In the following powerpoint tutorial I run through how you can use ImageJ/FIJI to import your raw microscopy image files and then analyse and track individual cells to generate single cell fate maps. Note, unless you are also planning on quantifying fluorescent intensity data in your movies, I strongly suggest converting all of your image files to high quality JPEGs…
Here is a step-by-step guide that I made to help people export microscope images from ImageJ/FIJI and then import and alter colours/levels etc in Photoshop. The guide also shows you how to easily move from Photoshop to illustrator to make montage images etc for publication. PDF Download: Guide to FIJI-Photoshop Image manipulation
I often get asked how to uses Thresholds to measure things in Image J. There are some great guides on the web explaining how to use Thresholds in Image J, and here are a few that are well worth checking out [Link1][Link2]. Below are some of the Basic Steps for using Thresholds: Open your image and duplicate it (Image>Duplicate) On the…
Image J can be downloaded for free from here . This guide can also be downloaded as a complete PDF here: Measuring Cell Fluorescence using ImageJ Here is a very simple guide for determining the level of fluorescence in a given region (e.g nucleus) Select the cell of interest using any of the drawing/selection tools (i.e. rectangle, circle, polygon or freeform) From the Analyze menu…
The Mitchison Lab has an excellent guide on staining and fixing cells for Actin and Microtubules which is worth reading [Link] Coverslips Most coverslips come with a fine film coating to stop them sticking to each other. This can reduce the ability of coating agents such as poly-L lysine from working properly, and can thus reduce the ability of cells to…
General Guidelines These guides are written primarily for HeLa cells, but it should be possible to extend these to other cell lines with a bit of optimisation. 1) Make sure your cells are happy ! 2) Aim for around 75-85% confluence at the time of release. 3) Try and keep everything close to 37ºC, including media, washing media, PBS, TC…
Grow cells in DMEM (Sigma #D9443) without L-Arg, L-Leu, L-Lys ! This DMEM is low glucose 1000mg/L so you will need to add and extra 3000mg/L to make it unto 4000mg/L in total. Thus 15ml of the 10% Glucose solution is needed for 500ml of DMEM. Need to add 50ml of Mass Spec Grade Serum for final [conc.] of 10…
A complete ‘ish’ guide to Flow cytometry or FACS for cell cycle work. Covers all aspects from tissue culture, through to running and analyzing your samples on a FACScan or FACS Calibur. It was written by me a few years ago and so is a bit out of date. For example there is a new “prettier” version of CellQuest (Pro)…
Taken from Addgene's guidelines [Link] Antibiotic Concentrations The following list shows recommended antibiotic concentrations for LB media or agar plates. Antibiotic Concentration Ampicillin 100 μg/mL Bleocin 5 μg/mL Carbenicillin 100 μg/mL Chloramphenicol 25 μg/mL Coumermycin 25 μg/mL Gentamycin 10 μg/mL Kanamycin 50 μg/mL Spectinomycin 50 μg/mL Tetracycline 10 μg/mL Example: To make 100 mL of LB/ampicillin growth media, add 100 μL…
With the later versions of Photoshop CS4 and CS5 extended it is now very easy to add a scale bar to your microscope images. But before we go ahead you will need some information first. 1) The actual Pixel size of the camera attached to your microscope (here is a short list of some common cameras). 2) Did you use any…