467 ul of the iron 2 solution to the dextrine iron 3 solution. Stir at 1, 600 rpm.Add the chilled dilute ammonium hydroxide solution dropwise to initiate the nucleation process. Make sure each drop mixes well before adding another. Stop the flow of nitrogen gas and remove the rubber septum.Remove the ice bath and replace it with a bath of warm water. Heat the solution to 75 degrees Celsius and then incubate for 75 minutes. Remove the flask from the hot plate.Doubly filter the solution through a 0.2 um filter and then a 0.1 um filter to remove any coarse particles. Using 100 kDa molecular weight cutoff concentrators, buffer exchange the particles into double distilled water to remove excess dextrine. If the flow through remains dark after two to three spins, replace the filters as they may have broken.Next, resuspend the iron oxide nanoparticles in double distilled water at a concentration of 10 mg per ml. Add 1.6 volumes of 5 molar sodium hydroxide, then add 0.65 volumes of epichlorohydrine. Using a plate shaker, rigorously mix at room temperature for 12 hours.Use a 20 ml syringe with an 18 gauge needle to transfer the iron oxide nanoparticle solution into a dialysis membrane with a 50 kDa molecular weight cutoff. Place the dialysis membrane into 4 liters of double distilled water. Incubate at room temperature overnight.After determining the concentration, bring the iron oxide nanoparticles to a concentration of 5 mg per ml. Add ammonium hydroxide to reach 20%and shake at room temperature for at least 12 hours to amenate the surface of the iron oxide nanoparticles. Then, buffer exchange using 30 kDa molecular weight cutoff filters.Use dynamic light scattering to determine the hydrodynamic radius of the iron oxide nanoparticles as outlined in the text protocol. After synthesizing a peptide substrate, aliquot 0.5 mg of iron oxide nanoparticles into the 10 kDa molecular weight cutoff spin filter tube. Buffer exchange into coupling buffer using a 10 kDa molecular weight cutoff spin filter as outlined in the text protocol.Dissolve SIA in DMF to reach a concentration of approximately 30 mg per ml. Then, add SIA to the iron oxide nanoparticles at a mole ratio of 500 to one. After incubation and buffer exchange, bring the final product to a concentration of 1 mg per ml.Mix the peptide of interest with 20 kDa thiol terminated polyethylene glycol, and then mix this peptide glycol solution with iron oxide nanoparticles. Cover the tubes in foil to prevent photo bleaching the fluorescent molecules. And incubate overnight at room temperature on a plate shaker set the to the highest speed.Next, add L cystine at a molar ratio of 500 to one to the iron oxide nanoparticles. Incubate for one hour at room temperature on a shaker set to the highest speed. After this, make an 18 ul nanoparticle solution using PVS, and a 200 nanomolar concentration of peptide.Add 2 ul of the protease of interest. Incubate in a plate reader for one hour at 37 degrees Celsius taking flourescence measurements every one to two minutes to monitor cleavage. Immediately after injecting the prepared nanosensor solution via tail vein injection, place the mice into metabolic cages, and note the time of injection for each mouse.Combine 2 ul of urine with 5 ul of prepared magnetic beads. Using PVS, bring the final total volume to 50 ul. Incubate at room temperature for 60 minutes.Next, wash twice using 50 ul of PVS each time, while using a magnetic separator to collect the magnetic beads after each wash. Elute twice with 32.5 ul of 5%glacial acetic acid. Use 35 ul of 2 molar tris to neutralize the pool dilution and achieve a final ph around 7.After this, use a plate reader at appropriate excitation and emission wavelengths to quantify the urine flourescence. Calculate the concentration of fluorescent reporter against a ladder of known concentrations of free flourophor. In this study, activity based nanosensors are developed by conjugating protease substrates to a nanoparticle core.The average diameter of iron oxide nanoparticles is observed to be between 40 and 50 nm. After pecolation, the circulation half life for this size range is seen to be approximately 6 hours. After successful peptide conjugation, spectral absorbance analysis reveals a distinct absorbance peak at the maximum excitation wavelength of the fluorescent reporter.The ability of the probe to sense protease activity is then tested by incubating an aliquot of nanosensors with recombinant proteases. It is typical to observe substrate cleavage velocities that follow Michaelis-Menten kinetics. Nanosensors coated with a near infrared fluorescent reporter are then used to visualize the pharmacokinetics in mouse models of cancer by whole animal fluorescent imaging.A fluorescent signal is seen to localize in the urine of mice, bearing LS 174 t colorectal, 60 to 90 minutes after intravenous administration of the nanosensors. Significantly lower fluorescent signals are observed in the bladders of control mice. It should be noted that the signal observed in the liver arises because particles are eventually scavenged by monocites and and macrophages in the ridiculoendophilial system, which are commonly found in the liver, spleen, and lymph nodes.Once mastered, this technique can be done in four days, if it's performed properly. While attempting this procedure, it's important to remember that flourescence is the primary readout, so it's important to keep the peptide substrates with fluorescent markers in the dark. Following this procedure, other methods like biodistribution studies and in viva imaging can be performed in order to answer additional questions, like localization of particles to various organs.After its development, this technique paved the way for researchers in the field of diagnostics to explore cancer biomarkers in mice. After watching this video, you should have a good understanding of how to synthesize and characterize iron oxide nanoparticles, use these to build nanosensors, then use the nanosensors in vivo. Don't forget that working with the chemicals involved in a nanoparticle synthesis can be extremely hazardous, and precautions such as personal protective equipment and working within a fume hood should always be taken while performing this procedure.

