DigsFish Services Recipes Page
Wet smears/blood smears/heart smears/digestive gland smears
Use the DiffQuick Stain Kit (Lab Aids, 3 Gondola Rd, Narrabeen Victoria 2101 phone (02) 9913 1396, fax (02) 9659 9559)
all fixatives contain toxic substances which can be harmful to humans. Please check the material safety data (MSDS) sheets which are available for each of the components of these fixatives and take appropriate precautions whenever they are being handled and used. MSDS data can be found online at MSDS.com
. For all fixatives, ensure that the volume of fixative used is at least 5 times that of the tissues being fixed.
Neutral buffered 10% formalin
(for histology - freshwater fishes)
100 ml formalin, 900 ml tap water, 4g sodium dihydrogen phosphate, 6g disodium hydrogen phosphate pH 7), optimum tissue thickness 4-5 mm.
10% Seawater formalin
(for histology, marine fishes and molluscs)
100 ml formalin, 900 ml of filtered seawater (filter to 10µm or less), optimum tissue thickness 4-5 mm.
(for histology, crustaceans)
110 ml of glacial acetic acid, 220 ml of formalin, 330 ml of 95% ethanol, 340 ml distilled water. Optimum tissue thickness 4-5 mm. Inject fixative into the hepatopancreas of crustaceans to ensure rapid fixation.
(for DNA analysis)
737 ml of 95% (or 700 ml of 100%) ethanol, 300 ml of distilled water. Optimum tissue thickness approx. 1 mm.
(for electron microscopy, store at 4°C)
4 ml of 50% gluteraldehyde, 96 ml of filtered seawater (filtered to 0.22µm) - optimum fixation time 24 hours at 4°C. Optimum tissue thickness approx. 1 mm
Use Simport M492 histology cassettes for large specimens, or Simport M493 cassettes for very small specimens.
Needles and syringe combinations for fish of various sizes (please dispose of needles thoughtfully):
5-20 cm fish - 25g x 5/8 inch needle, 1 ml syringe
20-60 cm fish - 21g x 1½ inch needle, 3 ml syringe
>60 cm fish - 18g x 1½ inch needle, 3 ml syringe
Follow manufacturers instructions to make up thioglycollate (29.3 grams of powder to 1 litre of freshwater). Add 20 g of salt (NaCl) to each litre of thioglycollate when using freshwater.
Stir then sterilise thioglycollate and universal tubes in an autoclave or pressure cooker (121°C for 15 minutes). Cool down to body temperature then add antibiotic/mycotic solution (50 ml for each 1 L of thioglycollate). Then dispense 10 ml aliquots into sterile universal tubes. Store the sterile tubes and media in the dark until use.
(for identification of Perkinsus
1 gram of iodine granules, 2 grams of potassium iodide
added to 100 ml of freshwater
DiffQuick Stain Kit
Lab Aids, 3 Gondola Rd, Narrabeen Victoria 2101 phone (02) 9913 1396, fax (02) 9659 9559
Fixative reagents and equipment, histology cassettes, needles and syringes:
Australia - all equipment , try Biolab (phone 1300 735 292, fax 1800 067 639) email Biolab.com.au
New Zealand -
- Global Science (phone 0800 734 100, fax 0800 999 002) email Global Science
- Biolab Scientific (0800 933 966) email Biolab.com
- Formalin (37% formaldehyde solution) Andrew Industrial Auckland (09) 444 0935
- Leak proof containers (e.g. cat #6235 from Stowers Containment Solutions (04) 939 9005
- 1.5 ml eppendorf tubes, microscope slides - Global Science
- Potassium iodide, Iodine granules - Global Science
- Antibiotics/antimycotics - (GibcoBRL 15240-062, 100 ml)- Global Science
- thioglycollate - (Oxoid CM173B, 500g) - Global Science
- Anaesthetics AQUI-S.com