Annals of the Royal College of Surgeons of England
(I977) vol 59
INSTRUMENTS AND TECHNIQUES
Differential staining of thin brain slices D H Tompsett PhD Past Prosector to the Royal College of Surgeons of England
The method of staining the grey matter of human brain slices after inhibiting the staining of the white matter by soaking each slice in Mulligan's hot phenol solution is well known and comparatively easy. I include this technique with modifications, especially of the method of mounting and storing the slices, in the second edition of Anatomical
Techniques'. I found that the most convenient thickness for each slice, to minimize the risk of some slices fragmenting, was i cm. An average adult brain provided between IO and I 2 slices, which was a convenient number to mount as wet specimens in a box rather like a giant slide box. When not in use the slices are kept in darkness as the blue surface stain gradually
the thick ones. In spite of this defect the thin slices were adequate for study so they were mounted on a single sheet of Perspex and stored in a light-proof box. A year before retirement recently I noticed that the thin slices had faded completely so I decided to prepare two new sets of thin brain-stem slices. I stained the first test slice exactly as I had always stained the thick slices; as I expected, the staining was very poor. As the hot phenol solution is used to inhibit overall staining the most logical experiment seemed to be greatly to reduce the time (5 min for i-cm slices) in which each thin slice was immersed in the phenol. I found that the ideal time for treatment in the phenol soilution was about 5 s. Solme slight adjustment may be needed in the time in which thin slices are soaked in the other solutions, but this makes so little difference to the staining that it is of little consequence. One set of thin brain-stem slices has now been mounted. It bears favourable comparison with any of the thick slices.
fades with prolonged exposure to bright light. However, slices prepared by this method have faded only very slightly after 25 years. When I made the first museum series of slices i cm thick I also made two similar series of slices of the brain stem only, which varied in thickness (through human error) between 2 and 3 mm. I stained the thin slices by exactly the same procedure as used for the thick ones. I was disappointed to Reference see that the staining of the thin slices was I Tompsett, D H (1970) Anatomical Techniques, 2nd edn. Edinburgh and London, Livingstone. very inferior to the corresponding areas of
(I977) vol 59
INSTRUMENTS AND TECHNIQUES
Differential staining of thin brain slices D H Tompsett PhD Past Prosector to the Royal College of Surgeons of England
The method of staining the grey matter of human brain slices after inhibiting the staining of the white matter by soaking each slice in Mulligan's hot phenol solution is well known and comparatively easy. I include this technique with modifications, especially of the method of mounting and storing the slices, in the second edition of Anatomical
Techniques'. I found that the most convenient thickness for each slice, to minimize the risk of some slices fragmenting, was i cm. An average adult brain provided between IO and I 2 slices, which was a convenient number to mount as wet specimens in a box rather like a giant slide box. When not in use the slices are kept in darkness as the blue surface stain gradually
the thick ones. In spite of this defect the thin slices were adequate for study so they were mounted on a single sheet of Perspex and stored in a light-proof box. A year before retirement recently I noticed that the thin slices had faded completely so I decided to prepare two new sets of thin brain-stem slices. I stained the first test slice exactly as I had always stained the thick slices; as I expected, the staining was very poor. As the hot phenol solution is used to inhibit overall staining the most logical experiment seemed to be greatly to reduce the time (5 min for i-cm slices) in which each thin slice was immersed in the phenol. I found that the ideal time for treatment in the phenol soilution was about 5 s. Solme slight adjustment may be needed in the time in which thin slices are soaked in the other solutions, but this makes so little difference to the staining that it is of little consequence. One set of thin brain-stem slices has now been mounted. It bears favourable comparison with any of the thick slices.
fades with prolonged exposure to bright light. However, slices prepared by this method have faded only very slightly after 25 years. When I made the first museum series of slices i cm thick I also made two similar series of slices of the brain stem only, which varied in thickness (through human error) between 2 and 3 mm. I stained the thin slices by exactly the same procedure as used for the thick ones. I was disappointed to Reference see that the staining of the thin slices was I Tompsett, D H (1970) Anatomical Techniques, 2nd edn. Edinburgh and London, Livingstone. very inferior to the corresponding areas of
