23 Questions 36 Answers 0 Followers
Questions related from Mikael Niku
I'm looking for a green DNA counterstain for immunofluoresence. SYBR Green looks promising (except that apparently it fades quickly) but has anyone tested it?
30 October 2019 5,923 3 View
Which RNA extraction method would you recommend for RNA-Seq of snap-frozen fetal murine tissues? We will be doing Lexogen QuantSeq. Considering Qiagen RNeasy (good, but pretty expensive) or Thermo...
03 October 2019 9,704 3 View
Does anyone know a resource for data on organ sizes (mass, preferably also dimensions) for fetal mice at various stages of development?
15 March 2019 2,406 1 View
RNA extraction from laser microdissected FFPE tissues is markedly enhanced by sonication (paper below). However, I could only find a kit and protocol for Covaris sonicators. We only have a...
21 December 2018 3,115 1 View
We are trying to detect gene amplification in canine tissue sections (histomorphology is important). The amplified region is about 60 kb in length. What is the best way to prepare the probe,...
04 April 2017 8,921 2 View
Is there any way to detect/measure activation of TLRs (preferably specific TLRs) in tissue sections?
12 January 2017 4,616 3 View
We are happily extracting bacterial DNA using a bead beating protocol. Now we need to start processing swab samples containing fecal etc bacteria. If we put the entire swab into the bead beating,...
27 June 2016 555 4 View
I'm trying to analyse images of animal tissues produced with a 3DHistech slide scanner. I usually use ImageJ but have problems fitting large enough images in the memory. Are there any other free...
27 April 2015 3,475 18 View
I'm trying to visualize intestinal bacteria in paraffin sections. The problem is, the bacterial cells seem to shrink so much that no shapes are preserved, they are all just tiny roughly circular...
21 April 2015 7,139 12 View
Hello! What is the best histological stain or immunostaining marker to visualize bacteria in sections of animal tissues? Optimally, staining all (or most) bacteria but no eukaryotic cells, and...
27 October 2014 2,918 6 View
I would be especially interested in the color quality of the images. I'm trying to find a new camera for our histology teaching laboratory, and in this case we need live full HD image with good...
04 December 2013 9,082 2 View
I'm having trouble with contamination of linearized plasmid with the intact circular plasmid. I have heavily overdigested the plasmid, gel purified, and digested with another restriction enzyme,...
26 November 2013 2,061 5 View
I'm in the process of optimizing plasmid transfection to human K562 cells but not very happy with the success rate so far. Any tips or tricks here? Have you tried whether it is better to have the...
01 November 2013 3,670 2 View
I would like to measure immunofluorescence signal intensity per cell, using ImageJ or other readily available software. I have previously used particle detection to measure sizes etc., but this...
03 September 2013 6,635 15 View
I'm in the process of setting up IF staining for 5-methylcytosine (clone 33D3) and 5-hydroxymethylcytosine (the ActiveMotif polyclonal). Most protocols (and the manufacturer's instructions) use...
02 September 2013 3,104 3 View
I need to count a lot of cells in IF images where the stain is on the membrane (so the objects are circles rather than solid blobs like nuclei). This is tissue, so the cells are also clumped....
12 June 2013 6,795 19 View
Has anyone tried violet fluorescence dyes (such as DyLight 405, maximum emission at about 420 nm) in standard epifluorescence microscopy? It's in the visible range but I guess the sensitivity of...
26 April 2013 2,350 14 View
I'm using sodium butyrate in cell culture, but I'm slightly worried about the stability of my reagents. There is very conflicting information available regarding its stability in aqueous or...
11 March 2013 6,707 2 View
Hello! May I ask for your recommendations for tissue processing methods for laser microdissection and subsequent RNA isolation? I can think of at least the following protocols, each with...
13 February 2013 1,702 15 View
I wondering which of these labels is better for standard epifluorescence. I have been happily using Alexa546, but I recently got some fading problems, and someone recommended me to try Alexa555...
13 February 2013 9,860 9 View
I have problems getting very weak fluorescence signal from my probe-based RT-qPCR assay. The Ct is not bad (less than 30, and I see a clear although weak band on gel), but the signal intensity is...
18 January 2013 1,934 3 View
I have run into a strange problem: my immunofluorescence stainings, done on PFA-fixed paraffin sections using an Alexa546 labelled secondary antibody and mounted using Dako Immunofluorescence...
16 January 2013 4,567 14 View
We are planning to do RNA-Seq for RNA extracted from two types of samples: Routine snap-frozen mouse fetal tissues Laser microdissected tissue sections (FFPE sections and/or cryosections) We only...
01 January 1970 906 2 View
