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DiI碘化物

DiI, DiO, DiD and DiR dyes are a family of lipophilic fluorescent stains for labeling membranes and other hydrophobic structures. The fluorescence of these environment-sensitive dyes is greatly enhanced when incorporated into membranes or bound to lipophilic biomolecules such as proteins although they are weakly fluorescent in water. They have high extinction coefficients, polarity-dependent fluorescence and short excited-state lifetimes. Once applied to cells, these dyes diffuse laterally within the cellular plasma membranes, resulting in even staining of the entire cell at their optimal concentrations. The distinct fluorescence colors of DiI (orange fluorescence), DiO (green fluorescence), DiD (red fluorescence) and DiR (deep red fluorescent) provide a convenient tool for multicolor imaging and flow cytometric analysis of live cells. DiO and DiI can be used with standard FITC and TRITC filters respectively. Among them DiI and its analogs are most frequently used since they usually exhibit very low cell toxicity. In addition, DiI is widely used for determining lipoproteins such as LDL and HDL.

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1000

产品编号

22101-10mg/1198元

所属分类

Fluorescence reagents and more

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

产品描述

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DiD，DiO，DiI，DiR and DiS

lipophilic fluorescent stains for labeling membranes

DiI iodide [1,1-Dioctadecyl-3,3,3,3- tetramethylindocarbocyanine iodide]

Cat#	Size	Price	MW	Abs	Em	Soluble in	Storage
22101	10mg	$95	961.32	549 nm	565 nm	DMSO	F/D/L

Features and Biological Applications

DiI, DiO, DiD and DiR dyes are a family of lipophilic fluorescent stains for labeling membranes and other hydrophobic structures. The fluorescence of these environment-sensitive dyes is greatly enhanced when incorporated into membranes or bound to lipophilic biomolecules such as proteins although they are weakly fluorescent in water. They have high extinction coefficients, polarity-dependent fluorescence and short excited-state lifetimes. Once applied to cells, these dyes diffuse laterally within the cellular plasma membranes, resulting in even staining of the entire cell at their optimal concentrations. The distinct fluorescence colors of DiI (orange fluorescence), DiO (green fluorescence), DiD (red fluorescence) and DiR (deep red fluorescent) provide a convenient tool for multicolor imaging and flow cytometric analysis of live cells. DiO and DiI can be used with standard FITC and TRITC filters respectively. Among them DiI and its analogs are most frequently used since they usually exhibit very low cell toxicity. In addition, DiI is widely used for determining lipoproteins such as LDL and HDL.

References

1. Heinrich L, Freyria AM, Melin M, Tourneur Y, Maksoud R, Bernengo JC, Hartmann DJ. (2006) Confocal laser scanning microscopy using dialkylcarbocyanine dyes for cell tracing in hard and soft biomaterials. J Biomed Mater Res B Appl Biomater.

2. Higashide T, Kawaguchi I, Ohkubo S, Takeda H, Sugiyama K. (2006) In vivo imaging and counting of rat retinal ganglion cells using a scanning laser ophthalmoscope. Invest Ophthalmol Vis Sci, 47, 2943.

3. Kalchenko V, Shivtiel S, Malina V, Lapid K, Haramati S, Lapidot T, Brill A, Harmelin A. (2006) Use of lipophilic near-infrared dye in whole-body optical imaging of hematopoietic cell homing. J Biomed Opt, 11, 050507.

4. Wang G, Anrather J, Glass MJ, Tarsitano MJ, Zhou P, Frys KA, Pickel VM, Iadecola C. (2006) Nox2, Ca2+, and protein kinase C play a role in angiotensin II-induced free radical production in nucleus tractus solitarius. Hypertension, 48, 482.

5. Wertz A, Rossler W, Obermayer M, Bickmeyer U. (2006) Functional neuroanatomy of the rhinophore of Aplysia punctata. Front Zool, 3, 6.

6. Harrison TA, Perry KM, Hoover DB. (2005) Regional cardiac ganglia projections in the guinea pig heart studied by postmortem DiI tracing. Anat Rec A Discov Mol Cell Evol Biol, 285, 758.

7. Huesa G, Anadon R, Yanez J. (2003) Afferent and efferent connections of the cerebellum of the chondrostean Acipenser baeri: a carbocyanine dye (DiI) tracing study. J Comp Neurol, 460, 327.

8. Kiyohara Y, Endo K, Ide C, Mizoguchi A. (2003) A novel morphological technique to investigate a single climbing fibre synaptogenesis with a Purkinje cell in the developing mouse cerebellum: DiI injection into the inferior cerebellar peduncle. J Electron Microsc (Tokyo), 52, 327.

9. Levai O, Strotmann J. (2003) Projection pattern of nerve fibers from the septal organ: DiI-tracing studies with transgenic OMP mice. Histochem Cell Biol, 120, 483.

10. Pavlidis M, Stupp T, Naskar R, Cengiz C, Thanos S. (2003) Retinal ganglion cells resistant to advanced glaucoma: a postmortem study of human retinas with the carbocyanine dye DiI. Invest Ophthalmol Vis Sci, 44, 5196.

