00:01
In the direct immunofluorescence test, there is a biopsy
section or other tissue section placed on a slide.
00:15
A fluorescein-labeled sheep antibody
against the antigen of interest is added.
00:22
So for example, one might want to see whether a particular
patient in a biopsy was expressing a particular tumor antigen.
00:31
And one could use a sheep antibody
specific for that tumor antigen.
00:37
Under ultraviolet light, the fluorescein label
emits a visible light over the antigen of interest.
00:45
So for example, if we are looking for
a specific tumor antigen, then if
the patient expresses that tumor
antigen, there will be light emission.
00:54
If they do not, there
will be no light emission.
00:58
In the indirect immunofluorescence test,
the tissue can be of animal or human
origin as long as it contains the
antigen that one is interested in.
01:10
Patient serum is added, and in this
particular example where we are
looking for autoantibodies, the
autoantibodies bind to the antigen.
01:21
So this could be a tissue section of
thyroid for example, and we’re asking
the question: does the patient have
autoantibodies to thyroid antigens?
A fluorescein-labeled sheep anti-human IgG is added
and this binds to the patient’s immunoglobulin gene.
01:43
The bound IgG is detected
under ultraviolet light.
01:48
One example of this would also be
the measurement of anti-nuclear
antibodies in patients with systemic lupus erythematosus.
Here we have patient’s serum that
is incubated with a section of
rat liver, a section of rat kidney and a section of rat stomach.
02:13
Binding to the antigens present in these tissues is detected
by a fluorescein labeled anti-human immunoglobulin antibody.
02:22
And we can see the patient is indeed positive
for these anti-nuclear antibodies where we
can see fluorescence of a liver section, of
a kidney section and of a stomach section.
02:35
As well as these immunofluorescence techniques,
one can also use immunohistochemistry,
where a tissue section on a glass slide is
incubated with an enzyme-labeled antibody.
02:52
Binding will occur if the antibody
detects antigens in the tissue section.
03:00
The substrate for the
enzyme can be added.
03:05
The enzyme mediates a color
change in this substrate.
03:10
And this can be detected
under the microscope.
03:18
In indirect immunochemistry, the patient’s antibody is
detected using an enzyme labeled anti-human immunoglobulin.
03:28
Again the enzyme mediates a color change in the
substrate which can be visualized under a microscope.