Alexander Chamessian

8 Questions 14 Answers 0 Followers

Questions related from Alexander Chamessian

What is the best way to store slide-mounted cryosections for ISH/IHC?

I cut thin sections (14 um) of fixed frozen brain and spinal cord tissues and mount on Superfrost Plus slides. While I'm sliding them, I let them dry for 10-15 mins at RT and then I store in -80C...

15 May 2018 550 6 View

How to best ship mounted cryosections?

I need to send a collaborator mounted cryosections of spinal cord tissue. The slides are stored at -80C here in the lab. What is the best way to ship? I've sent on dry ice, and that seems to work...

15 May 2018 7,863 3 View

Dissociating single cells from fixed tissue?

This is a long-shot question. I have some methanol-fixed tissue neural tissue. I'm wondering if it would be possible to get intact single cells from the tissue once rehydrated. I was thinking...

06 October 2017 3,862 4 View

How do you identify 'healthy' neurons in slices (brain, spinal cord) for Patch-Clamp recording?

I'm new to electrophysiology. I'm trying to record from dorsal horn neurons in the spinal cord. One of the hard things for me is identifying the neurons that are healthy and suitable for patching....

27 August 2017 4,737 9 View

Intraganglionic virus injection into the mouse DRG?

I'm needing to inject viral vectors directly into mouse DRG so that I can localize the expression just to the DRG. I can get very good expression when I inject intrathecally, but that enables the...

11 July 2016 2,511 0 View

Does Calmodulin bind M13 in the absence of calcium?

I am interested in the calmodulin interactions with its target peptides. One synthetic target peptide is M13, which is the basis for many GECIs such as GCaMP. My question is, in the...

03 May 2016 8,331 2 View

Plasmid-transformed Stbl3 grow well in 5 ml overnight culture (w/ antibiotics) but do not grow in 50 ml midiprep culture?

I am trying to do a routine midiprep of a plasmid for AAV production. The plasmid is pAd Delta F6 (AAV helper virus from UPENN). I got the plasmid directly from the core facility, transformed into...

19 February 2016 3,006 11 View

Has anyone done Cre-dependent shRNA in AAV?

I want to selectively knockdown some genes in specific neuronal populations using AAV in a Cre-dependent manner. FLEX or DIO vector are used often for transgene expression in this manner, but I...

23 October 2015 6,740 1 View