计算溶液所需的质量、体积或浓度。
荧光法脂滴检测试剂盒 (BODIPY 493/503, 绿色)
有货
为什么选择此级别
BioReagent,即用型,无菌,适用于荧光分析 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
避光,-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
脂滴（Lipid droplets, LDs），也可称为油体，区别于脂质体，是真核细胞内重要的功能性细胞器。其结构由磷脂单分子层及内部中性脂疏水核心构成，核心成分主要为甘油三酯与胆固醇酯，脂滴尺寸差异显著，直径范围为40 nm～100 μm。作为细胞内中性脂质的主要储存位点，脂滴可沿细胞骨架运动，并与多种细胞器发生相互作用，广泛参与机体能量储存、脂质代谢与储存、膜转运、蛋白降解、信号传导等生物学过程。同时，脂滴结构与功能异常，与肥胖、脂肪肝、心血管疾病、糖尿病等多种代谢性疾病密切相关。
本绿色荧光脂滴检测试剂盒基于BODIPY 493/503特异性荧光探针开发，可精准检测细胞内脂滴。其中，BODIPY 493/503探针在水及极性溶剂中几乎无荧光信号，与甘油三酯等中性脂质结合后可激发出明亮的绿色荧光，激发/发射波长（Ex/Em）为493 nm/503 nm。配套的Hoechst 33342为细胞膜通透性蓝色荧光染料，可嵌入细胞双链DNA并产生强烈蓝色荧光（Ex/Em=346 nm/460 nm），适用于细胞核定位染色，可实现脂滴与细胞核的双重定位观察。
本试剂盒核心组分包括：BODIPY 493/503 (1000×)、细胞核染色试剂Hoechst 33342 (1000×) 及检测缓冲液。产品适配荧光显微镜、流式细胞仪、荧光酶标仪等主流荧光检测设备，可清晰观测细胞内脂滴的分布形态与数量变化。
试剂盒分两种规格，不同检测场景下的可检测次数如下：
①96孔板检测：单孔染色体系100 μL，100-1000T规格可检测1000次，500-5000T规格可检测5000次；
②流式细胞仪检测：单样品检测体系0.5 mL，100-1000T规格可检测200次，500-5000T规格可检测1000次；
③6孔板贴壁细胞染色：单孔染色体系1 mL，100-1000T规格可检测100次，500-5000T规格可检测500次。
产品组成
| 组分货号 | 组分名称 | 物理外观 | 100-1000T | 500-5000T | 储存 |
| F1527962A | BODIPY 493/503 (1000×) | 液体 | 0.1mL | 0.5mL | -20℃.避光 |
| F1527962B | Hoechst 33342 (1000×) | 液体 | 0.1mL | 0.5mL | -20℃. 避光 |
| F1527962C | 检测缓冲液 | 液体 | 100mL | 500mL | -20℃. 避光 |
使用方法
（一）阳性对照的诱导方法
油酸 (阿拉丁货号：O1456059)和油酸钠 BSA 高脂细胞添加剂 (阿拉丁货号：S767993)均可以诱导培养细胞内脂滴生成，作为阳性对照。
油酸 (阿拉丁货号：O1456059)使用具体方法如下：
1. 37ºC 条件下加热油酸，使其完全成为液体。
2. 取 48 μL 的油酸，加入到 952 μL 的 DMSO 中，充分混匀，配制成 150 mM 的油酸贮存液。贮存液可以短期保存在 4ºC，使用时 37ºC 条件下加热混匀后即可使用。
3. 配制含油酸的完全培养液：使用细胞的完全培养液按 500：1 稀释油酸贮存液，使油酸终浓度为300 μM。油酸具有一定的细胞毒性，因细胞而异，可以根据不同细胞状况调整油酸的使用终浓度。
4. 待检细胞加入含油酸的完全培养液，37ºC 培养过夜。一般情况下，次日即可观察到囊泡状脂滴。
油酸钠 BSA 高脂细胞添加剂 (阿拉丁货号：S767993)使用具体方法如下：
1. 将油酸钠 BSA 高脂细胞添加剂完全融化后，使用细胞的完全培养液稀释至 300 μM-500 μM。
2. 待检细胞加入含油酸钠的完全培养液，37ºC 培养 24-48h，即可观察到囊泡状脂滴。
（二）染色工作液的配制（以 1000×稀释为例）
由于不同细胞种类、细胞密度的染色条件不同，建议自行摸索 BODIPY 493/503 和 Hoechst 33342的最适浓度。一般来说，满足信号足够的前提下，尽可能选择最低浓度的染料剂量。