表1. DiD，DiO，DiI，DiR，DiS的化学性质

目录号	产品名称	规格	分子量	Ex/Em	推荐滤光器*
DiD
22033	DiD labeling solution	10 mL	959.91	644/663 nm	XF47-Omega, 31023-Chroma
DiO
22040	DiOC7(3) iodide	100 mg	600.57	482/504 nm	XF23-Omega, 31001-Chroma
22046	DiOC6(3) iodide	100 mg	572.52	482/504 nm	XF23-Omega, 31001-Chroma
22066	DiO perchlorate	100 mg	881.7	484/501nm	XF23-Omega, 31001-Chroma
DiI
22035	DiIC12(3) perchlorate	100 mg	765.55	549/565 nm	XF32-Omega, 31002-Chroma
22044	DiIC16(3) perchlorate	100 mg	877.76	549/565 nm	XF32-Omega, 31002-Chroma
22052	DiIC18(3)-DS	50 mg	993.53	555/570 nm	XF32-Omega, 31002-Chroma
22054	DiIC18(5)-DS	50 mg	1019.57	650/670 nm	XF47-Omega, 31023-Chroma
22056	DiIC1(5) iodide	100 mg	510.45	638/658 nm	XF47-Omega, 31023-Chroma
22101	DiI iodide	100 mg	961.32	549/565 nm	XF32-Omega, 31002-Chroma
22102	DiI perchlorate	100 mg	933.87	549/565 nm	XF32-Omega, 31002-Chroma
22103	DiI triflate	100 mg	983.48	549/565 nm	XF32-Omega, 31002-Chroma
DiR**
22070	DiR iodide	10 mg	1013.39	748/780 nm	XF112-Omega, 41009-Chroma
DiS
22073	DiSC2(3)	25 mg	492.44	560/571 nm	XF32-Omega, 31002-Chroma
22076	DiSC3(5)	100 mg	546.53	660/675 nm	XF47-Omega, 31023-Chroma

* Omega滤光器由Omega公司提供（www. omegafilters.com）；Chroma滤光器由Chroma公司提供（www. chroma.com）。

**DiR的发射波长肉眼不可见，所以需要配备CCD镜头或其他的近红外检测仪器。

使用方法

1. DiD，DiO，DiI，DiR和DiS细胞膜染色液制备

（1）配置DMSO或EtOH 储存液：储存液用 DMSO或EtOH配置浓度1～5 mM。

注意：未使用的储存液保存在-20°C，避免反复冻融。

（2）工作液制备：用合适的缓冲液（如：无血清培养基，HBSS或PBS）稀释储存液，配制浓度为1～5 µM的工作液。

注意：工作液的最终浓度是根据不同细胞和实验的经验来配制。可以从推荐浓度的十倍以上寻找最佳条件。

2. 悬浮细胞染色

（1）悬浮细胞密度为 1 × 106/mL加入到工作液中。

（2）在37 ℃培养细胞 2～20分钟，不同的细胞最佳培养时间不同。

（3）染色细胞试管在1000～1500转离心5分钟。

（4）倾倒上清液，再次缓慢加入预温37℃的培养液。

（5）重复（3），（4）步骤两次以上。

3. 粘壁细胞的染色

（1）使粘壁的细胞在无菌实验室培养。

（2）从培养基中移走盖玻片，吸走过量培养液，将盖玻片放在潮湿的环境中。

（3）在盖玻片的一角加入100μL的染料工作液，轻轻晃动使染料均匀覆盖所有细胞。

（4）在37 ℃培养细胞 2～20分钟，不同的细胞最佳培养时间不同。

（5）吸干染料工作液，用培养液洗盖玻片2～3次，每次用预温的培养基覆盖所有细胞，培养5～10分钟，然后吸干培养基。

4. 显微镜检测

（1）DiD，DiO，DiI，DiR和DiS滤光器的选择参见表1。

（2）多色染料的同时检测，滤光器按照以下设定：

a) DiI和DiO＝Omega XF52，Chroma 51004；

b) DiI和DiD＝Omega XF92，Chroma 51007；

c) DiI，DiO和DiD＝Omega XF93，Chroma 61005

5. 流式细胞仪的检测

DiD，DiO，DiI，DiR和DiS 染色的细胞可以分别用经典的FL1，FL2，FL3和FL4流式细胞仪检测。

储存条件：-20℃干燥避光保存，有效期一年。

特别提醒：

1) 本公司所有产品仅限于专业人员用于生命科学研究，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅。

2) 本公司所有产品必须由合格专业技术人员操作同时佩戴口罩/手套/实验服并遵守生物实验室安全操作规程！

产品参数

Name	DiI iodide
CAT#	22101-10mg	CAS#
Storage#	−20°C Sealed & desiccated & Minimized light exposure	Shelf Life#	24 months
Ex(nm)#	549	Em(nm)#	565
MW#	961.32	Solvent#	DMSO

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DiI碘化物

DiI iodide [1,1-Dioctadecyl-3,3,3,3- tetramethylindocarbocyanine iodide]

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