aCell-Tracker Orange CMTMR 活細胞示蹤探針（橙色）

產品關鍵詞：

Cell-Tracker Green CMFDA；CellTracker™ Orange CMTMR；Cell movement 細胞運動；Cell location 細胞定位；Cell Tracker dye 細胞示蹤探針；CAS NO：323192-14-9；

訂購信息：

產品描述

Cell-Tracker熒光探針是用來監測細胞運動、定位、增殖、遷移、趨化和侵襲的優秀工具。

Cell-Tracker Orange CMTMR，英文全名：5-(and-6)-(((4-chloromethyl)benzoyl)amino)tetramethylrhodamine) (mixed isomers)，能自由穿透細胞膜進入細胞，在胞內轉化生成不具細胞膜滲透性的反應產物。該產物幾次傳代都能良好的保留在活細胞。在細胞群內，染料只會轉移到子代細胞，不會轉移到鄰近細胞。

Cell-Tracker Orange CMTMR特地設計使其至少72h（典型有3~6代）能展示熒光，此染料表現出理想的示蹤特征：穩定、工作濃度下無毒性、良好保留在細胞，且在生理pH下呈明亮熒光。另外，Cell-Tracker Orange CMTMR的激發和發射光譜（541/565nm）與綠色熒光蛋白GFP很好的分開，適用于多重標記（見附表1. Cell-Tracker熒光探針的光譜特征）。

產品特性

1）  CAS NO.：323192-14-9

2）  同義名：CellTracker™ Orange CMTMR;Xanthylium, 9-[2-carboxy-4(or 5)-[[4-(chloromethyl) benzoyl]amino]phenyl]-3,6-bis(dimethylamino)-, inner salt

3）  分子式：C32H28ClN3O4

4）  分子量：554.04 g/mol

5）  外觀：紫色固體

6）  溶解性：溶于DMSO

7）  化學結構式： 

保存與運輸方法

保存：-20℃避光干燥保存，至少2年有效。

運輸：冰袋運輸。

注意事項

1. 熒光染料都存在淬滅的問題，保存和操作過程中注意避光。

2. 避免使用含氨基和巰基的緩沖液。

3. 為了您的安全和健康，請穿實驗服并戴一次性手套操作。

應用示例

文獻1）McKee AS et al.Host DNA released in response to aluminum adjuvant enhances MHC class II-mediated antigen presentation and prolongs CD4 T-cell interactions with dendritic cells. Proc Natl Acad Sci U S A. 2013 Mar 19;110(12): E1122-31. PMID: 23447566

操作方法（多光子顯微鏡）：CD4 T cellswere isolated from OTII or B6 mice using the CD4 T-cell negative isolation kit andwere labeled with 20 μM CMTMR or 2 μM CFSE, washed three times, and injected i.v. into B6 or CD11c-YFP recipients. Twenty-four hours later, these recipient mice were immunized with 20 μg of AF647-labeled ova + 200 μg of Alhydrogel and 5 mg of either BSA or DNase. From 20–24 h following immunization, mice were killed and their draining popliteal LNs were surgically removed for imaging. 

文獻2）Nikitina EY et al.Cmbination ofγ‐irradiation and dendritic cell administration induces a potent antitumor response in tumor‐bearing mice: Approach to treatment of advanced stage cancer. Int J Cancer. 2001 Dec 15;94(6):825-33. PMID: 11745485

操作方法（組織內細胞分布評估）：DCs were labeled with 20 μM cell tracker orange (CMTMR) by incubation in RPMI 1640 at 37℃ for 30 min. Cells were then washed in PBS once, incubated for 30 min more and injected (107/mouse) intravenously (i.v.) or subcutaneously (s.c.) near the tumor site. Eighteen hours later lung, spleen, liver, superficial inguinal and popliteal LN as well as tumor tissue were taken out and snap‐frozen in tissue freezing media at?80°C. Frozen tissue sections were examined by fluorescence microscopy using a 540 nm filter. Labeled cells were counted in 10 fields at the total magnification ×200.