根据样品数量和每个样品所需工作液的体积，计算出染色工作液的总体积。对于 6、12、24、96 孔板，每孔检测工作液的用量分别为 0.5~1mL、200~500μL、100~250μL 和 50~100μL；对于流式细胞样品，每个样品的染色工作液的体积为 0.5mL。
下述按照 1:1000 稀释倍数/1mL 举例：
取出 BODIPY 493/503 (1000×) 、Hoechst 33342 (1000×) 和检测缓冲液完全融化，平衡至室温。参考下表配制适量的染色液，涡旋震荡混匀制成染色工作液。
| 试剂 | 体积 |
| BODIPY 493/503 (1000×) | 1 μL |
| Hoechst 33342 (1000×) | 1 μL |
| 检测缓冲液（Assay Buffer） | 998 μL |
| 总计 | 1 mL |
配制好的染色工作液必须一次使用完毕，不建议冻存或 4ºC 保存后继续使用。染色液中 BODIPY493/503 的最终浓度可以根据染色效果在 0.2× ~ 2×之间适当调整。
（三）荧光显微镜检测
1. 接种培养：将细胞接种于 96 孔板等多孔板、细胞培养皿中或者细胞爬片上，按实验设计对细胞进行一定处理。
2. 洗涤：对于贴壁细胞，吸除培养液，用 PBS 洗涤细胞 2 次；对于悬浮细胞，250-1000 ×g 室温离心 5 min，吸除上清，用 PBS 洗涤 2 次。
3. 固定（选做）：加入 4%多聚甲醛固定液室温固定 10-15 min。固定结束后，用 PBS洗涤 1-2 次。
【注 1】：BODIPY 493/503 和 Hoechst 33342 适用于活细胞染色，也适用于固定后细胞的染色。但为了获得最佳的脂滴形态染色效果，在实验允许的情况下，建议将样品固定后再进行后续染色步骤。
【注 2】：由于醇类能够溶解脂质，因此请使用醛类固定液进行固定。
【注 3】：如果需要进行免疫染色而进行细胞通透，须避免使用含 Triton X-100 或 Tween-20 等去垢剂的通透液，而使用不会溶解细胞膜的去垢剂如含 Saponin 或 Digitonin 的通透液，但仍然可能会一定程度的影响脂滴的形态。
4. 染色。吸除孔板内原有 PBS，加入适当体积的染色工作液。通常 96 孔板每孔加入 100μL，24 孔板每孔加入 250μL，12 孔板每孔加入 500μL，6 孔板每孔加入 1mL。室温避光孵育 10-20min。不同的细胞最佳孵育时间有所不同，以 10min 作为初始孵育时间，后续可以根据实际染色效果对染色时间进行适当调整和优化，以得到更加理想的染色效果。
5. 检测。孵育结束后，用 PBS 洗两遍。随后，使用荧光显微镜观察，BODIPY 493/503 观察绿色荧光；Hoechst 33342 观察蓝色荧光。如有需要，也可进一步进行其它荧光的复染，注意整个过程均需注意避光操作。
【注 4】：使用荧光显微镜拍照时，为了减少荧光淬灭，尽可能降低荧光显微镜的激发光强度，缩短拍照时间。
（四）流式细胞仪检测
1. 细胞准备：贴壁细胞胰酶消化后用培养液重悬，并用 PBS 洗涤一次；悬浮细胞 400×g 室温离心
5min，弃上清，用 PBS 洗涤 1 次。每个样品推荐的细胞用量为 106个细胞。
2. 固定（选做）：加入 4%多聚甲醛固定液室温固定 10-15 min。
【注 1，2，3】：同上。
3. 洗涤：400 ×g 室温离心 5 min，吸除上清，用 PBS 洗涤 1-2 次。
4. 染色：对于上一步骤的细胞沉淀，除背景对照管外，其余每管加入 0.5 mL 染色工作液，重悬为单细胞悬液，室温避光孵育 10-15 min。孵育结束后，400 ×g 室温离心 5 min，弃上清，用 PBS 洗涤 2 次。
5. 检测：400 ×g 室温离心 5 min 沉淀细胞。吸净液体后，每个样品加入 0.5 mL 检测缓冲液重悬细胞后用流式细胞仪检测（检测时参考探针光谱特征选择合适的检测条件，例如Ex/Em=488/510 nm。）。
注意整个过程均需避光操作。染色后，尽量在 1 h 内进行流式细胞仪检测和分析。
【注 5】：使用仅含检测缓冲液并且未经染色的细胞样品用于流式细胞仪的阴性对照设置。
【注 6】：由于流式检测比较灵敏，使用的荧光探针浓度可能要比荧光显微镜检测时要低，此时可根据细胞类型和实际染色情况对 BODIPY 493/503 的稀释倍数进行适当调整。
（五）荧光酶标仪检测
