BODIPY 505/515
(Synonyms: 4,4-二氟-1,3,5,7-四甲基-4-硼-3A,4A-二氮杂-S-茚烯) 目录号 : GC42960
BODIPY 505/515是一种亲脂性荧光染料，发出的荧光被广泛用于标记脂质滴（激发/发射：505/515纳米）。
Cas No.:21658-70-8
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
本方案仅提供一个指导，请根据您的具体需要进行修改。
1.染色液的制备
(1)配置储存液:使用DMSO溶解BODIPY 505/515，配置浓度为1-5mM的储存液。
注意:未使用的储存液分装后在-20℃或-80°C避光保存，避免反复冻融。
(2)配置工作液:用合适的缓冲液(如:无血清培养基或PBS)稀释储存液，配制浓度为1-5μM的工作液。
注意:请根据实际情况调整工作液浓度，现用现配。
2.细胞悬浮染色
(1)悬浮细胞：经4°C、1000g离心3-5分钟，弃去上清液，用PBS清洗两次，每次5分钟。
(2)贴壁细胞：使用PBS清洗两次，加入胰酶消化细胞，消化完成后经1000g离心3-5min。
(3)加入1mL的BODIPY 505/515工作溶液重悬细胞，37 °C避光孵育5-30分钟。不同细胞最佳孵育时间不同，请根据具体实验需求自行摸索。
(4)孵育结束后，经1000g离心5分钟，去除上清液，加入PBS清洗2-3次，每次5分钟。
(5)用预温的无血清细胞培养基或PBS重悬细胞，通过荧光显微镜或流式细胞术观察。
3.细胞贴壁染色
(1)在无菌盖玻片上培养贴壁细胞。
(2)从培养基中移走盖玻片，吸出过量的培养基，将盖玻片放在潮湿的环境中。
(3)从盖玻片的一角加入100μL的染料工作液，轻轻晃动使染料均匀覆盖所有细胞。
(4) 37 °C避光孵育5-30分钟。不同细胞最佳孵育时间不同，请根据具体实验需求自行摸索。
(5)孵育结束后吸弃染料工作液，使用预温的培养液清洗盖玻片2~3次。
4.显微镜检测：BODIPY 505/515的最大激发/发射光分别为505/515 nm。
注意事项：
①建议设置阳性对照，对照组细胞使用30μM油酸孵育8小时后进行后续实验；
②荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭；
为了您的安全和健康，请穿实验服并戴一次性手套操作。
BODIPY 505/515, a lipophilic fluorescence dye, emits fluorescence has been used extensively for lipid droplet labeling (Ex/Em: 505/515 nm) [1，2] . BODIPY 505/515 is compatible with epifluorescent, confocal, and two-photon microscopy, and flow cytometry, and can be used for live and fixed cell applications. Bodipy 505/515 is used for labeling a broad variety of lipids, e.g., phospholipids, fatty acids, cholesteryl esters, cholesterol, and ceramides
BODIPY 505/515是一种亲脂性荧光染料，发出的荧光被广泛用于标记脂质滴（激发/发射：505/515纳米）[1,2]。BODIPY 505/515适用于表观荧光、共聚焦和双光子显微镜、流式细胞术，并可用于活体和固定细胞应用。 Bodipy 505/515可用于标记各种类型的脂类，例如磷脂、脂肪酸、胆固醇酯、胆固醇和神经酰胺。
References:
[1]. Velmurugan, N., Sathishkumar, Y., Yim, S.S., et al. Study of cellular development and intracellular lipid bodies accumulation in the thraustochytrid Aurantiochytrium sp. KRS101. Bioresour. Technol. 161, 149-154 (2014).
[2]. Rumin, J., Bonnefond, H., Saint-Jean, B., et al. The use of fluorescent Nile red and BODIPY for lipid measurement in microalgae. Biotechnol. Biofuels 8, 42 (2015).
[3]. Elle I C, Olsen L C B, Pultz D, et al. Something worth dyeing for: molecular tools for the dissection of lipid metabolism in Caenorhabditis elegans[J]. FEBS letters, 2010, 584(11): 2183-2193.
| Cas No. | 21658-70-8 | SDF | |
| 别名 | 4,4-二氟-1,3,5,7-四甲基-4-硼-3A,4A-二氮杂-S-茚烯 | ||
| 化学名 | (T-4)-[2-[(3,5-dimethyl-2H-pyrrol-2-ylidene-κN)methyl]-3,5-dimethyl-1H-pyrrolato-κN]difluoro-boron | ||
| Canonical SMILES | [F-][B+3]([N-]1C2=C(C)C=C1C)([N](C(C(C)=C3)=C2)=C3C)[F-] | ||
| 分子式 | C13H15BF2N2 | 分子量 | 248.1 |
| 溶解度 | 1.7mg/ml in DMSO; 25mg/ml in DMF | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 4.0306 mL | 20.1532 mL | 40.3063 mL |
| 5 mM | 0.8061 mL | 4.0306 mL | 8.0613 mL |
| 10 mM | 0.4031 mL | 2.0153 mL | 4.0306 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Detection and quantitation of lipid in the microalga Tetraselmis subcordiformis (Wille) Butcher with BODIPY 505/515 staining
Bioresour Technol 2013 Jan;127:386-90.PMID:23138061DOI:10.1016/j.biortech.2012.09.068.
