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Cell Explorer™ Live Cell Labeling Kit *Green Fluorescence*

Image of HeLa cells stained with Cell Explorer™ Live Cell Labeling Kit *Green Fluorescence* (Cat#22607) in a Costar black wall/clear bottom 96-well plate. Cells were stained with Calcein Green™ for 30 minutes. Images were aquired with fluorescence microscopy with FITC filter.
Image of HeLa cells stained with Cell Explorer™ Live Cell Labeling Kit *Green Fluorescence* (Cat#22607) in a Costar black wall/clear bottom 96-well plate. Cells were stained with Calcein Green™ for 30 minutes. Images were aquired with fluorescence microscopy with FITC filter.
Ordering information
Price ()
Catalog Number22607
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Excitation (nm)494
Emission (nm)514
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200

OverviewpdfSDSpdfProtocol


Excitation (nm)
494
Emission (nm)
514
Our Cell Explorer™ fluorescence imaging kits are a set of tools for labeling cells for fluorescence microscopic investigations of cellular functions. The effective labeling of cells provides a powerful method for studying cellular events in a spatial and temporal context. This particular kit is designed to uniformly label live cells in green fluorescence. The kit uses non-fluorescent Calcein Green™ that becomes strongly fluorescent upon entering into live cells. Calcein Green™ is a hydrophobic compound that easily permeates intact live cells. The hydrolysis of the non-fluorescent Calcein Green™ by intracellular esterases generates the strongly fluorescent hydrophilic Calcein Green™ fluorophore that is well-retained in the cell cytoplasm. Cells grown in black-walled plates can be stained and quantified in less than two hours. It can be readily adapted for high-throughput assays in a wide variety of fluorescence platforms such as microplate assays, immunocytochemistry and flow cytometry. It is useful for a variety of studies, including cell adhesion, chemotaxis, multidrug resistance, cell viability, apoptosis and cytotoxicity. The kit provides all the essential components with an optimized cell-labeling protocol.

Platform


Fluorescence microscope

ExcitationFITC filter
EmissionFITC filter
Recommended plateBlack wall/clear bottom

Components


Component A: Calcein Green™2 vials
Component B: HHBS (Hanks' buffer with 20 mM Hepes)1 bottle (100 mL)

Example protocol


AT A GLANCE

Protocol summary

  1. Prepare cells in growth medium
  2. Remove the medium
  3. Add Calcein Green™ working solution (100 µL/well for 96-well plates or 25 µL/well for 384-well plates)
  4. Incubate the cells at 37 oC for 30 - 60 minutes
  5. Wash the cells
  6. Examine the specimen under under fluorescence microscope with FITC filter (Ex/Em = 490/525 nm)

Important notes
Thaw all the components at room temperature before starting the experiment.

PREPARATION OF STOCK SOLUTION

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. Calcein Green™ stock solution:
Add 20 µL of DMSO into the vial of Calcein Green™ (Component A) and mix well to make Calcein Green™ stock solution. Note: 20 µL of Calcein Green™ stock solution is enough for 1 plate. For storage, seal tubes tightly. Note: Unused Calcein Green™ stock solution can be aliquoted and stored at < -20 oC for more than one month if the tubes are sealed tightly. Avoid repeated freeze-thaw cycles and protect from light.

PREPARATION OF WORKING SOLUTION

Add 20 µL of Calcein Green™ stock solution into 10 mL of HHBS (Component B) and mix well to make Calcein Green™ working solution. Note: Protect from light.  

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

  1. Remove the growth medium from the cell plates.

  2. Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Calcein Green™ working solution into the cell plate.

  3. Incubate the cells in a 37°C, 5% CO2 incubator for 30 to 60 minutes. 

  4. Wash the cells with HHBS (Component B), and add growth medium or HHBS back to the cells.

  5. Image the cells using a fluorescence microscope with FITC filter (Ex/Em = 490/525 nm).

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)494
Emission (nm)514

Citations


View all 3 citations: Citation Explorer
Autophagy proteins are not universally required for phagosome maturation
Authors: Cemma, Marija and Grinstein, Sergio and Brumell, John H
Journal: Autophagy (2016): 1440--1446
Differential detection of tumor cells using a combination of cell rolling, multivalent binding, and multiple antibodies
Authors: Myung, Ja Hye and Gajjar, Khyati A and Chen, Jihua and Molokie, Robert E and Hong, Seungpyo
Journal: Analytical chemistry (2014): 6088--6094
Versatile fabrication of nanoscale sol--gel bioactive glass particles for efficient bone tissue regeneration
Authors: Lei, Bo and Chen, Xiaofeng and Han, Xue and Zhou, Jiaan
Journal: Journal of Materials Chemistry (2012): 16906--16913

References


View all 26 references: Citation Explorer
Requirements, features, and performance of high content screening platforms
Authors: Gough AH, Johnston PA.
Journal: Methods Mol Biol (2007): 41
A pharmaceutical company user's perspective on the potential of high content screening in drug discovery
Authors: Hoffman AF, Garippa RJ.
Journal: Methods Mol Biol (2007): 19
Optimizing the integration of immunoreagents and fluorescent probes for multiplexed high content screening assays
Authors: Giuliano KA., undefined
Journal: Methods Mol Biol (2007): 189
Past, present, and future of high content screening and the field of cellomics
Authors: Taylor DL., undefined
Journal: Methods Mol Biol (2007): 3
High-content fluorescence-based screening for epigenetic modulators
Authors: Martinez ED, Dull AB, Beutler JA, Hager GL.
Journal: Methods Enzymol (2006): 21
Application of laser-scanning fluorescence microplate cytometry in high content screening
Authors: Bowen WP, Wylie PG.
Journal: Assay Drug Dev Technol (2006): 209
High-content screening of known G protein-coupled receptors by arrestin translocation
Authors: Hudson CC, Oakley RH, Sjaastad MD, Loomis CR.
Journal: Methods Enzymol (2006): 63
Evaluation of a high-content screening fluorescence-based assay analyzing the pharmacological modulation of lipid homeostasis in human macrophages
Authors: Werner T, Liebisch G, Gr and l M, Schmitz G.
Journal: Cytometry A (2006): 200
Automated high content screening for phosphoinositide 3 kinase inhibition using an AKT 1 redistribution assay
Authors: Wolff M, Haasen D, Merk S, Kroner M, Maier U, Bordel S, Wiedenmann J, Nienhaus GU, Valler M, Heilker R.
Journal: Comb Chem High Throughput Screen (2006): 339
High concordance of drug-induced human hepatotoxicity with in vitro cytotoxicity measured in a novel cell-based model using high content screening
Authors: O'Brien P J, Irwin W, Diaz D, Howard-Cofield E, Krejsa CM, Slaughter MR, Gao B, Kaludercic N, Angeline A, Bernardi P, Brain P, Hougham C.
Journal: Arch Toxicol (2006): 580

Application notes


Annexin V