1. 接种培养。将细胞接种于 96 孔黑色多孔板中，每孔的细胞数需要控制在 100-10000 个，通常宜在 2000-5000 个范围内。按实验设计对细胞进行一定处理。
2. 固定（选做）：加入 4%多聚甲醛固定液室温固定 10-15min。
【注 1，2，3】：同上。
3. 洗涤。对于贴壁细胞，吸除孔内原有液体，用 PBS 洗涤细胞 2 遍；对于悬浮细胞，250-1000 ×g室温离心 5 min，吸除上清，用 PBS 洗涤 2 遍。
4. 染色。加入适当体积的染色工作液，通常 96 孔板每孔加入 100 μL。室温避光孵育 10-20 min。不同的细胞最佳孵育时间有所不同，以 10 min 作为初始孵育时间，后续可以根据实际染色效果对染色时间进行适当调整和优化，以得到更加理想的染色效果。
5. 检测。孵育结束后，用 PBS 洗涤细胞 2 遍。随后用荧光酶标仪检测（参考探针光谱特征，选取合适的波长进行读板，例如 Ex/Em=488/510 nm。具体根据酶标仪的特点选择，也不一定需要选用最大激发光/发射光波长来检测）。
注意事项
1. 第一次使用请将本品分装并于-20℃冻存，避免反复冻融。建议根据单次用量分装密封保存。
2. BODIPY 493/503 的工作浓度、细胞量、孵育温度和时间等可根据所使用的细胞进行适当摸索和优化，例如 BODIPY 493/503 的浓度可在 0.2× ~ 2×之间适当调整。
3. BODIPY 493/503 在激光照射下易发生淬灭，因此需要在保证荧光亮度的前提下尽可能降低染料使用浓度，降低荧光显微镜激发光强度，缩短拍照时间。
4. 为获得准确试验结果，建议样品在染色后 1 小时内进行分析。
5. 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。
6. 检测缓冲液应在无菌环境中取用，否则可能会被微生物污染而影响使用效果，甚至无法继续使用。检测缓冲液可在短期内暂存 2-8ºC，一个月内有效。
7. 本产品仅限于专业人员的科学研究用，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内。
8. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Lipid droplets (LDs), also referred to as oil bodies and distinct from liposomes, are essential functional organelles in eukaryotic cells. They consist of a phospholipid monolayer surrounding a hydrophobic core of neutral lipids, whose major constituents are triglycerides and cholesteryl esters. Lipid droplets vary greatly in size, with diameters ranging from 40 nm to 100 μm. As the primary intracellular storage compartment for neutral lipids, lipid droplets migrate along the cytoskeleton and interact with multiple organelles. They participate extensively in biological processes including energy storage, lipid metabolism and storage, membrane trafficking, protein degradation, and signal transduction. Meanwhile, structural and functional abnormalities of lipid droplets are closely associated with diverse metabolic disorders such as obesity, fatty liver disease, cardiovascular diseases, and diabetes.
