How to analyse single cells time-lapse movies using Fiji and/or Photoshop

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 […]

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User guide: Export microscope images from ImageJ/FIJI into Adobe Photoshop and Illustrator

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

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Immunofluorescence Guide

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 […]

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HeLa SILAC conditions

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 […]

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Antibiotic Concentrations for Bacterial Work

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 […]

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