Materials required

In order to carry out the protocols described on this site, you will need the following materials and equipment:

MaterialsDissecting tools

  • Dissecting kit: Fine forceps x 2, dissecting scissors, hair loop*

  • 3-4 petri dishes

  • Automated pipettes (e.g. Gilson, P20 and P200)

  • Correctly sized pipette tips

  • Microscope slides

  • Glass coverslips

  • Phosphate buffered saline solution**

  • Hoechst 33342 fluorescent DNA stain (see Protocols for details on preparation). Can be purchased from most scientific suppliers.

  • Eppendorf tubes / screw-tip microcentrifuge tubes

Equipment

  • Stereomicroscope (dissecting microscope) with gooseneck lighting. We use Leica S6 D and MZ7 stereomicroscopes.

  • Compound light microscope with the following: a fluorescence light source, BP 340-380 excitation filter and LP 425 suppression filter, and darkfield filter. We use a Leica DMLB FL and a Leitz Laborlux S.

  • Refrigerator for storing Hoechst and samples prior to examination.

 

*The hair loop is a homemade device, consisting simply of a strand of human hair, fashioned into a small loop and taped to either a cocktail stick or pipette tip to allow easy manipulation. It is simple yet incredibly effective!

**Phosphate saline solution can either be purchased from a scientific supplier in ready made sachets or tablets to be added to distilled water, or it can be made from scratch using the following recipe. The latter is much cheaper if you intend to use a lot of solution over a long period of time, but the former is more convenient for small projects and remote working.

Phosphate buffered saline from scratch

For 2 litres solution:

  • 2.3g di-sodium hydrogen orthophosphate
  • 16g sodium chloride
  • 0.4g potassium chloride
  • 0.4g potassium di-hydrogen orthophosphate
  • 2 litres distilled water

Salts are available to purchase from most scientific suppliers. Weigh out all salts (all come in powder form) and add to distilled water. Mix well until all salts have dissolved. Autoclave if sterility is required. Store in refrigerator at 1-5ºC.