I am doing retrograde neuronal labeling using the NeuroTrace DiI Paste. I apply a small glob onto the neuron of interest, but it is not adhering very well. What can I do?
This paste, which has DiI C18 uniformly dispersed within a proprietary hydrophobic carrier, is commonly used to dab onto larger areas than smaller areas commonly used with crystals. Any moisture on the area the paste is applied may be the cause for lack of adherence. Apply the paste using a small flat-head spatula or toothpick and keep it in place a bit longer to ensure contact. The area may also be covered with parafilm to ensure contact as well.
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How long does it take for lipophlic tracers to transport along the membrane? How much faster are the FAST lipophilic dyes?
The transport is fairly slow, around 6 mm/day in live tissue and slower in fixed tissue, so diffusion of lipophilic carbocyanine tracers from the point of their application to the terminus of a neuron can take several days to weeks The FAST DiO and DiI analogs (which have unsaturated alkyl tails) can improve transport rate by around 50%.
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Which form of the lipophilic tracers (DiO, DiI, DiD, etc) should I use?
Select the dye that is compatible with your available excitation source(s) and emission filter set/channels. The solid, paste and crystal forms can be applied directly to neurons in tissues. For labeling cells in culture or microinjection, the lipophilic dyes in solution or solid form can be used.
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What products do you have for neuronal tracing?
Please check out this web page (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing.html) for details.
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