Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe. / Harsløf, Mads; Müller, Christoph Felix; Rohrberg, Julie; Rekling, Jens C.

In: Journal of Neuroscience Methods, Vol. 253, 30.09.2015, p. 101-9.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Harsløf, M, Müller, CF, Rohrberg, J & Rekling, JC 2015, 'Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe', Journal of Neuroscience Methods, vol. 253, pp. 101-9. https://doi.org/10.1016/j.jneumeth.2015.06.005

APA

Harsløf, M., Müller, C. F., Rohrberg, J., & Rekling, J. C. (2015). Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe. Journal of Neuroscience Methods, 253, 101-9. https://doi.org/10.1016/j.jneumeth.2015.06.005

Vancouver

Harsløf M, Müller CF, Rohrberg J, Rekling JC. Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe. Journal of Neuroscience Methods. 2015 Sep 30;253:101-9. https://doi.org/10.1016/j.jneumeth.2015.06.005

Author

Harsløf, Mads ; Müller, Christoph Felix ; Rohrberg, Julie ; Rekling, Jens C. / Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe. In: Journal of Neuroscience Methods. 2015 ; Vol. 253. pp. 101-9.

Bibtex

@article{fb50930311cd4ef1a7cf9dacb87217ce,

title = "Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe",

abstract = "BACKGROUND: Biocytin has found numerous uses as a neuronal tracer, since it shows both antero- and retrograde transport in neuronal tracts. The main advantage of biocytin lies in the comprehensive intracellular distribution of the molecule, and in effective detection using avidin-based reactions. The main drawback is that biocytin cannot be visualized in live tissue.NEW METHOD: We demonstrate that TMR biocytin, a conjugate of biocytin and a rhodamine fluorophore, is an effective neuronal tracer in live tissue when applied by electroporation.RESULTS: The initial fiber transport velocity of TMR biocytin is high-5.4mm/h. TMR biocytin can be used in conjunction with AM calcium dyes to label neuronal somas from distances of several millimetres, and record calcium transients during the course of a few hours. Juxtacellular application of TMR biocytin leads to fast anterograde transport with labeling of local synapses within 10min. TMR biocytin is fixable, stable during methyl salicylate clearing, and can be visualized deep in nervous tissue.COMPARISON WITH EXISTING METHODS: Retrograde labeling with TMR biocytin enables long-range neuronal visualization and concurrent calcium imaging after only a few hours, which is substantially faster than other fluorescence-based tracers. The green emitting Atto 488 biotin is also taken up and transported retrogradely, but it is not compatible with standard green emitting calcium dyes.CONCLUSIONS: TMR biocytin is an attractive neuronal tracer. It labels neurons fast over long distances, and it can be used in conjunction with calcium dyes to report on neuronal activity in retrogradely labeled live neurons.",

author = "Mads Harsl{\o}f and M{\"u}ller, {Christoph Felix} and Julie Rohrberg and Rekling, {Jens C}",

year = "2015",

month = sep,

day = "30",

doi = "10.1016/j.jneumeth.2015.06.005",

language = "English",

volume = "253",

pages = "101--9",

journal = "Journal of Neuroscience Methods",

issn = "0165-0270",

publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Fast neuronal labeling in live tissue using a biocytin conjugated fluorescent probe

AU - Harsløf, Mads

AU - Müller, Christoph Felix

AU - Rohrberg, Julie

AU - Rekling, Jens C

PY - 2015/9/30

Y1 - 2015/9/30

N2 - BACKGROUND: Biocytin has found numerous uses as a neuronal tracer, since it shows both antero- and retrograde transport in neuronal tracts. The main advantage of biocytin lies in the comprehensive intracellular distribution of the molecule, and in effective detection using avidin-based reactions. The main drawback is that biocytin cannot be visualized in live tissue.NEW METHOD: We demonstrate that TMR biocytin, a conjugate of biocytin and a rhodamine fluorophore, is an effective neuronal tracer in live tissue when applied by electroporation.RESULTS: The initial fiber transport velocity of TMR biocytin is high-5.4mm/h. TMR biocytin can be used in conjunction with AM calcium dyes to label neuronal somas from distances of several millimetres, and record calcium transients during the course of a few hours. Juxtacellular application of TMR biocytin leads to fast anterograde transport with labeling of local synapses within 10min. TMR biocytin is fixable, stable during methyl salicylate clearing, and can be visualized deep in nervous tissue.COMPARISON WITH EXISTING METHODS: Retrograde labeling with TMR biocytin enables long-range neuronal visualization and concurrent calcium imaging after only a few hours, which is substantially faster than other fluorescence-based tracers. The green emitting Atto 488 biotin is also taken up and transported retrogradely, but it is not compatible with standard green emitting calcium dyes.CONCLUSIONS: TMR biocytin is an attractive neuronal tracer. It labels neurons fast over long distances, and it can be used in conjunction with calcium dyes to report on neuronal activity in retrogradely labeled live neurons.

AB - BACKGROUND: Biocytin has found numerous uses as a neuronal tracer, since it shows both antero- and retrograde transport in neuronal tracts. The main advantage of biocytin lies in the comprehensive intracellular distribution of the molecule, and in effective detection using avidin-based reactions. The main drawback is that biocytin cannot be visualized in live tissue.NEW METHOD: We demonstrate that TMR biocytin, a conjugate of biocytin and a rhodamine fluorophore, is an effective neuronal tracer in live tissue when applied by electroporation.RESULTS: The initial fiber transport velocity of TMR biocytin is high-5.4mm/h. TMR biocytin can be used in conjunction with AM calcium dyes to label neuronal somas from distances of several millimetres, and record calcium transients during the course of a few hours. Juxtacellular application of TMR biocytin leads to fast anterograde transport with labeling of local synapses within 10min. TMR biocytin is fixable, stable during methyl salicylate clearing, and can be visualized deep in nervous tissue.COMPARISON WITH EXISTING METHODS: Retrograde labeling with TMR biocytin enables long-range neuronal visualization and concurrent calcium imaging after only a few hours, which is substantially faster than other fluorescence-based tracers. The green emitting Atto 488 biotin is also taken up and transported retrogradely, but it is not compatible with standard green emitting calcium dyes.CONCLUSIONS: TMR biocytin is an attractive neuronal tracer. It labels neurons fast over long distances, and it can be used in conjunction with calcium dyes to report on neuronal activity in retrogradely labeled live neurons.

U2 - 10.1016/j.jneumeth.2015.06.005

DO - 10.1016/j.jneumeth.2015.06.005

M3 - Journal article

C2 - 26079494

VL - 253

SP - 101

EP - 109

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

ER -

ID: 152986876

Department of Neuroscience