Description
There are five fluorescent control liposome products (Fluoroliposome®) for Clodrosome® (clodronate liposomes). All five fluorescent liposomes incorporate a lipophilic dye inside their membranes. They are insoluble in water; however, their fluorescence is easily detected when incorporated into membranes. DiI, DiO, DiD, DiR and DiA cover a wide range of excitation and emission wavelengths from 300s to 900s. DiI and DiO have fluorescence excitation and emission maxima separated by about 65 nm, facilitating two-color labeling. The emission spectrum of DiA is very broad, allowing it to be detected as green, orange or even red fluorescence depending on the optical filter used. DiI, DiO, DiD and DiR belong to the dialkylcarbocyanines family of compounds. The spectral properties of the dialkylcarbocyanines are largely independent of the lengths of the alkyl chains. Instead, they are determined by the heteroatoms in the terminal ring systems and the length of the connecting bridge. They have extremely high extinction coefficients, moderate fluorescence quantum yields and short excited state lifetimes in lipid environments (~1 ns). The fluorescence spectrum of each dye is shown below.
You can choose the Fluoroliposome® based on the type of the fluorescent equipment and filters that you use in your lab. Clodronate liposomes cannot be made fluorescent simply due to the potential for inaccurate and/or uninterpretable data being generated by labelled Clodrosome®. For more information, please refer to the technical note section.
Technical Information
Fluoroliposome®-DiO
Lipid Composition | Concentration (mg/ml) | Concentration (mM) | Molar Ratio Percentage |
---|---|---|---|
Total | 23 mg/ml | 35.1 mM | 100 |
L-alpha-Phosphatidylcholine | 18.8 | 24.3 | 70 |
Cholesterol | 4.2 | 10.9 | 30 |
Fluorescent Dye | Excitation/Emission (nm) | Concentration (mg/ml) | Concentration (mM) |
---|---|---|---|
3,3'-Dilinoleyloxacarbocyanine Perchlorate (DiO) | 484/501 | 0.0625 | 0.071 |
Buffer and Liposome Size | Specification |
---|---|
Buffer | Phosphate Buffered Saline |
pH | 7.4 |
Liposome Size | 1.5-2 µm |
Technical Notes
- The issue with fluorescent Clodrosome® has to do with the potential for inaccurate and/or uninterpretable data being generated by labelled Clodrosome®. When Clodrosome® induces macrophage apoptosis, the fluorescent lipid incorporated into the Clodrosome® is disrupted and metabolized in the phagolysosome will be dispersed among the residual apoptotic bodies which are subsequently phagocytosed by other macrophages. Therefore, fluorescent lipids may be detected in phagocytic cells which never phagocytosed Clodrosome® especially when FACS or fluoroscopy are utilized to detect fluorescent cells (FACS) or fluorescence levels in a tissue homogenate (fluoroscopy). Another potential artifact arises from fluorescent lipid remaining in the extracellular “garbage”, which has not yet been cleared by other phagocytes, generating a high background fluorescence. However, experienced confocal microscopist may be able to differentiate between the punctate fluorescence, resulting from fluorescent intact liposomes versus the more diffuse fluorescence characteristic of disrupted liposomes and some have successfully used fluorescent clodronate liposomes to visualize the cellular location of these liposomes by confocal microscopy in vivo [1]. A further complicating factor is that published data varies widely as to exactly when clodronate liposomes begin to induce apoptosis in macrophages. Mönkönnnen et al. show that macrophage death is measurable within the first hour after clodronate liposome treatment on RAW264 cells in vitro [2], while many others have reported no signs of macrophage apoptosis until several hours after treatment in vivo. The variability in the data is likely due to different liposomal formulations of clodronate as well as the vastly different experimental conditions. Therefore, as with most biological studies, especially those involving liposomes, the amount of time between treating the animal or cells with clodronate liposomes and the onset of apoptosis will need to be established in each experimental model. If the nature of the research demands that Clodrosome® be tracked rather than the control, Encapsula can provide DiI-labelled Clodrosome® upon request, and assuming that the Clodrosome® distribution can definitively be assessed prior to the onset of apoptosis, clear and valid data on the biodistribution of fluorescent Clodrosome® should be obtainable. Still, for most purposes, Fluoroliposome® (fluorescent control liposomes) will provide the required data with far fewer potential artifacts.