Research

JoVE Journal

Bioengineering

Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

의 순환 압력 생물 반응기 설계 예 VIVO</em대동맥 심장 밸브> 학습

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic ...

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Quantitative FRET (F&#246;rster Resonance Energy Transfer) Analysis for SENP1 Protease Kinetics Determination

Quantitative FRET F&#246;rster Resonance Energy Transfer Analysis for SENP1 Protease Kinetics Determination

정량은 SENP1 단백 분해 효소 반응 속도론 결정에 대해 (포스터 공명 에너지 전송) 분석 안달

Reversible posttranslational modifications of proteins with ubiquitin or ubiquitin-like proteins (Ubls) are widely used to dynamically regulate protein ...

FIBS-enabled Noninvasive Metabolic Profiling

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In the era of computational biology, new high throughput experimental systems are necessary in order to populate and refine models so that they can be ...

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

전체 피부 결함 모델은 생체의 혈관 신생을 평가하기<em&gt; 생체</em

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to ...

Magnetic Levitation Coupled with Portable Imaging and Analysis for Disease Diagnostics

질병 진단을위한 휴대용 이미징 및 분석과 결합 자기 부상

Currently, many clinical diagnostic procedures are complex, costly, inefficient, and inaccessible to a large population in the world. The requirements for ...

Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen

Foodborne 병원 체를 검출 하는 전기 화학 DNA 바이오 센서의 개발

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, ...

Calorespirometry: A Powerful, Noninvasive Approach to Investigate Cellular Energy Metabolism

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Many cell lines used in basic biological and biomedical research maintain energy homeostasis through a combination of both aerobic and anaerobic ...

Rigid Embedding of Fixed and Stained, Whole, Millimeter-Scale Specimens for Section-free 3D Histology by Micro-Computed Tomography

엄밀한 포함의 고정 및 스테인드, 전체, 마이크로 단층에 의해 섹션-무료 3D 조직학에 대 한 밀리미터 단위 표본

For over a hundred years, the histological study of tissues has been the gold standard for medical diagnosis because histology allows all cell types in ...

Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics

혈액학적 진단을 위한 폴리머 물방울 발생기 장치를 사용하여 메틸화 특정 멀티플렉스 물방울 PCR

A multiplexed droplet PCR (mdPCR) workflow and detailed protocol for determining epigenetic-based white blood cell (WBC) differential count is described, ...

Developing 3D Organized Human Cardiac Tissue within a Microfluidic Platform

미세 유체 플랫폼 내에서 3D 조직 된 인간 심장 조직 개발

The leading cause of death worldwide persists as cardiovascular disease (CVD). However, modeling the physiological and biological complexity of the heart ...