DiD perchlorate is a far-red fluorescent lipophilic cyanine dye. DiD perchlorate can rapidly and stably integrate into the phospholipid cell membrane. DiD perchlorate is used to cells tracking^[1][2][3].

Two weeks after injection of stained cells, a single, bright, DiD perchlorate (DiD)-positive cells located between 15 to 40 µm from the endosteum is found. Results reveal a progressive appearance of cell clusters of decreased dye intensity, consistent with the partitioning of DiD perchlorate label on cell division^[3].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

959.90

C₆₁H₉₉ClN₂O₄

127274-91-3

Room temperature in continental US; may vary elsewhere.

4deg;C, protect from light, stored under nitrogen

*该产品在溶液状态不稳定，建议您现用现配，即刻使用。

In Vitro:;

DMSO : 25 mg/mL (26.04 mM; ultrasonic and warming and heat to 60°C)

*

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；该产品在溶液状态不稳定，建议您现用现配，即刻使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂：;10% DMSO ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

  Solubility: ≥ 1.67 mg/mL (1.74 mM); Clear solution

  此方案可获得 ≥ 1.67 mg/mL (1.74 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 16.7 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂：;10% DMSO ;; 90% (20% SBE-β-CD in saline)

  Solubility: ≥ 1.67 mg/mL (1.74 mM); Clear solution

  此方案可获得 ≥ 1.67 mg/mL (1.74 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 16.7 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂：;10% DMSO ;; 90% corn oil

  Solubility: ≥ 1.67 mg/mL (1.74 mM); Clear solution

  此方案可获得 ≥ 1.67 mg/mL (1.74 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 16.7 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Kenji Yumoto, et al. A novel method for monitoring tumor dormancy using fluorescent dye DiD. Cytometry A. 2014 Jun; 85(6): 548–555.

  [2]. Meng Li, et al. In Vivo Tracking of Human Adipose-derived Mesenchymal Stem Cells in a Rat Knee Osteoarthritis Model with Fluorescent Lipophilic Membrane Dye. J Vis Exp. 2017; (128): 56273.

  [3]. Lo Celso C, et al. Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche.

1 to 5×10⁵ cells per mL HSPCs are stained with 5 μM DiD perchlorate (DiD) or 7.5 μM DiI in PBS without serum for 10 min at 37°C, washed once in PBS and immediately injected into the tail vein of recipient mice. Unless stated differently, each imaged CD45.2 mouse receive 8,000 to 15,000 labelled CD45.1 HSPCs together with 3 to 5×10⁵ CD45.2 supportive whole bone-marrow mononuclear cells to ensure survival^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Kenji Yumoto, et al. A novel method for monitoring tumor dormancy using fluorescent dye DiD. Cytometry A. 2014 Jun; 85(6): 548–555.

  [2]. Meng Li, et al. In Vivo Tracking of Human Adipose-derived Mesenchymal Stem Cells in a Rat Knee Osteoarthritis Model with Fluorescent Lipophilic Membrane Dye. J Vis Exp. 2017; (128): 56273.

  [3]. Lo Celso C, et al. Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche.