Dear Flowers Always some problems (probably for many of you). For some time we have tried to label human monocytes (cryopreserved and thawed) with Molecular Probes Vybrant Cell-Labeling Solution. Our major problem is aggregation of the cells. We have followed the protocol carefully, using cells as recomended of 1 x 106 /mL, without serum, and adding 5 mL of the Cell-Labeling Solution per mL of cell suspension, incubating for varying times (30 minutes 37oC, spinning the cells and washing a couple of times. Generally the cells have been aggregating or adhering to our plastic tubes (Falcon Polypropylene tubes 352070). Our intention has been to study cell migration in TransWell systems using this labeling technique. All suggestions to circumvent aggregation is most welcome. Thanks! Peter Kierulf Peter Kierulf,MD PhD. Professor I Clinical Chemistry Dep. Ulleval University Hospital N-0407 OSLO, NORWAY Telephone: +47 22 11 94 86 Telefax: +47 22 11 81 89 _-----------------------------------Mobil telphone: 922.85111 E-mail (RFC Formate): peter.kierulf@ioks.uio.no E-mai. (SA Formate): /G=peter/S=kierulf/OU=ioks/O=uio/PRMD=uninett/C=no
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