29 Questions 6 Answers 0 Followers
Questions related from Sebastian Freeman
Greetings everyone, Would anyone be able to suggest or know of software to model the dosage of UV received by a surface from a source object? For example, I would like to model how a cylindrical...
16 May 2020 3,402 2 View
Greetings everyone, My lab, as with others many others, have shifted some of our efforts to aid in the global fight against COVID-19. As part of our efforts, we are looking to evaluate current...
07 May 2020 612 0 View
Hello everyone, I was wondering whether anyone had experience getting thin slices of PDMS. The PDMS samples we want to examine start off already being 0.8 - 1.5 mm in thickness, but we would like...
29 August 2018 2,702 6 View
Hello everyone, I am trying to troubleshoot my SEM procedure. I am trying to image gelatin gel microstructure using SEM. These gels extruded out of a small diameter nozzle (0.33 mm ID). I have...
31 July 2018 6,049 9 View
Hello everyone, I would like to know what buffer solution is best for maintaining bovine thrombin activity if I intended to store the thrombin solution frozen at -70 C. I do intend to use the...
13 June 2018 809 3 View
Hello everyone, I would like to infect my NIH-3T3 cells with CMV promoter lentiviral particles to induce GFP expression. The particular plasmid is from Addgene (https://www.addgene.org/17446/)....
17 July 2017 8,538 0 View
Hi everyone, When culturing a hydrogel in a well-plate, how do you decide how much medium to put over the gel? I have noticed that for 2D culture, the recommendations for medium volume give...
04 May 2017 3,118 3 View
Hi everyone, I am working with 3T3 fibroblasts currently and mixing them together into a hydrogel composed of fibrin, gelatin, and glycerol. I performed a live/dead cytotoxicity assay on them, and...
10 April 2017 9,955 3 View
Hello everyone, In terms of methods, should one increase concentration of antibody solution or increase the volume of incubation when the signal is weak? This could apply to the primary or...
01 February 2017 7,558 3 View
Hi everyone, Could anyone who has experience with performing whole gel immunostaining of 3D cell cultures share some tips and troubles they have had when optimizing their protocols. I work with...
01 February 2017 2,862 1 View
Hello everyone, I was hoping someone might have some insight into a problem I have with filtering fibrinogen solutions. I use bovine fibrinogen from Sigma Aldrich (F8630), which is Type I-S,...
15 December 2016 2,354 5 View
Hello everyone, I am trying to find out if any research has already been done on the study of pro-angiogenic factor expression by cancer cells in 3D cell culture. In interested specifically in...
01 December 2016 4,010 2 View
Hello everyone, I am interested in immunostaining cell-laden hydrogel scaffolds for secreted proteins, such as VEGF. I can do both whole scaffold immunostaining and also cryosection. I typically...
06 October 2016 6,313 0 View
Hello everyone, In my lab we have an epifluorescence microscope. Since we purchased the filter cubes from Chroma, I often go to the Chroma Spectra Viewer (https://www.chroma.com/spectra-viewer) to...
03 October 2016 9,344 1 View
Hello everyone, I often grow endothelial cells in 3D gels, but one thing I would like to be certain of is whether or not they are developing a lumen. I have read several papers which show images...
20 September 2016 7,250 5 View
Hello everyone, I often work with hydrogels and in many cases I have small pieces of plastic tubing embedded in the hydrogel. I am not very well versed in the terminology around this subject,...
16 September 2016 8,198 3 View
Hi everyone, I have a question about accepted definitions of terminologies when it comes to hydrogels such as collagen, gelatin, alginate, etc. I have heard and read the two terms gelation and...
03 August 2016 226 3 View
Hi everyone, I have recently become interested in using the transglutaminase enzyme for the crosslinking of protein-based scaffolds. I have seen the name "Moo glue" cited in a few papers, and I...
05 May 2016 1,252 0 View
Hello everyone, I am looking in particular for MCF-7 breast cancer cells and HUVECs already transfected to express fluorescent proteins such as GFP. I would prefer the HUVECs at as a low a passage...
02 March 2016 3,906 3 View
Hello everyone, I wanted to form a solution of a mixture of rat tail collagen type I and 600 Da PEGDA. I kept everything on ice except the PEGDA, and added a little bit of phenol red into the...
26 February 2016 7,578 0 View
Hello everyone, I was particularly inspired by the paper by Antoine, Vlachos, & Rylander 2015 - Tunable collagen I hydrogels for engineered physiological tissue micro-environments. In it, they...
15 December 2015 8,167 8 View
Hello everyone, I was culturing a cancer cell line and primary HUVECs for an experiment for immunofluorescence. I had treated the plastic surface with poly-L-lysine for improved adhesion since in...
27 November 2015 7,726 6 View
Hello everyone, I have been preparing cryosectioned tissue samples of mouse and rat spleens and tumors. I followed the advice of some researchers from here to fix as soon as the tissue was...
19 November 2015 6,738 4 View
I would like to know if there is a simple approach to show whether or not a primary polyclonal antibody is not specific enough for an immunofluorescence experiment? I am using a anti-PECAM1...
14 November 2015 4,715 4 View
Hi everyone, I currently use a slightly modified version of Rajan et al.2006 for preparing type I collagen solutions from rat tail tendons in lab. I was wondering if anyone knew any other...
29 October 2015 9,372 0 View
Hello everyone, I am trying to label two different antigens in two different cells of interest, and I want to also be able to label them when the cells are cultured together. I am trying to figure...
16 September 2015 9,266 3 View
Hello everyone, My lab regularly extracts collagen from rat tails of normal rats, and we would like to include steps for sterilization. I have seen some protocols but I am unsure what step or...
21 August 2015 3,165 3 View
Hi everyone, I'm interested in flow through a collagen scaffold. I am thinking of using needles to insert into the collagen scaffold hooked up to a peristaltic pump. Has anyone done this before...
18 August 2015 2,965 3 View
Hi everyone, I am designing a cell culture vessel for my experiments and I have come upon a design consideration that I am not sure how to proceed with, and that is, what would be an adequate...
01 January 1970 5,587 2 View
