May 31, 2015
SUBMITTED TO:
Dr.RIAZ HUSSAIN PASHA
SUBMITTED BY:
M.JUNAID SOHRANI11-arid-959M.UMAR11-arid-953WAQAS NAWAZ11-arid-975
Systemic Veterinary Histology
Topic
o WHAT IS STAINING???????????.....o STAINS USED IN LIGHT
MICROSCOPY……
Cellular Staining
Cell staining is a technique that can be used to better visualize cells and cell components under a microscope
By using different stains, one can preferentially stain certain cell components such as a nucleus or a cell wall, or the entire cell Most stains can be used on fixed, or non-living cells some can be used on living cells
some stains can be used on either living or non-living cells
Why Stain Cells?????
most basic reason that cells are stained is to enhance visualization of the cell or certain cellular components under a microscope
Cells may also be stained;
to highlight metabolic processes to differentiate between live and dead cells in a sample
Cells may also be enumerated by staining cells to determine biomass in an environment of interest
How Are Cells Stained and Slides Prepared?
Cell staining techniques and preparation depend on;
the type of stain analysis used
One or more of the following procedures may be required to prepare a sample
Permeabilization
Fixation
Mounting
Staining -
Acid Fuchsin
Aniline Blue
Basic Fuchsin -
Crystal Violet (Gentian Violet)
Eosin
Fast Green
Geimsa Stain
Hematoxylin
Methylene Blue Chloride
Safranin Toluidine Blue
Specific stains for Light Microscopy
Common Stains
There are several types of staining media
each can be used for a different purpose
Commonly used stains and how they work are listed below:
Bismarck Brown Carmine Coomassie blue Eosin DAPI Crystal violet Hematoxylin Ethidium bromide Fuchsin Hoechst stains Iodine Malachite green Neutral/Toluylene red Nile blue Methylene blue Nile red/Nile blue oxazone Osmium tetroxide Rhodamine Safranin
All these stains may be used on fixed;non-living cells living cells
Bismarck Brown - colors acid mucins, a type of protein, yellow and may be used to stain live cells
Carmine - colors glycogen, or animal starch, red
Coomassie blue - stains proteins a brilliant blue, and is often used in gel electrophoresis
Crystal violet - stains cell walls purple when combined with a mordant. This stain is used in Gram staining
DAPI - a fluorescent nuclear stain that is excited by ultraviolet light, showing blue fluorescence when bound to DNA. DAPI can be used in living of fixed cells
Eosin - a counterstain to haematoxylin, this stain colors red blood cells, cytoplasmic material, cell membranes, and extracellular structures pink or red.
Ethidium bromide - this stain colors unhealthy cells in the final stages of apoptosis, or deliberate cell death, fluorescent red-orange.
Fuchsin - this stain is used to stain collagen, smooth muscle, or mitochondria
Hematoxylin - a nuclear stain that, with a mordant, stains nuclei blue-violet or brown.
Hoechst stains - two types of fluorescent stains, 33258 and 33342, these are used to stain DNA in living cells.
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.
Neutral/Toluylene red - stains nuclei red and may be used on living cells
Nile blue - stains nuclei blue and may be used on living cells.
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
Safranin - a nuclear stain used as a counterstain or to color collagen yellow
After staining cells and preparing slides, they may be stored in the dark and possibly refrigerated to preserve the stained slide, and then observed with a microscope
Thank you