Bacterial membrane staining
-
Last Update: 2021-01-21
-
Source: Internet
-
Author: User
Search more information of high quality chemicals, good prices and reliable suppliers, visit
www.echemi.com
(i) Experimental purpose
: learning the bacteria's membrane staining method:
(ii) experimental principle:
because of the affinity between the diaphragm and dye, not easy to color, usually using negative staining method to dye the membrane, that is, try to make the bacteria and background coloring and membrane does not color, so that the membrane around the bacteria body is a transparent circle. Because the water content of the film is more than 90%, so dyeing is generally not
heating
fixed, so as to avoid the film wrinkle deformation.
(iii) the test equipment
1. Living materials:
culture
glial bud bacillus (Bacillus mucilaginosus, commonly known as "potassium bacteria") for 3-5 days. When the bacteria grow on the carbon-based
of glycol
, the membrane is abundant.
2. Staining fluid and
reagents
:Tyler staining fluid: (see annex II, (i), 14), drawing ink after
filtration paper
filtration
and red staining fluid (see Annex II, (i), 2), melanin (see Annex II, (i), 7), 6% glucose aqueous solution, 1% methyl purple aqueous solution, methanol, 20% CuSO4 aqueous solution, balsamic asphalt, xylene.
3. Equipment: slides, slide shelves, wipe paper,
microscopes,
etc.
(iv) Experimental method
recommends the following four dyeing methods, of which the wet ink method is simpler and suitable for a variety of bacteria with pods. This is even more effective if examined with a difference microscope.
1. Negative staining method:
(1) production: take a piece of clean slide, add a drop of distilled water, take a small amount of bacteria into the water droplets mixed and coated.
(2)
dry
: dry the smears in the air or dry them with a dry air blow.
(3) staining: add red staining liquid staining 2-3min on the coating surface.
(4) wash: wash the red dye with water.
(5) dry: put the staining sheet in the air to dry or dry with a cold air blow.
(6) Melanin: Add a small drop of melanin to the left side of the dye coating surface, gently touch the melanin with an edge smooth slide, spread the melanin along the edge of the slide, and then drag to the right to make the melanin a thin layer on the dye coating surface and quickly dry.
(7) mirror inspection: first low mirror, then high mirror observation.
This article is an English version of an article which is originally in the Chinese language on echemi.com and is provided for information purposes only.
This website makes no representation or warranty of any kind, either expressed or implied, as to the accuracy, completeness ownership or reliability of
the article or any translations thereof. If you have any concerns or complaints relating to the article, please send an email, providing a detailed
description of the concern or complaint, to
service@echemi.com. A staff member will contact you within 5 working days. Once verified, infringing content
will be removed immediately.