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

產(chǎn)品關(guān)鍵詞：

Cell-Tracker Green CMFDA；CellTracker™ Orange CMTMR；Cell movement 細(xì)胞運(yùn)動(dòng)；Cell location 細(xì)胞定位；Cell Tracker dye 細(xì)胞示蹤探針；CAS NO：323192-14-9；

訂購(gòu)信息：

產(chǎn)品描述

Cell-Tracker熒光探針是用來(lái)監(jiān)測(cè)細(xì)胞運(yùn)動(dòng)、定位、增殖、遷移、趨化和侵襲的優(yōu)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(xì)胞膜進(jìn)入細(xì)胞，在胞內(nèi)轉(zhuǎn)化生成不具細(xì)胞膜滲透性的反應(yīng)產(chǎn)物。該產(chǎn)物幾次傳代都能良好的保留在活細(xì)胞。在細(xì)胞群內(nèi)，染料只會(huì)轉(zhuǎn)移到子代細(xì)胞，不會(huì)轉(zhuǎn)移到鄰近細(xì)胞。

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

產(chǎn)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(xué)結(jié)構(gòu)式： 

保存與運(yùn)輸方法

保存：-20℃避光干燥保存，至少2年有效。

運(yùn)輸：冰袋運(yùn)輸。

注意事項(xiàng)

1. 熒光染料都存在淬滅的問(wèn)題，保存和操作過(guò)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(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。

應(yīng)用示例

文獻(xiàn)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(xiàn)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(nèi)細(xì)胞分布評(píng)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(xì)胞準(zhǔn)備

在合適的培養(yǎng)基內(nèi)培養(yǎng)細(xì)胞。貼壁細(xì)胞可以在含蓋玻片的培養(yǎng)皿內(nèi)爬片生長(zhǎng)，裝入足量的生長(zhǎng)培養(yǎng)基。

2．操作步驟

以下描述的是將染料加到培養(yǎng)細(xì)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(xì)胞或組織，可能都需根據(jù)特定的細(xì)胞類型對(duì)某些條件做出修改。

探針的最佳染色濃度需根據(jù)用途來(lái)調(diào)整。建議剛開展實(shí)驗(yàn)需要測(cè)定至少1個(gè)10倍范圍內(nèi)的濃度。一般來(lái)說(shuō)，長(zhǎng)期染色（≥3天）或使用快速分裂的細(xì)胞需5-25µM的染料。對(duì)于短期實(shí)驗(yàn)（比如活力測(cè)定），使用低濃度染料（0.5-5µM）。為了維持正常的細(xì)胞形態(tài)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(chǎn)品從冰箱取出，放到室溫使其回溫至少20min。

②用高質(zhì)量的無(wú)水DMSO溶解粉末使其濃度為10mM。例如，對(duì)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(wú)血清培養(yǎng)基稀釋母液到0.5-25µM的工作濃度。預(yù)熱染色工作液到37℃。

2.2懸浮細(xì)胞染色步驟

① 離心收集細(xì)胞，吸掉上清液。用預(yù)熱的Cell-Tracker染色工作液輕輕的重懸細(xì)胞。

② 在適合特定細(xì)胞類型的生長(zhǎng)條件下孵育15-45min。

③ 離心細(xì)胞，吸掉Cell-Tracker染色工作液。

④ 加入選擇的培養(yǎng)基，將標(biāo)記好的細(xì)胞分配到載玻片或到選擇的培養(yǎng)器皿內(nèi)。

⑤根據(jù)附表1選擇合適激發(fā)和發(fā)射波長(zhǎng)的濾片來(lái)進(jìn)行成像檢測(cè)。

2.3貼壁細(xì)胞染色步驟

① 吸走培養(yǎng)基。

② 輕輕加入預(yù)熱的Cell-Tracker染色工作液。

③ 在適合特定細(xì)胞類型的生長(zhǎng)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(yǎng)基。

⑥根據(jù)表1選擇合適激發(fā)和發(fā)射波長(zhǎng)的濾片來(lái)進(jìn)行成像檢測(cè)。

3．熒光顯微鏡觀察

Cell-Tracker熒光探針可用帶標(biāo)準(zhǔn)光學(xué)和圖像增強(qiáng)的各種落射熒光光學(xué)顯微鏡檢測(cè)。根據(jù)染料選擇合適的濾片。見附表1 Cell-Tracker熒光探針的光譜特征。

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— —Written/Edited by V. Shallan【版權(quán)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(xué)和生物技術(shù)領(lǐng)域研究的試劑、儀器和實(shí)驗(yàn)室消耗品與實(shí)驗(yàn)服務(wù)工作，主要從事細(xì)胞生物學(xué)、植物學(xué)、分子生物學(xué)、免疫學(xué)、生物化學(xué)、蛋白組學(xué)。生物制藥與診斷試劑研發(fā)生產(chǎn)等領(lǐng)域。 本公司秉承“以人為本，以誠(chéng)為信、合同守信"的經(jīng)營(yíng)理念。堅(jiān)持"品質(zhì)保障"的原則為廣大客戶提供優(yōu)質(zhì)產(chǎn)品。

引用文獻(xiàn)：

[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).

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[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

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[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

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[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(chǎn)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(chǎn)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(chǎn)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(chǎn)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(chǎn)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.

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[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).

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[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(chǎn)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

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[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.

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[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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