Stains in MeTHOD pREATICAL Flashcards
What is Vivo Staining ?
the process of dyeing living tissues— in vivo means “in life” (compare with in vitro
staining).
How does it work ?
It works by letting certain cells or structures take on contrasting color(s), their form (morphology) or position within a cell or tissue can be readily seen and studied
It involves colouring cells or structures that are no longer living. .
What is the actual purpose ?
reveal cytological details that might otherwise not be apparent;
however, staining can also reveal where certain chemicals or specific chemical reactions are taking place within cells or tissues.
How are cells/Slides prepared ?
Permeabilization - treatment of cells, generally with a mild surfactant, which dissolves cell membranes in order to allow larger dye molecules to enter inside the cell.
• Fixation - serves to “fix” or preserve cell or tissue morphology through the preparation process.
but most fixation procedures involve adding a chemical fixative that creates chemical bonds between proteins to increase their rigidity.
What are common stains for fixation ?
Common fixatives include formaldehyde, ethanol, methanol, and/or picric acid.
What are some procedure in preparing sample ?
Mounting - involves attaching samples to a glass microscope slide for observation and analysis.
Staining - application of stain to a sample to color cells, tissues, components, or metabolic processes.
What is Bishmark Brown ?
Staining - application of stain to a sample to color cells, tissues, components, or metabolic processes.
List some stains and their function ?
Iodine - used as a starch indicator. When in solution, starch and iodine turn a dark blue color.
- Malachite green - a blue-green counterstain to safranin in Gimenez staining for bacteria. This stain can also be used to stain spores.
- Methylene blue - stains animal cells to make nuclei more visible.
Name one stain and its function ?
Ethidium bromide - this stain colors unhealthy cells in the final stages of apoptosis, or deliberate cell death, fluorescent red-orange.
- stains nuclei red and may be used on living cells.
Neutral/Toluylene red
Nile red/Nile blue oxazone
this stain is made by boiling Nile blue with sulfuric acid, which creates a mix of Nile red and Nile blue. The red accumulates in intracellular lipid globules, staining them red. This stain may be used on living cells.
Osmium tetroxide
- used in optical microscopy to stain lipids black.
Rhodamine -
a protein-specific fluorescent stain used in fluorescence microscopy. Muntjac skin fibroblast labeled with rhodamine phalloidin
Safrani
a nuclear stain used as a counterstain or to color collagen yellow.
Indirect Fluorescent Staining Methods
• This approach requires only the primary antibody
This approach requires only the primary antibody and a fluorphore-conjuated secondary antibody, and is the simplest form of signal amplification.