Editing Staining (section)

=== Types ===
{| class="wikitable"
|+ Types of staining techniques<ref>{{Cite book|title=Elementary Microbiology Vol - I}}</ref>
!Sr. No.
!Staining Technique
!Preparation
!Application
!Result
|-
|1.
|Simple (Monochrome)
|Smear stain with single dye .
e.g. Methylene blue, Safranin°≤×←→ etc.
|Used to highlight microbes and illustrate cellular
shapes and arrangements
|Organisms are stained in the color of applied stain
|-
|2.
|Negative (Relief)
|Smear mixed with Nigrosin and spread
into thin film
|Study cell morphology
|Organism is stained, the background is black
|-
|3
|Gram
|Primary stain: Crystal violet applied to film then treated with iodine (mordant), alcohol (decolourizer) and counter stained with safranin
|Characterizes bacteria in one of two groups, Gram positive or Gram negative
|Gram positive appears purple in color
Gram negative appears pink in color
|-
|4
|Acid fast (Ziehl-Neelsen technique)
|Film stained with hot Z.N.C.F. decolourised (acid-alcohol) and counter stain with methylene blue
|Separate non-decolorized acid fast bacteria that are not decolorized from colorized non-acid fast bacteria
|Acid fast bacteria:Red
Non acid fast: Blue
|-
|5
|Endospore (Dornor's method)
|Primary stain Malachite green heat fixed to penetrate spores; vegetative cells are counterstained with Safranin
|Detects the presence of endospores in six genera of bacteria
|Endospores: Green
Vegetative cells: Red
|-
|6
|Capsule
A: Hiss method (Positive technique)

B: Manevals's technique (Negative)
|Smear stained with Hiss stain following treatment with copper sulphate

Bacterial suspension smeared along with Congo red and the Maneval's stain is applied
|Capsules can be observed as clear zones surrounding cells of capsulated bacteria and are used to demonstrate the presence of capsules.
|Capsule: Light violet/pale mauve color
Bacteria: Purple capsule, bacterial cell, stands out against dark background
|-
|7
|Cell wall (Dyar's method)
|Smear treated with C.P.C. which dissociates to form positively charged cetyl pyridinium and negatively charged chloride ions. Positively charged ions are adsorbed on negatively charged cell wall
|Stains cell wall of bacterium
|Cell wall: Red Cytoplasm: Blue
|-
|8
|Flagella (Leifson's method)
|Mordant acts to thicken flagella before staining and increases visibility microscopically when stained with Leifson stain
|Demonstrates presence of flagella
|Flagella: Red Vegetative cells: Blue
|-
|9
|Nuclear material (Feulgen technique)
|Smear is treated for hydrolysis to release purines from DNA, purines to cause shift form furanose to aldehyde. Aldehyde groups are available to react with schiff's reagent to form addition compounds.
|To demonstrate the presence of DNA in cell. But for detection of the DNA, RNA should be selectively destroyed by acid hydrolysis without affecting DNA
|Nuclear material- pinkish purple,
Cytoplasm- colorless
|-
|10
|Metachromatic granules (Alberts's method)
|The smear is first treated with chloroform to remove fats . Smear applied with Alberts stain which contains cationic dyes such as toluidine blue and malachite green. Toluidine blue preferentially stains granules while malachite green stains cytoplasm.
|The granules show the typical monochromatism nature, this is used to demonstrate granules
|Granules: Bluish black, Cytoplasm: Green
|-
|11
|Intracellular lipids (Burdon's method)
|Lipids are stained with fat soluble dyes like Sudan black. On application of Sudan black-B dyes move into lipids and are retained there while cytoplasm is counter stained with safranin.
|To detect the presence of lipids in cell wall, cell membrane or fat globules (PHB in cytoplasm)
|Lipid granules: Deep blue,
Cytoplasm: Light pink
|-
|12
|Polysaccharide (Hotch kuss method)
|Polysaccharide is oxidized with periodate to form polyaldehyde which reacts with Schiff's reagents to red color, while cytoplasm is counter stained with malachite green
|Detects the accumulation of polysaccharide granules in the cells
|Polysaccharide: Red
Cytoplasm: Green
|}