- When monitoring monocyte uptake in vivo in normal animals, the circulating monocytes may “disappear” or show reduced counts within the first 2 h post-injection due to margination of the monocytes post-liposome phagocytosis. These cells will re-enter the circulation within a few hours. Sunderkötter et al. demonstrate this phenomenon and discuss the behavior in detail. Also consider that circulating monocytes have a lifetime of about 24 h so labeled monocytes will be continually leaving the circulation, even in normal animals, due to aging of the monocytes [3].
- Liposomes may settle when left undisturbed for more than a few hours. Immediately prior to use, in order to ensure a homogeneous liposome suspension, slowly invert the vial several times until the suspension appears homogeneous by visual inspection. Vigorous or erratic shaking will not damage the liposomes but may induce foaming and bubble formation making it more difficult to accurately measure the desired dosage.
- If the personnel performing intravenous injections are not experienced in or familiar with, precautions for injecting larger volumes (~10% animal weight in ml), viscous liquids or particulate suspensions, consider having extra animals available in case serious injection-related adverse events occur. Dose control animals first to become familiar with large volume injections.
- When dosing intravenously, use standard precautions for dosing larger volumes to animals including the following: a) Warm product to room temperature prior to dosing. b) Ensure that all air bubbles are removed from the syringe prior to dosing; intravenous injection of air bubbles may result in air emboli which can kill or seriously injure animals. c) Inject product at a slow, steady rate of no more than 1 ml/min; decrease infusion rate if animals display any atypical reactions such as unusual agitation.
- Infusion-related adverse reactions usually involve the animal gasping for air or other seizure-like movements. Animals often recover with no apparent permanent injury, but any potential effects on experimental results must be assessed by the researcher.
- Liposomes should be kept at 4°C and NEVER be frozen.
Dosage
Appearance
Fluoroliposome®-DiO is a yellow liquid suspension made of large micron size multilamellar liposomes. Due to their large size, some liposomes might settle to the bottom of the vial. If left sitting idle in the refrigerator, Fluoroliposome®-DiO will phase separate and form pellets in the bottom of the vial, leaving a clear solution on top. Therefore, the vial should be shaken to form a homogeneous solution prior to use.
Educational Videos
Ordering/Shipping Information
- All liposome based formulations are shipped on blue ice at 4°C in insulated packages using overnight shipping or international express shipping.
- Liposomes should NEVER be frozen. Ice crystals that form in the lipid membrane can rupture the membrane, change the size of the liposomes and cause the encapsulated drug to leak out. Liposomes in liquid form should always be kept in the refrigerator.
- Clients who order from outside of the United States of America are responsible for their government import taxes and customs paperwork. Encapsula NanoSciences is NOT responsible for importation fees to countries outside of the United States of America.
- We strongly encourage the clients in Japan, Korea, Taiwan and China to order via a distributor. Tough customs clearance regulations in these countries will cause delay in custom clearance of these perishable formulations if ordered directly through us. Distributors can easily clear the packages from customs. To see the list of the distributors click here.
- Clients ordering from universities and research institutes in Australia should keep in mind that the liposome formulations are made from synthetic material and the formulations do not require a “permit to import quarantine material”. Liposomes are NOT biological products.
- If you would like your institute’s FedEx or DHL account to be charged for shipping, then please provide the account number at the time of ordering.
- Encapsula NanoSciences has no control over delays due to inclement weather or customs clearance delays. You will receive a FedEx or DHL tracking number once your order is confirmed. Contact FedEx or DHL in advance and make sure that the paperwork for customs is done on time. All subsequent shipping inquiries should be directed to Federal Express or DHL.
Storage and Shelf Life
Storage
Fluoroliposome® products should always be stored at in the dark at 4°C, except when brought to room temperature for brief periods prior to animal dosing. DO NOT FREEZE. ENS is not responsible for results generated by frozen product.
Shelf Life
Fluoroliposome® products are made on daily basis. The batch that is shipped is manufactured on the same day. It is advised to use the products within 60 days of the manufacturing date.
References and background reading
1. Polfliet MM, Goede PH, van Kesteren-Hendrikx EM, van Rooijen N, Dijkstra CD, van den Berg TK. A method for the selective depletion of perivascular and meningeal macrophages in the central nervous system. J. Neuroimmunol. 2001 Jun 1;116(2):188–95.
2. Mönkkönen J, Liukkonen J, Taskinen M, Heath TD, Urtti A. Studies on liposome formulations for intra-articular delivery of clodronate. Journal of Controlled Release. 1995 Aug;35(2–3):145–54.
3. Sunderkötter C, Nikolic T, Dillon MJ, van Rooijen N, Stehling M, Drevets DA, Leenen P. Subpopulations of Mouse Blood Monocytes Differ in Maturation Stage and Inflammatory Response. J Immunol. 2004 Apr 1;172(7):4410–7.
