what is DAPI colour and what is its antitbody
blue and binds to nucleai
if nucleai are centrally located what does it mean
it means the fibres are being regerated
which anitbody fluoreses and which one binds to the antigen
1 antibody attaches to the antigen and 2 antibody attaches onto 1 antibody and causes it to fluroes
would we see centrally located nucleai during exercise
no
what is the advantage of immunofluorescence
you can add many different 1 antibody immunofluorescene to the same sample
if we see increased collagen red what does this mean about muscle health
fibrosis, this is where there is collagen build up between primary cell types, this isn’t good
using DAPI can you tell the difference betwen nucleai and satelite cells
no
what is brightfield/ colorimetric staining
this is where sections are stained with a dye specfifc to certain proteins/ cell strcutures
what is MHY2 colour and what does is its antitbody
red and binds to type 2a muscle fire
what is Laminin colour and what does is its antitbody
green, binds to basal lamina and cell membrane
what does hematoxylin-eosin stain for
hematoxylin = nucleai
eosin = cytoplasm
if we use immunofluorescence what do we need to see it and can we see it under brightfield setting on a microscope
you need fluorescent light and you can not see it under brightfield setting
how do we stain for satelite cells
we use CD56 and Pax7 and look for overlay
how does electrophoresis work
samples are loaded into cells and shocked, bigger proteins move slower so it will end up at the top, smaller proteins move faster and end up at the bottom
does muscle biopsy effect a person ability to workout immediately after
no
are the skin and fascia frozen before a muscle biopsy
yes
why is the vastus lateralis typically chosen for muscle biopsy
it close to the skin and easy to access, no major arteries running though it
what does western blot measure for
it just allows you to quantify total protein in a sample
what are the 2 methodologies used to asses protein content
- histology
- western blot
what are the 2 types of staining procedures
- brightfield/ colorimetric
- immunofluroescence
what does picrosirius red dye for
sirus red = collagen red
picric acid = non-collagenous features
what is used to get quantification of muscle protein
electrophoresis
what setting do you need to use on a microscope in order to view brightfield/ colorimetric stains
brightfield setting
what are the disadvantages of histology
- muscle needs to be frozen right away
- you are looking at a small portion of the muscle
- you are making assumptions that the vastus lateralis is representetive of the whole muscle
