Cell-Tracker Green CMFDA  活细胞示踪探针（绿色）

重要提醒（购买或初次使用前）请务必查阅：

产品关键词：

Cell-Tracker Green CMFDA；Cell-Tracker Red CMTPX；Cell movement 细胞运动；Cell location 细胞定位；Cell Tracker dye 细胞示踪探针；CAS NO：136832-63-8；

订购信息：

产品描述

Cell-Tracker荧光探针是用来监测细胞运动、定位、增殖、迁移、趋化和侵袭的优秀工具。

Cell-Tracker Green CMFDA，英文全名：5-chloromethylfluorescein diacetate，能自由穿透细胞膜进入细胞，在胞内转化生成不具细胞膜渗透性的反应产物。该产物几次传代都能良好的保留在活细胞。在细胞群内，染料只会转移到子代细胞，不会转移到邻近细胞。

Cell-Tracker Red CMTPX特地设计使其至少72h（典型有3~6代）能展示荧光，此染料表现出理想的示踪特征：稳定、工作浓度下无毒性、良好保留在细胞，且在生理pH下呈明亮荧光。另外，Cell-Tracker Red CMTPX的激发和发射光谱（492/517nm）与红色荧光蛋白RFP很好的分开，适用于多重标记（见附表1. Cell-Tracker荧光探针的光谱特征）。

产品特性

1）  CAS NO.：136832-63-8

2）  同义名：CellTracker™ Green CMFDA; Spiro(isobenzofuran-1(3H),9'-(9H)xanthen)-3-one, 3',6'-bis(acetyloxy)-5-(chloromethyl)-;

3）  分子式：C25H17ClO7

4）  分子量：464.86 g/mol

5）  外观：无色固体

6）  溶解性：溶于DMSO

7）  化学结构式： 

保存与运输方法

保存：-20℃避光干燥保存，至少2年有效。

运输：冰袋运输。

注意事项

1. 荧光染料都存在淬灭的问题，保存和操作过程中注意避光。

2. 避免使用含氨基和巯基的缓冲液。

3. 为了您的安全和健康，请穿实验服并戴一次性手套操作。

应用示例

文献1）Nasir NAM et al.Fluorescent cell tracer dye permits real‐time assessment of re‐epithelialization in a serum‐free ex vivo human skin wound assay. Wound Repair Regen. 2019 Jan;27(1):126-133.

Method (Cell tracker dye uptake within human ex vivo wounds in situ)：Four microliters of 25 μM CMFDA (or CMTPX) was added to the wound surface for 30 minutes, then washed dropwise with phosphate buffered saline.For cell viability studies, 1 μg/mL of propidium iodide (PI) was cotreated alongside CMFDA. Planimetric imaging was conducted using a Leica M205 FA upright Stereomicroscope using a 5x/0.50 pLANapo LWD objective at the equivalent of 64x magnification and captured using a DFC 565FX camera through LAS AF v3.1.0.8587 software. Specific band‐pass filter set for green fluorescent protein was used.

Fig. Epithelial repair of human ex vivo wounds progresses via an extending shield mechanism. Human ex vivo wounds were cultured in the presence of the fluorescent dyes. CMFDA and PI cotreatment at time of injury (D0) identifies live (CMFDA; green) and dead (PI; red) cells around the injury site (A), and these wounds traced to day 1 indicate PI‐labeled cells are no longer present (B).

文献2）Frazer J. Bye et al. Development of bilayer and trilayer nanofibrous/microfibrous scaffolds for regenerative medicine.Biomater. Sci., 2013, 1, 942-951.

Method (Cell migration into scaffolds at 7 days)：CellTracker red (CMTPX) or green (CMFDA) were applied to the hESMPs and fibroblasts respectively, prior to seeding. Cells were washed 3 times with 5 ml PBS then 10 ml of serum free cell-appropriate medium containing CellTracker (10 mM) was added and the cells incubated for 45 minutes at 37 °C. After incubation, the cells were washed 3 times with 5 ml PBS following which they were seeded onto scaffolds.Scaffolds could then be imaged in an Axon ImageExpress microscope (Molecular Devices, Sunnyvale, USA) at 570 nm λex–620 nm λem (CellTracker red) and 480 nm λex–533 nm λem (CellTracker green).

Fig. Co-culture of CellTracker™ labelled fibroblasts (green) and hESMPs (red) on bilayer membranes of either PHBV–PLA or PHBV–PCL. hESMPs were seeded on day 1 (red) onto the PHBV face of each bilayer. Fibroblasts were seeded on either the PLA or PCL face of the bilayer on day 2 (green) and then cultured for a further 7 days. In A fibroblasts (green) are confined to the PLA face after 7 days with no sign of hESMPs (red). On the opposite face (B, PHBV), hESMPs (red) are also present once again with no fibroblasts.