This Green Fluorescent Lipid Droplet Assay Kit is developed based on the specific fluorescent probe BODIPY 493/503 for accurate detection of intracellular lipid droplets. The BODIPY 493/503 probe exhibits virtually no fluorescence in water and polar solvents; upon binding to neutral lipids such as triglycerides, it emits bright green fluorescence with excitation/emission wavelengths (Ex/Em) of 493 nm/503 nm. The accompanying Hoechst 33342 is a cell‑permeable blue fluorescent dye that intercalates into double‑stranded cellular DNA to produce intense blue fluorescence (Ex/Em = 346 nm/460 nm). It is used for nuclear staining and enables dual‑color visualization of lipid droplets together with cell nuclei.
Key components of this kit include: BODIPY 493/503 (1000×), nuclear stain Hoechst 33342 (1000×), and assay buffer. Compatible with mainstream fluorescence‑detection instruments such as fluorescence microscopes, flow cytometers, and fluorescence microplate readers, the kit allows clear observation of the distribution, morphology, and quantitative changes of intracellular lipid droplets.
Two kit sizes are available. The number of assays for different experimental scenarios is shown below:
① 96‑well plate assay: staining volume per well = 100 μL; the 100‑1000T size provides 1000 assays, and the 500‑5000T size provides 5000 assays.
② Flow cytometry assay: staining volume per sample = 0.5 mL; the 100‑1000T size provides 200 assays, and the 500‑5000T size provides 1000 assays.
③ Adherent‑cell staining in 6‑well plates: staining volume per well = 1 mL; the 100‑1000T size provides 100 assays, and the 500‑5000T size provides 500 assays.
Product Components
| Component No. | Component Name | Appearance | 100-1000T | 500-5000T | Storage |
| F1527962A | BODIPY 493/503 (1000×) ） | Liquid | 0.1mL | 0.5mL | -20℃.Store in the dark. |
| F1527962B | Hoechst 33342 (1000×) | Liquid | 0.1mL | 0.5mL | -20℃.Store in the dark. |
| F1527962C | Assay Buffer | Liquid | 100mL | 500mL | -20℃.Store in the dark. |
Protocol
(I) Induction of Positive Control
Oleic acid (Aladdin Cat. No.: O1456059)and sodium oleate-BSA high-fat cell supplement ( Aladdin Cat. No.: S767993) can be used to induce intracellular lipid droplet formation in cultured cells as positive controls.
Procedure for Oleic Acid (Aladdin Cat. No.: O1456059)
1.Heat oleic acid at 37 °C until fully liquefied.
2.Add 48 μL oleic acid into 952 μL DMSO and mix thoroughly to prepare a 150 mM oleic acid stock solution. The stock can be stored short‑term at 4 °C; heat and mix at 37 °C before use.
3.Prepare complete medium containing oleic acid: dilute the oleic acid stock 500-fold with cell complete medium to a final oleic acid concentration of 300 μM. Oleic acid is cytotoxic in a cell-dependent manner; adjust the final working concentration according to cell type.
4.Incubate test cells with oleic acid-supplemented complete medium overnight at 37 °C. Vesicular lipid droplets are generally observable the next day.
Procedure for Sodium Oleate-BSA High-Fat Cell Supplement ( Aladdin Cat. No.: S767993)
1.Thaw the sodium oleate-BSA high-fat cell supplement completely, then dilute with cell complete medium to a final concentration of 300 μM–500 μM.
2.Incubate test cells with sodium oleate-supplemented complete medium at 37 °C for 24–48 h; vesicular lipid droplets can then be observed.
(II) Preparation of Staining Working Solution (example for 1000× dilution)
Optimal staining conditions vary by cell type and cell density; users are recommended to determine the best working concentrations of BODIPY 493/503 and Hoechst 33342. In general, use the lowest dye concentration that yields acceptable signal intensity.
Calculate the total volume of staining working solution according to sample number and required volume per sample. Recommended working volumes: 0.5–1 mL per well for 6-well plates, 200–500 μL per well for 12-well plates, 100–250 μL per well for 24-well plates, 50–100 μL per well for 96-well plates, and 0.5 mL per sample for flow cytometry.
