Are there opensource alternatives to Neurolucida for dendritic spines analysis ?

Kindly give suggestions regarding calculation of dendritic spine density .I have a doubt that whether i have to calculate the spine numbers directly under microscope or from acquired images . To find the percentage of immature and mature spines,do we have any formulas to be used like preference index?. how many neurons i should use to calculate the spine numbers. I could not find exact way.

Hi everybody,

I have two experimental groups (vehicle- vs drug-treated) and two time points. I plotted correlated activity of different brain regions in each group, at each time point. Now, I would like to determine whether the correlated brain activity statistically differs between the two groups and two time points (so 2x2 design, but no matching or pairing across time). So far, I could not find a method to compare the strength of the correlations between two (or more) groups. Does anyone have experience in this?

Many thanks!

Best, Kübra

I am looking for methods used for the visualisation of dendritic spines that can be further used for analysis. I recently came across papers which use DiI dye in finely powdered form to be applied on tissue sections for the same purpose. I would like to ask if it's possible that DiI dye can be dissolved in PBS and then can be applied to tissue sections. This might help in uniform labelling in the entire area of the section. If this does not work, please do tell me your suggestions.

Hi everyone,

I want to compare the following outcome between two independent groups:

anesthesia time

time to surgical readiness

surgical time,

emergence time,

and total operating room time

I am wondering should I use survival analyze or independent t test? I know survival is for time to event data and censored data, and here as you see my outcomes variables is related to time. however I think it is possible to compare the mean of these outcome using t -test

can I say if I have a censored data I use survival analyze, otherwise I will use the simplest way which is t-test to do comparison?

Thanks.

Hi everyone. I want to DNA isolate from transfected cell culture medium (transfected with lentivirus). I tried lots of kit but DNA cons and quality not very well and I didnt show at agarose gel.

I am staining for VGLUT2 in mouse brain sections. I am seeing a lot of debris on the slices and these are visible in both Green and Red channels. By looking at the sizes I am sure that these are not my target signals. What can be the source of it.

I am currently working with HT-22 cells by transfecting them with bacterial plasmids to perform IHC. However, we have not had much success with reaching a 50% transfection rate. I usually do about a 40 hour incubation post transfection. I can not allow the cells to incubate any further because they will die due to the lipofectamine. Last week we tried a 60 hour incubation post transfection to have a better transfection rate. Unfortunately, we did have more transfected cells but many dead cells as well which defeats the purpose of our transfection. The one thing that did help us get a higher transfection rate was adding more plasmid, lipofectamine and P3000 to our cells. Does anyone have suggestions on how to improve the transfection rate without exceeding the 40 hour incubation?

P.S. I have attached files related to the products I use for my transfection and the protocol.

Hello Everyone !

I isolated mixed cells from neonatal mouse cortices , and they are always contaminated with other cells (by imaging it appears that my astrocytes are even the least common population) .. any ideas what I can do to prevent this ?

I plate on pdl coated flasks , and shake the mixed cells when confluent (1 hr on 180 rpm to remove microglia) and (4-6 hrs on 250 rpm to remove opcs)