Fig. DC labeled with cell tracker CMTMR inside MethA sarcma. Unlabeled (I) or CMTMR‐labeled (II, III) DCs (107) were injected i.v. (II) or s.c. (III) into MethA sarcma‐bearing mice 3 hr after irradiation of the tumor with 10 Gy. Tumors were excised 18 hr later and cryostat sections were performed. Slides were analyzed by fluorescence microscopy using a 540‐nm filter. One of the representative fields with a total magnification of 200× is shown. 

使用方法

1．細胞準備

在合適的培養基內培養細胞。貼壁細胞可以在含蓋玻片的培養皿內爬片生長，裝入足量的生長培養基。

2．操作步驟

以下描述的是將染料加到培養細胞以及在熒光顯微鏡下成像的步驟。各種因素，比如將染料加載到細胞或組織，可能都需根據特定的細胞類型對某些條件做出修改。

探針的染色濃度需根據用途來調整。建議剛開展實驗需要測定至少1個10倍范圍內的濃度。一般來說，長期染色（≥3天）或使用快速分裂的細胞需5-25µM的染料。對于短期實驗（比如活力測定），使用低濃度染料（0.5-5µM）。為了維持正常的細胞形態和降低潛在的偽影，盡可能使用低濃度的染料。

2.1制備Cell-Tracker染色工作液

①開瓶前將產品從冰箱取出，放到室溫使其回溫至少20min。

②用高質量的無水DMSO溶解粉末使其濃度為10mM。例如，對1mgCell-Tracker Green CMFDA（Mw:464.86 g/mol）凍干粉，加入215µlDMSO充分溶解，即得到10mM母液。

③ 用無血清培養基稀釋母液到0.5-25µM的工作濃度。預熱染色工作液到37℃。

2.2懸浮細胞染色步驟

① 離心收集細胞，吸掉上清液。用預熱的Cell-Tracker染色工作液輕輕的重懸細胞。

② 在適合特定細胞類型的生長條件下孵育15-45min。

③ 離心細胞，吸掉Cell-Tracker染色工作液。

④ 加入選擇的培養基，將標記好的細胞分配到載玻片或到選擇的培養器皿內。

⑤根據附表1選擇合適激發和發射波長的濾片來進行成像檢測。

2.3貼壁細胞染色步驟

① 吸走培養基。

② 輕輕加入預熱的Cell-Tracker染色工作液。

③ 在適合特定細胞類型的生長條件下孵育15-45min。

④吸掉Cell-Tracker染色工作液。

⑤ 加入選擇的培養基。

⑥根據表1選擇合適激發和發射波長的濾片來進行成像檢測。

3．熒光顯微鏡觀察

Cell-Tracker熒光探針可用帶標準光學和圖像增強的各種落射熒光光學顯微鏡檢測。根據染料選擇合適的濾片。見附表1 Cell-Tracker熒光探針的光譜特征。

相關產品

— —Written/Edited by V. Shallan【版權歸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學和生物技術領域研究的試劑、儀器和實驗室消耗品與實驗服務工作，主要從事細胞生物學、植物學、分子生物學、免疫學、生物化學、蛋白組學。生物制藥與診斷試劑研發生產等領域。 本公司秉承“以人為本，以誠為信、合同守信"的經營理念。堅持"品質保障"的原則為廣大客戶提供良好產品。

引用文獻：

[1]  Sha, X., Dai, Y., Chong, L. et al. Pro-efferocytic macrophage membrane biomimetic nanoparticles for the synergistic treatment of atherosclerosis via copetition effect. J Nanobiotechnol 20, 506 (2022).

使用產品：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

使用產品：Cell-Tracker Red CMTPX and Cell-Tracker Green CMFDA

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

使用產品：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 cmplex 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.

使用產品：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.

使用產品：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)

使用產品：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.

使用產品：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.

使用產品：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.

使用產品：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).

使用產品：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.

使用產品：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

使用產品：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 Cmbined 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.

使用產品：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.

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