Staining

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