4. Nagai H, Kuwahira I, Schwenke DO, Tsuchimochi H, Nara A, Ogura S, Sonobe T, Inagaki T, Fujii Y, Yamaguchi R, Wingenfeld L. Pulmonary macrophages attenuate hypoxic pulmonary vasoconstriction via β3AR/iNOS pathway in rats exposed to chronic intermittent hypoxia. PLoS One. 2015 Jul 1;10(7):e0131923.
5. Zhu Y, Soderblom C, Krishnan V, Ashbaugh J, Bethea JR, Lee JK. Hematogenous macrophage depletion reduces the fibrotic scar and increases axonal growth after spinal cord injury. Neurobiology of disease. 2015 Feb 28;74:114-25.
6. Yun MH, Davaapil H, Brockes JP. Recurrent turnover of senescent cells during regeneration of a complex structure. Elife. 2015;4:e05505.
7. Arwert EN, Harney AS, Entenberg D, Wang Y, Sahai E, Pollard JW, Condeelis JS. A Unidirectional Transition from Migratory to Perivascular Macrophage Is Required for Tumor Cell Intravasation. Cell reports. 2018 May 1;23(5):1239-48.
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短截就是把枝条剪短,主要作用是促使其抽生新梢,增加分枝数目,以保证树势健状和正常结果。短截常用于骨干枝组修剪,结果枝组修剪,和树体局部更新复状。
短截按其长度可分为:
① 中短截:在一年生枝的中部短截,剪后萌发的顶端枝条,长势强,下部枝条长势弱。
② 重短截:剪去一年生枝的2/3。剪后萌发出的枝条较强状,一般用于主侧枝延长头修剪。
③ 重剪:剪去一年生枝的3/4-4/5,剪后萌发出的枝条长势强状,常用于发育枝作延长枝头和徒长果枝,中果枝的修剪。
④极重短截:剪去一年生枝的4/5以上,萌发后的枝条中庸偏状,常用于将发育枝和徒长枝培养结果枝组。
⑤留基部2芽剪:剪后萌发枝条较旺盛,常用于预备枝的修剪。对于幼龄树,树势较旺,以培养良好而牢固的树形结构,提早结果为主要目的,以轻短截,少疏间为主,从始果期到盛果期,主要使桃树多结果,并形成好的树形。
将来可能会出现基于免疫方法的试剂盒,不过因为H7N9是个新病毒,爆发至今不足一月,这么快时间还来不及生产抗体,所以目前的检测试剂盒只能是PCR检测试剂盒。
查到了:
4月7日,上海之江生物科技有限公司官方微博称,该公司成功研制禽流感H7N9(2013)核酸测定试剂盒(荧光PCR法),是针对国内此次H7N9病毒最早研制成功的产品,也是国内目前唯一供应的成品化试剂盒。
它一般在医院、制药企业使用。
试剂盒使用示例: 试剂盒的产生正是为了使实验人员能够摆脱繁重的试剂配制及优化过程,所以试剂盒中一般配备有相应的使用说明书
从1~2个T75培养皿的感染细胞培养液中纯化腺病毒
产品简介
本系列试剂盒可快速,高效地从腺病毒感染的细胞培养液中分离纯化腺病毒,相比于传统CsCl超速离心的腺病毒病毒纯化方式(需要24个小时才能完成纯化),本试剂盒可以在1个小时内完成病毒纯化,操作简单,快速,纯化率高,纯化到的病毒颗粒可直接用于下游实验,如细胞和动物感染。
产品特点
?操作简单,快速,可以在1个小时内完成病毒纯化,不依赖于超速离心操作。
?纯化效率高:小量纯化,可从1~2个T75瓶培养的细胞培养液中纯化病毒颗粒高达1x1012VPs
?每个纯化柱,可以重复利用一次,用于纯化相同种类的腺病毒。
保存条件
纯化柱和脱盐柱保存于4℃,其它组分室温保存。
试剂盒组分
Catalog#
V1160-01
Notes
Preps
10
MiniColumns
5
Canbeusedtwice
(Storeat4°C)
Press-OnCap
5
StoreatRT
DesaltingTube*
1
Canberegenarated
(Storeat4°C)
15mLCollectionTube
10
StoreatRT
10xWashBuffer
30mL
StoreatRT
2xElutionBuffer
30mL
StoreatRT
RegenerationBuffer
30mL
StoreatRT
联系方式:
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