使用方法

1．细胞准备

在合适的培养基内培养细胞。贴壁细胞可以在含盖玻片的培养皿内爬片生长，装入足量的生长培养基。

2．操作步骤

以下描述的是将染料加到培养细胞以及在荧光显微镜下成像的步骤。各种因素，比如将染料加载到细胞或组织，可能都需根据特定的细胞类型对某些条件做出修改。

探针的最佳染色浓度需根据用途来调整。建议刚开展实验需要测定至少1个10倍范围内的浓度。一般来说，长期染色（≥3天）或使用快速分裂的细胞需5-25µM的染料。对于短期实验（比如活力测定），使用低浓度染料（0.5-5µM）。为了维持正常的细胞形态和降低潜在的伪影，尽可能使用低浓度的染料。

2.1制备Cell-Tracker染色工作液

①开瓶前将产品从冰箱取出，放到室温使其回温至少20min。

②用高质量的无水DMSO溶解粉末使其浓度为10mM。例如，对1mgCell-Tracker Green CMFDA（Mw:464.86g/mol）冻干粉，加入215µlDMSO充分溶解，即得到10mM母液。

③ 用无血清培养基稀释母液到0.5-25µM的工作浓度。预热染色工作液到37℃。

2.2悬浮细胞染色步骤

① 离心收集细胞，吸掉上清液。用预热的Cell-Tracker染色工作液轻轻的重悬细胞。

② 在适合特定细胞类型的生长条件下孵育15-45min。

③ 离心细胞，吸掉Cell-Tracker染色工作液。

④ 加入选择的培养基，将标记好的细胞分配到载玻片或到选择的培养器皿内。

⑤根据附表1选择合适激发和发射波长的滤片来进行成像检测。

2.3贴壁细胞染色步骤

① 吸走培养基。

② 轻轻加入预热的Cell-Tracker染色工作液。

③ 在适合特定细胞类型的生长条件下孵育15-45min。

④吸掉Cell-Tracker染色工作液。

⑤ 加入选择的培养基。

⑥根据表1选择合适激发和发射波长的滤片来进行成像检测。

 3．荧光显微镜观察

Cell-Tracker荧光探针可用带标准光学和图像增强的各种落射荧光光学显微镜检测。根据染料选择合适的滤片。见附表1 Cell-Tracker荧光探针的光谱特征。

相关产品

— —Written/Edited by V. Shallan【版权归MKBio懋康所有】

上海懋康生物科技有限公司是一家涉足于生命科学和生物技术领域研究的试剂、仪器和实验室消耗品与实验服务工作，主要从事细胞生物学、植物学、分子生物学、免疫学、生物化学、蛋白组学。生物制药与诊断试剂研发生产等领域。 本公司秉承“以人为本，以诚为信、合同守信”的经营理念。坚持"品质保障"的原则为广大客户提供优质产品。

引用文献：

[1]  Sha, X., Dai, Y., Chong, L. et al. Pro-efferocytic macrophage membrane biomimetic nanoparticles for the synergistic treatment of atherosclerosis via competition effect. J Nanobiotechnol 20, 506 (2022). https://doi.org/10.1186/s12951-022-01720-2

使用产品：Cell-Tracker Red CMTPX and Cell-Tracker Green CMFDA

[2] Pinghua Ling et al. DNA logic gate for cancer cells identification, detection, and modulation of cell–cell interactions. Microchemical Journal. Volume 207, December 2024, 111840

使用产品：Cell-Tracker Red CMTPX and Cell-Tracker Green CMFDA

[3] Qingqing Fang et al. Design of high-avidity multivalent ligand structures that target cells with high ligand economy.  Chemical Communications. Volume 58, Issue 70, 2022, Pages 9802-9805

使用产品：Cell-Tracker Red CMTPX

[4] Song J, Zhang H, Wang D, Wang J, Zhou J, Zhang Z, Wang J, Hu Y, Xu Q, Xie C, Lu W, Liu M. Hydrogel loading functionalized PAMAM/shRNA complex for postsurgical glioblastoma treatment. J Control Release. 2021 Oct 10;338:583-592. doi: 10.1016/j.jconrel.2021.08.052. Epub 2021 Sep 2. PMID: 34481020.

使用产品：Cell-Tracker Red CMTPX and Cell-Tracker Green CMFDA 

[5] Jia D, Lu Y, Lv M, Wang F, Lu X, Zhu W, Wei J, Guo W, Liu R, Li G, Wang R, Li J, Yuan F. Targeted co-delivery of resiquimod and a SIRPα variant by liposomes to activate macrophage immune responses for tumor immunotherapy. J Control Release. 2023 Aug;360:858-871. doi: 10.1016/j.jconrel.2023.07.030. Epub 2023 Jul 26. PMID: 37473808.