The following example uses a 1:1000 dilution for a final volume of 1 mL:
Thaw BODIPY 493/503 (1000×), Hoechst 33342 (1000×) and Assay Buffer completely and equilibrate to room temperature. Prepare an appropriate amount of staining solution as referenced, then vortex to make staining working solution.
| Reagent | Volume |
| BODIPY 493/503 (1000×) | 1 μL |
| Hoechst 33342 (1000×) | 1 μL |
| Assay Buffer | 998 μL |
| Total | 1 mL |
Prepare only the required amount of staining working solution for single use; do not freeze or store at 4 °C for later reuse. The final concentration of BODIPY 493/503 can be adjusted within 0.2× ~ 2× according to staining performance.
(III) Detection by Fluorescence Microscope
1.Cell seeding and treatment: Seed cells in multi-well plates, culture dishes or coverslips, and treat cells according to the experimental design.
2.Washing: For adherent cells, aspirate medium and wash cells twice with PBS. For suspension cells, centrifuge at 250–1000 ×g for 5 min at room temperature, discard supernatant, and wash twice with PBS.
3.Fixation (optional): Add 4% paraformaldehyde fixative and incubate at room temperature for 10–15 min. After fixation, wash cells 1–2 times with PBS.
Note 1: BODIPY 493/503 and Hoechst 33342 are compatible with both live and fixed cells. For optimal lipid droplet morphology, fixation before staining is recommended if the experiment permits.
Note 2: Use aldehyde fixatives only; alcohols dissolve lipids.
Note 3: If permeabilization is required for immunostaining, avoid detergents such as Triton X‑100 or Tween‑20. Permeabilization reagents containing saponin or digitonin may be used, but can still alter lipid droplet morphology to some extent.
4.Staining: Remove PBS and add an appropriate volume of staining working solution. Typical volumes: 100 μL per well for 96-well plates, 250 μL per well for 24-well plates, 500 μL per well for 12-well plates, 1 mL per well for 6-well plates. Incubate at room temperature in the dark for 10–20 min. Optimal incubation time is cell-dependent; start with 10 min and adjust based on actual staining results.
5.Detection: Wash cells twice with PBS after incubation. Observe under a fluorescence microscope: green channel for BODIPY 493/503 and blue channel for Hoechst 33342. Additional counterstaining can be performed if needed; keep samples protected from light throughout the procedure.
Note 4: To reduce photobleaching during imaging, minimize excitation laser power and shorten exposure time.
(IV) Detection by Flow Cytometer
1.Cell preparation: Digest adherent cells with trypsin, resuspend in medium and wash once with PBS. Centrifuge suspension cells at 400 ×g for 5 min at room temperature, discard supernatant and wash once with PBS. A recommended cell number is 10⁶ cells per sample.
2.Fixation (optional): Add 4% paraformaldehyde fixative and incubate at room temperature for 10–15 min.
Notes 1, 2, 3: Same as above.
3.Washing: Centrifuge at 400 ×g for 5 min at room temperature, aspirate supernatant, and wash 1–2 times with PBS.
4.Staining: To the cell pellet (excluding the background control tube), add 0.5 mL staining working solution to prepare a single-cell suspension. Incubate at room temperature in the dark for 10–15 min. After incubation, centrifuge at 400 ×g for 5 min at room temperature, discard supernatant and wash twice with PBS.
5.Detection: Pellet cells by centrifugation at 400 ×g for 5 min at room temperature. Remove supernatant, resuspend cells in 0.5 mL Assay Buffer and analyze by flow cytometry (select appropriate detection parameters according to probe spectral properties, e.g., Ex/Em = 488/510 nm). Protect samples from light during all steps. Perform flow analysis within 1 h after staining.
Note 5: Use unstained cells suspended only in Assay Buffer as the negative control for instrument setup.
Note 6: Flow cytometry is highly sensitive; the working dye concentration may be lower than that used for fluorescence microscopy. Adjust the dilution factor of BODIPY 493/503 according to cell type and staining results.
