Green CMFDA (Synonyms: 5-Chloromethylfluorescein diacetate)

Cell lines

platelets

Preparation Method

Samples of resting platelets and platelets activated by 100 µM ADP were adjusted to the platelet concentration of 3.5~3.8 × 108/L with autologous plasma, and incubated for 30 minutes at 37°C in 10 µM CMFDA. After incubation, the cytosolic esterase-induced fluorescence of platelets was measured by laser confocal microscopy and also photograph was taken.

Reaction Conditions

for 30 minutes at 37°C in 10 µM

Applications

The observation with laser confocal microscopy on CMFDA-stained platelets shows that nearly all the resting platelets release bright green fluorescence, whereas the ADP activated platelets release no or little fluorescence.

Animal models

NOD/SCID or NOD/SCID/β2Mnull mice

Preparation Method

To determine the optimal time of BMDC injection in relation to bleo injury, 5 × 106 CMFDA-labeled BMDC were washed, resuspended in Hanks' buffer with heparin (50 U/ml), and administered via tail vein injection to NOD/SCID or NOD/SCID/β2Mnull mice 1, 2, 3, or 4 days post-bleo.

Dosage form

5 × 10⁶ ;1, 2, 3, or 4 days

Applications

There was a 10- to 100-fold increase in the percentage of CMFDA+ cells seen in NOD/SCID/β2Mnull mice compared with NOD/SCID mice, with ∼0.04% positive cells seen when cells were infused at day 4 and mice were killed at day 7 after bleo.

References:

[1] Wang J, et al. Correlation between the In Vitro Functionality of Stored Platelets and the Cytosolic Esterase-Induced Fluorescence Intensity with CMFDA. PLoS One. 2015 Sep 21;10(9):e0138509.
[2] Liebler JM, et al. Retention of human bone marrow-derived cells in murine lungs following bleomycin-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 2008 Aug;295(2):L285-92.