BODIPY 505/515, a lipophilic bright green fluorescent dye was tested for lipid detection in the microalga Tetraselmis subcordiformis. A concentration of 0.28 μg ml(-1) and staining for 6 min was optimal. Lipid bodies stained with BODIPY505/515 had a characteristic green fluorescence. Their volumes were determined using the sphere volume formula. Lipid accumulation under different nitrogen concentrations was analyzed. With an increase in NaNO(3) concentration from 0 to 240 mg L(-1), the maximum algal concentration increased from 8.23 ± 0.62 (× 10(5) cells ml(-1)) to 1.61 ± 0.13 (×10(6) cells ml(-1)), while the maximum volume of intracellular neutral lipid decreased from 9.78 ± 1.77 μm(3) cell(-1) to 6.00 ± 0.59 μm(3) cell(-1). A comparison of the lipid contents measured by BODIPY 505/515 staining and the gravimetric method showed a positive correlation coefficient of R(2) = 0.93. BODIPY 505/515 staining is a promising method in lipid quantitation in T. subcordiformis.
Confocal microscopic analysis of morphogenetic movements
Methods Cell Biol 1999;59:179-204.PMID:9891361DOI:10.1016/s0091-679x(08)61826-9.
Confocal microscopy is an excellent means of imaging cellular dynamics within living zebrafish embryos because it provides a means of optically sectioning tissues that have been labeled with specific fluorescent probe molecules. In order to study genetically encoded patterns of cell behavior that are involved in the formation of germ layers and various organ primordia, it is possible to vitally stain an entire zebrafish embryo with one or more fluorescent probe molecules and then examine morphogenetic behaviors within specific cell populations of interest using time-lapse confocal microscopy. There are two major advantages to this "bulk-labeling" approach: (1) the applied fluorescent probe (a contrast-enhancing agent) allows all of the cells within an intact zebrafish embryo to be rapidly stained; (2) the morphogenetic movements and shape changes of hundreds of cells can then be examined simultaneously in vivo using time-lapse confocal microscopy. The neutral fluorophore BODIPY 505/515 and its sphingolipid-derivative Bodipy-C5-ceramide are particularly useful, nonteratogenic vital stains for imaging cellular dynamics in living zebrafish embryos. These photostable fluorescent probes (when applied with 2% DMSO) percolate through the enveloping layer epithelium of the embryo, and localize in yolk-containing cytoplasm and interstitial space, respectively, owing to their different physiochemical characteristics. Bodipy-ceramide, for instance, remains highly localized to interstitial fluid once it accumulates within a zebrafish embryo, allowing the boundaries of deep cells to be clearly discerned throughout the entire embryo. Through the use of either of these fluorescent vital stains, it is possible to rapidly convert a developing zebrafish embryo into a strongly fluorescent specimen that is ideally suited for time-lapse confocal imaging. For zebrafish embryos whose deep cells have been intentionally "scatter-labeled" with fluorescent lineage tracers (e.g., fluorescent dextrans), sequential confocal z-series (i.e., focus-throughs) of the embryo can be rendered into uniquely informative 3D time-lapse movies using readily available image-processing programs. Similar time-lapse imaging, combined with rapidly advancing computer-assisted visualization techniques, may soon be applied to study the dynamics of GFP-fusion proteins in vivo, as well as other types of synthetic probe molecules designed to reveal the cytological processes associated with the patterning and morphological transformations of the zebrafish's embryonic tissues.
Advances in techniques for assessment of microalgal lipids
Crit Rev Biotechnol 2017 Aug;37(5):566-578.PMID:27417693DOI:10.1080/07388551.2016.1206058.
Microalgae are a varied group of organisms with considerable commercial potential as sources of various biochemicals, storage molecules and metabolites such as lipids, sugars, amino acids, pigments and toxins. Algal lipids can be processed to bio-oils and biodiesel. The conventional method to estimate algal lipids is based on extraction using solvents and quantification by gravimetry or chromatography. Such methods are time consuming, use hazardous chemicals and are labor intensive. For rapid screening of prospective algae or for management decisions (e.g. decision on timing of harvest), a rapid, high throughput, reliable, accurate, cost effective and preferably nondestructive analytical technique is desirable. This manuscript reviews the application of fluorescent lipid soluble dyes (Nile Red and BODIPY 505/515), nuclear magnetic resonance (NMR), Raman, Fourier transform infrared (FTIR) and near infrared (NIR) spectroscopy for the assessment of lipids in microalgae.
Evaluation of intracellular lipid bodies in Chlamydomonas reinhardtii strains by flow cytometry
Bioresour Technol 2013 Jun;138:30-7.PMID:23612159DOI:10.1016/j.biortech.2013.03.078.
A comparative study of Chlamydomonas reinhardtii wild type CC124 and a cell wall-less mutant sta6-1 is described using FACS in conjunction with two different lipophilic fluorescent dyes, Nile Red and BODIPY 505/515. The results indicate that BODIPY 505/515 is more effective for the vital staining of intracellular lipid bodies and single cell sorting than Nile Red. While BODIPY 505/515 stained cells continued to grow after single cell sorting using FACS, Nile Red stained cells failed to recover from sorting. In addition, a comprehensive study was performed to establish a quantitative baseline for future studies for either lipid accumulation and/or microalgal growth by measuring various parameters such as cell count, size, fatty acid contents/composition, and optical/confocal images of the wild type and mutant.
Visualizing "green oil" in live algal cells
J Biosci Bioeng 2010 Feb;109(2):198-201.PMID:20129108DOI:10.1016/j.jbiosc.2009.08.004.
We report here that BODIPY 505/515, a green lipophilic fluorescent dye, serves as an excellent vital stain for the oil-containing lipid bodies of live algal cells. BODIPY 505/515 vital staining can be used in combination with fluorescent activated cell sorting to detect and isolate algal cells possessing high lipid content.