Iron Hematoxylin Methods
Verhöeff's method, which is one of the most popular techniques, requires that attention be focused on one type of fibre at the expense of the others as it cannot demonstrate the coarsest and finest fibres at the same time, requiring a choice as to which are to be optimally stained. Fortunately, in most practical applications, the elastic fibres in any single section are usually similar in size and the staining of each section may be optimised for the fibres it contains. Should it be necessary to demonstrate thick and thin fibres in the same section then a method other than Verhöeff's may be more satisfactory.
Verhöeff's method has become popular because it is fairly rapid, taking perhaps 60-90 minutes to perform. The elastic fibres are darkly stained (black) and stand out well in contrast to other tissues, which may be yellow-brown or red from a Van Gieson counterstain. It also photographs well. Its major disadvantage is that it is not an automatic method, that is, its success depends on the skill of the person performing the technique, although that is not difficult to do.
Musto's method is a modification of Verhöeff's, but stains from a low pH solution containing hydrochloric acid. This makes the solution progressive, and differentiation of the elastic fibres is not required. Musto recommended a modification of the HPS method, but replaced phloxine with woodstain scarlet. A variation using a Van Gieson counterstain was also given.
Musto's approach is not unusual, as several methods incorporating elastic fibre stains, often based on Verhöeff's hematoxylin counterstained with HPS or other trichrome variants, have been suggested under the general term of pentachrome or Movat methods, some with and some without mucin staining. Fundamentally, they are an elastic fibre stain with a trichrome counterstain of some kind, either of the Masson type or of the HPS type. The Silverman-Movat technique is similar but uses orcien to stain the elastic.
|Iron Hematoxylin Staining Methods|
|Musto – HPS variant|
|Musto – Van Gieson|
Last updated January 2019