J.Jpn. Surg. Soc.. 57(9): 1542-1547, 1956

---

HISTOCHEMICAL STUDIES OF GALL STONES: PARTICULARLY IN RESPECT OF THE PROTEIN CONTENT

Histochemical studies were carried out as a method of analytical study of gall stones. In this paper a specific technique for the preparation of serial sections of gall stones and their complete decolorization were discussed. Employing this new technique, protein which has hitherto never been detected, was demonstrated in the gall stone.
(A) Method
(I) Preparation of serial sections of the gall stone :
(a) A gall stone which was cut in a half was embedded in paraffin.
(b) Because of the fragility of the stone, the cut surface was made smooth and a thin sheet of paraffin paper was adhered to the cut surface with balsam or with acacia.
(c) Serial sections with 15 to 40 microns were made by a microtome. Procedures (a) and (b) were repeated for each section.
(d) A section was placed on a cover glass with the cut surface facing to the glass, and was fixed to the glass with plastic or by egg white, and then dried.
(e) The paper was removed by immersing it in xylol or in water.
(II) Decolorization:
(a) A cover glass with the section was completely dried and left in xylol for 24 hours, then dried.
(b) This was then left in a mixture of absolute alcohol and carbon tetrachloride in equal volume at room temperature or at 60 °C for 1 to 2 days. If the decolorization was not sufficient the same procedures were repeated. Washed with absolute alcohol, and dried.
(III) Identification of protein:
(a) Decolorized specimen was left in absolute alcohol for over 5 minutes.
(b) The glass with the section was left in 0.1% absolute alcohol solution of potassium salt of tetrabromphenolphthaleinethylester (TBP) for 10 to 20 minutes.
(c) Washed with alcohol and then with N/5 acetic acid.
(d) TBP bound with protein shows bluish discoloration, whereas remaining substance are stained yellow.
(B) Results
(1) Gall stones rich in pigments do not necessary contain high per centage of protein.
(2) Protein was most rich in the peripheral or outer layers in pigment stones, whereas it was rich in the central layers in cholesterol stones.
(3) Almost no stone was found in which protein was most highly contained in midlayer in any type of stones.
(4) Generally, protein content in the central portion was much less when it was compared with the amount of fibrin.
(5) Protein contained in gall stones exhibit a marked alginin reaction, which is suggestive of the existence of histone-like protein.
These findings may suggest that gall stones are not formed in laminated manner a round a nucleus as it has been postulated by Aschoff, Bacmeister, Schade and others, but formed by organization of liquid form materials or coacervates which derived from disintegrated bile components due to inflammatory process and/or metabolic abnormalities.
Thus it may be considered that the gall stone is formed in one stage by a colloid chemical mechanism instead of the theory postulated by Aschoff and others.
(author's abstract)

---

---

To read the PDF file you will need Adobe Reader installed on your computer.