CFSE (5(6)-Carboxyfluorescein diacetate succinimidyl ester) is an intracellular fluorescent dye which can track proliferating cells. Covalently bound CFSE is divided equally between daughter cells, allowing discrimination of successive rounds of cell division^[1][2].

CFSE is a fluorescent dye which can track the cell division^[1]. To examine the ability of immune cells to migrate into live slices, a single-cell suspension of autologous splenocytes or PBMC is isolated, labeling them with CFSE, and adding them to the top of the slices on day 2 of culture. After 6 additional days of culture, the CFSE-labeling immune cells (green) are found to have migrated throughout the slices. No positive cells are found in the slices without the addition of CFSE-labeling immune cells^[2].

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

557.46

C₅₈H₃₈N₂O₂₂

150347-59-4

5(6)-羧基二乙酸荧光素 N-琥珀酰亚胺酯

Room temperature in continental US; may vary elsewhere.

-20deg;C, sealed storage, away from moisture and light

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture and light)

In Vitro:;

DMSO : 50 mg/mL (89.69 mM; Need ultrasonic)

*

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；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

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: ≥ 2.08 mg/mL (3.73 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (3.73 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 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% corn oil

  Solubility: ≥ 2.08 mg/mL (3.73 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (3.73 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 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. A Bruce Lyons, et al. Flow cytometric analysis of cell division by dilution of CFSE and related dyes. Curr Protoc Cytom. 2013;Chapter 9:Unit9.11.

  [2]. Xiuyun Jiang et al. Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment. Oncoimmunology. 2017; 6(7): e1333210.

Single-cell suspensions of splenocytes (or PBMC) are stained with 1 µM CFSE in PBS for 9 min at 37°C, combined with 20 mL of 10% FBS RPMI-1640 medium at RT for 2 min, centrifuged, washed, and counted. 2×10⁶ CFSE-labeling splenocytes are added to the top of the slices in 24-well membrane culture insert^[2].

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

• [1]. A Bruce Lyons, et al. Flow cytometric analysis of cell division by dilution of CFSE and related dyes. Curr Protoc Cytom. 2013;Chapter 9:Unit9.11.

  [2]. Xiuyun Jiang et al. Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment. Oncoimmunology. 2017; 6(7): e1333210.