使用产品：Cell-Tracker Red CMTPX

[6] Xu, G., An, B., Wang, R. et al. RBM10 deficiency promotes brain metastasis by modulating sphingolipid metabolism in a BBB model of EGFR mutant lung adenocarcinoma. J Exp Clin Cancer Res 44, 95 (2025). https://doi.org/10.1186/s13046-025-03347-1

使用产品：Cell-Tracker Green CMFDA 

[7] Zhang Z, Luo Y, Ma Y, Zhou Y, Zhu D, Shen W, Liu J. Photocatalytic manipulation of Ca2+ signaling for regulating cellular and animal behaviors via MOF-enabled H2O2 generation. Sci Adv. 2024 Apr 19;10(16):eadl0263. doi: 10.1126/sciadv.adl0263. Epub 2024 Apr 19. PMID: 38640246; PMCID: PMC11029810.

使用产品：Cell-Tracker Red CMTPX

[8] Tang C, Wang H, Guo L, Zou C, Hu J, Zhang H, Zhou W, Yang G. CpG-Conjugated Silver Nanoparticles as a Multifunctional Nanomedicine to Promote Macrophage Efferocytosis and Repolarization for Atherosclerosis Therapy. ACS Appl Mater Interfaces. 2023 Nov 1. doi: 10.1021/acsami.3c11227. Epub ahead of print. PMID: 37910772.

使用产品：Cell-Tracker Green CMFDA 

[9] Qiao Y, Wang L, Xu W, Yang P, Tang C, Song D, Ling P, Gao F. Reversible Modulation of Cell-Cell Interactions Using Electrochemistry. ACS Appl Mater Interfaces. 2024 Aug 21;16(33):43341-43349. doi: 10.1021/acsami.4c08619. Epub 2024 Aug 5. PMID: 39103300.

使用产品：Cell-Tracker Red CMTPX and Cell-Tracker Green CMFDA 

[10] Zhao, H., Liu, R., Wang, L. et al. Artificial Macrophage with Hierarchical Nanostructure for Biomimetic Reconstruction of Antitumor Immunity. Nano-Micro Lett. 15, 216 (2023). https://doi.org/10.1007/s40820-023-01193-4

使用产品：Cell-Tracker Green CMFDA and Cell-Tracker Blue CMAC

[11] Wang Y, Sun Y, Zhang X, Wang S, Huang X, Xu K, Liu Y, Huang Y, Xu J, Wei X, Cheng H, Pan L, Wang J, Gu Z. A Granzyme B-Cleavable T Cell-Targeted Bispecific Cell Vesicle Connector for Reversing New-Onset Type 1 Diabetes. J Am Chem Soc. 2025 Feb 5;147(5):4167-4179. doi: 10.1021/jacs.4c13644. Epub 2025 Jan 27. PMID: 39869523.

使用产品：Cell-Tracker Deep Red

 [12] Sulei Zhang, Jun Sun, Shuaihang Guo, Yichen Wang, Yuheng Zhang, Jiao Lei, Xiaoli Liu, Hong Chen, Balancing functions of antifouling, nitric oxide release and vascular cell selectivity for enhanced endothelialization of assembled multilayers, Regenerative Biomaterials, Volume 11, 2024, rbae096, https://doi.org/10.1093/rb/rbae096

使用产品：Cell-Tracker Orange CMTMR and Cell-Tracker Green CMFDA

[13] Chen P, Han Y, Wang L, Zheng Y, Zhu Z, Zhao Y, Zhang M, Chen X, Wang X, Sun C. Spatially Resolved Metabolomics Combined with the 3D Tumor-Immune Cell Coculture Spheroid Highlights Metabolic Alterations during Antitumor Immune Response. Anal Chem. 2023 Oct 17;95(41):15153-15161. doi: 10.1021/acs.analchem.2c05734. Epub 2023 Oct 6. PMID: 37800909.

使用产品：Cell-Tracker Red CMTPX

[14] Li D, Li Q, Xu T, Guo X, Tang H, Wang W, Zhang W, Zhang Y. Pro-vasculogenic Fibers by PDA-Mediated Surface Functionalization Using Cell-Free Fat Extract (CEFFE). Biomacromolecules. 2024 Mar 11;25(3):1550-1562. doi: 10.1021/acs.biomac.3c01124. Epub 2024 Feb 27. PMID: 38411008.

使用产品：Cell-Tracker Green CMFDA