(V) Detection by Fluorescence Microplate Reader
1.Cell seeding and treatment: Seed cells in black 96-well plates at 100–10,000 cells per well, typically 2000–5000 cells per well. Treat cells according to the experimental design.
2.Fixation (optional): Add 4% paraformaldehyde fixative and incubate at room temperature for 10–15 min.
Notes 1, 2, 3: Same as above.
3.Washing: For adherent cells, aspirate liquid and wash cells twice with PBS. For suspension cells, centrifuge at 250–1000 ×g for 5 min at room temperature, discard supernatant, and wash twice with PBS.
4.Staining: Add an appropriate volume of staining working solution, typically 100 μL per well for 96-well plates. Incubate at room temperature in the dark for 10–20 min. Optimal incubation time varies by cell type; start with 10 min and optimize based on staining results.
5.Detection: Wash cells twice with PBS after incubation. Read using a fluorescence microplate reader with appropriate wavelengths based on probe spectra (e.g., Ex/Em = 488/510 nm). Wavelengths other than the maximum excitation/emission pair may be used depending on instrument specifications.
Precautions
1.Upon first use, aliquot the kit and store at -20 °C to avoid repeated freeze‑thaw cycles. Seal aliquots according to single-use volumes.
2.Optimize BODIPY 493/503 working concentration, cell number, incubation temperature and duration for your cell type. The BODIPY 493/503 concentration can be adjusted within 0.2× ~ 2×.
3.BODIPY 493/503 is susceptible to photobleaching under laser irradiation. Use the minimum effective dye concentration, reduce microscope excitation power and shorten imaging time while maintaining sufficient fluorescence intensity.
4.For reliable results, analyze samples within 1 hour after staining.
5.All fluorescent dyes are prone to photobleaching; minimize light exposure to slow quenching.
6.Dispense Assay Buffer under aseptic conditions to prevent microbial contamination that impairs performance. Assay Buffer can be stored short‑term at 2–8 °C and is valid for one month.
7.This product is for research use only by qualified professionals. It shall not be used for clinical diagnosis or therapy, food or pharmaceutical applications, and shall not be stored in residential premises.
8.Wear a lab coat and disposable gloves for personal safety and protection.
规格
规格或纯度
BioReagent, 即用型, 适用于荧光分析, 无菌
英文名称
Fluorescent Lipid Droplet Assay Kit (BODIPY 493/503, Green)
应用
细胞染色
样品类型
细胞
检测方法
荧光法
定量&半定量
半定量
检测仪器
流式细胞仪,荧光显微镜,荧光酶标仪
检测波长
BODIPY 493/503：Ex/Em:493/503nm Hoechst 33342：Ex/Em:346/460nm
敏感性
Light-sensitive
Emission(nm)
460nm, 503nm
Excitation(nm)
Blue Laser(488nm), Ultraviolet Laser(350nm)
Ex/Em(nm)
Ex:493nm;Em:503nm,Ex:346nm;Em:460nm
储存与运输
储存条件
避光,-20°C储存
运输条件
超低温运输
稳定性与储存
Store at -20℃ long term (12 months). Store in the dark.
组分与储存
| 组分货号 | 组分名称 | 物理外观 | 100-1000T | 500-5000T | 储存 |
| F1527962A | BODIPY 493/503 (1000×) | 液体 | 0.1mL | 0.5mL | -20℃. 避光 |
| F1527962B | Hoechst 33342 (1000×) | 液体 | 0.1mL | 0.5mL | -20℃. 避光 |
| F1527962C | 检测缓冲液 | 液体 | 100mL | 500mL | -20℃. 避光 |
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
技术文档和文章
溶液计算器
需要帮助选择级别？
我们的级别选择指南涵盖了目录中所有的变体的纯度、稳定剂状态和应用适用性。
查看 BioReagent 级别指南 → 查看 Ready-to-use 级别指南 → 查看 for Fluorescence analysis 级别指南 → 查看 Sterile 级别指南 →产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
Copyright © 2023至今 上海阿拉丁生化科技股份有限公司 保留所有权利。
危险品化学品经营许可证（带存储）