long-term, irreversible membrane markers

Jorg Kirberg kirberg at nki.nl
Sun Aug 25 07:29:23 EST 1996

In article <kirberg-2408962228470001 at pcb72.nki.nl>, kirberg at nki.nl (Jorg
Kirberg) wrote:

> In article <4vktu5$1p2g at ds2.acs.ucalgary.ca>, kmuldrew at acs.ucalgary.ca
> (Ken Muldrew) wrote:
> > Has anyone ever heard of membrane dyes which are essentially
> > permanent? Perhaps with the name "zynaxis"? I'm trying to decide on a
> > method for marking graft cells as compared with host (xy vs. xx won't
> > work in this situation).
> > 
> > Ken Muldrew
> > kmuldrew at acs.ucalgary.ca
> Dear Ken,
> no idea about "zynaxis", but doesn't sigma sell some red fluorescent
> dye that binds to membranes ? There was even an add in the
> last science issue (Aug 16, opposide page 863).
> If I remember correctly, that is the same dye someone used in the
> institute I worked before. Try medline searching for
> Allen Young (Basel Institute for Immunology, Switzerland). He was
> quite pleased by the stuff he used. Fluorescence was long lasting as
> well.
> jorg
And here it is :
(Allen Young is now in Basel, apparently he published this before).
AU  - Young AJ
AU  - Hay JB
IN  - Trauma Research Program, Sunnybrook Health Science Centre, New York,
      Ontario, Canada.
TI  - Rapid turnover of the recirculating lymphocyte pool in vivo.
SO  - Int Immunol 1995 Oct;7(10):1607-15
MH  - Animal
MH  - Body Fluid Compartments
MH  - Cell Aging
MH  - Cell Movement
MH  - Comparative Study
MH  - Erythrocyte Aging
MH  - Female
MH  - Fluorescent Dyes/pk [Pharmacokinetics]
MH  - *Lymphocytes/cy [Cytology]
MH  - Organ Specificity
MH  - Sheep/ph [Physiology]
MH  - Support, Non-U.S. Gov't
MH  - Time Factors
AB  - Lymphocytes are unique among blood cells in their capacity to
      continually recirculate between blood and the tissues via the lymph.
      Previous estimates of lymphocyte lifespan in vivo and the turnover
      of the recirculating lymphocyte pool have been deduced from indirect
      labeling techniques. Using the fluorescent dye PKH-26, individual
      labeled cells have been tracked in sheep for periods > 2 months. By
      direct measurement their lifespan was calculated. This label was
      found to be stable in vivo, allowing long-term analysis of the
      characteristics of the recirculating lymphocyte pool. It is possible
      to calculate the rate of turnover of cells of the recirculating pool
      based on the rate at which labeled cells disappear from the
      lymphatic circulation. The recirculating lymphocyte pool was found
      to repopulate itself every 16.5 +/- 3.0 days. Using this label, it
      was estimated that recirculating lymphocytes divide on average once
      every 29.8 +/- 6.8 days. Labeled erythrocytes were also examined and
      found to have an average lifespan of 153 days, demonstrating no dye
      loss over the 2 month period of observation. These data suggest that
      the recirculating lymphocyte pool is a highly dynamic compartment,
      with a high rate of turnover and peripheral cell division in vivo.
      This is the first report of the direct measurement of the in vivo
      turnover of recirculating lymphocyte pools, and this method may now
      be used to further analyze the lifespan of individual lymphocyte
      subsets and the in vivo lifespan of other cell types in vivo.
EM  - 9605.  Entry Week: 96052.



Joerg Kirberg                              Tel. 0031 - 20 - 512 19 98
The Netherlands Cancer Institute           FAX  0031 - 20 - 512 20 11
Div. Molecular Genetics (H4)               E_mail kirberg at nki.nl
Plesmanlaan 121
NL - 1066 CX Amsterdam
The Netherlands

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