Calcein-AM Cell Analysis Live Cell viability Stain 1mg Cellstain® Dojindo Laboratories Wako

Cellstain® Calcein-AM 1mg produced by Dojindo Laboratories and distributed in association with FUJIFILM Wako Chemicals Europe GmbH

The Cellstain® range, produced by Dojindo Laboratories is intended for research use to facilitate cell analysis. Cellstain® Calcein-AM is a live cell stain. Calcein-AM readily passes through the cell membrane of viable cells because of its enhanced hydrophobicity compared to Calcein. After Calcein-AM permeates into the cytoplasm, it is hydrolyzed by esterases to Calcein, which remains inside the cell (Fig. 1). Among other reagents, including BCECF-AM and Carboxy-fluorescein diacetate, Calcein-AM is the most suitable fluorescent probe for staining viable cells because of its low cytotoxicity. Calcein does not inhibit any cellular functions such as proliferation or chemotaxis of lymophocyte. In addition, viability assays using Calcein are reliable and correlate well with the standard ⁵¹Cr-release assay. The excitation and emission wavelengths of calcein are 490 nm and 515 nm, respectively

Staining Procedure
1. Prepare 1 mM Calcein-AM solution with DMSO and dilute to prepare 1-50 µM Calcein-AM solution with PBS.^a)
2. Add Calcein-AM solution with 1/10 of the volume of cell culture medium to the cell culture. ^b)
3. Incubate the cell at 37°C for 15-30 min.
4. Wash cells twice with PBS or an appropriate buffer.
5. Observe the cells under a fluorescence microscope with 490 nm excitation and 515 nm emission filters.
a) If the Calcein-AM has difficulty loading into cells, use a detergent such as Pluronic F127.
b) Or you may replace the culture medium with 1/10 concentration of Calcein-AM buffer solution.
Cellstain® Calcein-AM 1mg
Also known as 3',6'-Di(O-acetyl)-4',5'-bis[N,N-bis(carboxymethyl)aminomethyl]fluorescein tetraacetoxymethyl ester
Dojindo Laboratories: C326-10
Pack Size: 1mg
Molecular Formula:C₄₈H₄₆N₂O₂₃
MW: 994.86
CAS No: 148504-34-1
Store -20°C, protect from light
Ships with ice packs

Reference

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2. S. J. Morris, Real-time Multi-wavelength Fluorescence Imaging of Living Cells. BioTechniques. 1990;8:296-308.
3. S. A. Weston, et al., New Fluorescent Dyes for Lymphocyte Migration Studies Analysis by Flow Cytometry and Fluorescent Microscopy. J Immunol Methods. 1990;133:87-97.
4. S. A. Weston, et al., Calcein: a Novel Marker for Lymphocytes Which Enter Lymph Nodes. Cytometry. 1992;13:739-749.
5. X. M. Wang, et al., A New Microcellular Cytotoxicity Test Based on Calcein AM Release. Hum Immunol. 1993;37:264-270.
6. N. G. Papadopoulos, et al., An Improved Fluorescence Assay for the Determination of Lymphocyte-Mediated Cytotoxicity Using Flow Cytometry. J Immunol Methods. 1994;177:101-111.
7. H. Ohata, et al., Confocal Imaging Analysis of ATP-Induced Ca2+ Response in Individual Endothelial Cells of the Artery in Situ. Am J Physiol. 1997;272:C1980-C1987.