A Method for staining both Gram-Positive and Gram-Negative Baeteria in Sections.
WS Ollett - 1947 - cabidigitallibrary.org
WS Ollett
1947•cabidigitallibrary.orgThe author has found the existing methods for staining both Gram-positive and Gram-
negative bacteria in sections unsatisfactory: but has obtained excellent contrast by the use of
a modification of the TWORT method (J. State Med., 1924, v. 32, 351) in" which the light
green of Twort's stain is replaced by last green FCF to avoid fading. Material is fixed in 5 per
cent formol-saline, embedded in paraffin, sectioned at 3µ and brought down to distilled
water in the usual manner. The section is then stained for 3 to 5 minutes in aniline-crystal …
negative bacteria in sections unsatisfactory: but has obtained excellent contrast by the use of
a modification of the TWORT method (J. State Med., 1924, v. 32, 351) in" which the light
green of Twort's stain is replaced by last green FCF to avoid fading. Material is fixed in 5 per
cent formol-saline, embedded in paraffin, sectioned at 3µ and brought down to distilled
water in the usual manner. The section is then stained for 3 to 5 minutes in aniline-crystal …
Abstract
The author has found the existing methods for staining both Gram-positive and Gram-negative bacteria in sections unsatisfactory: but has obtained excellent contrast by the use of a modification of the TWORT method (J. State Med., 1924, v. 32, 351) in "which the light green of Twort's stain is replaced by last green F.C.F. to avoid fading.
Material is fixed in 5 per cent formol-saline, embedded in paraffin, sectioned at 3µ and brought down to distilled water in the usual manner.
The section is then stained for 3 to 5 minutes in aniline-crystal violet; this is poured off and the preparation is washed quickly in distilled water.
The section is treated with Gram's iodine for 3 minutes: this is poured off, the preparation is "washed quickly in distilled water and blotted dry. Decolourization is effected with 2 per cent, acetic acid in absolute alcohol: the section should now be a dirty straw-colour. A further wash in distilled water is made.
Counterstaining is made by Twort's neutral-red- fast-green, diluted 1 part with 3 parts of distilled "water, for 5 minutes. A final wash in distilled water is made. The section is then decolourized with 2 per cent, acetic-acid-alcohol until no more red stain comes away (15-30 seconds).
The preparation is cleared in xylol and mounted in D.P.X. or neutral balsam. Microscopically, nuclei are red, cytoplasm and red blood corpuscles light green, Gram-positive bacteria dark blue and Gram-negative bacteria pink. H. J. O'D. Burke-Gaffney.
CABI Digital Library