Decal Histopath

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Nitric acid

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Most common and fastest

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Nitric acid

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Recommended conc. of 5-10%

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Nitric acid

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Inhibits nuclear stains and destroys tissues

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Prevent damage by nitric acid

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Combine w/ formaldehyde or alcohol

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Nitric acid

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Easily removed by 70% alcohol

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Nitric acid

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Imparts yellow color

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Formol-Nitric acid

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Less tissue destruction than 10% aqueous nitric acid

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To prevent yellow color imparted by nitric acid

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Add 5% NaSO4 and wash with running tap water for at least 12 hrs

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Make discoloration disappear

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Add 0.1% urea

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Perenyi's fluid

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Decalcifying and tissue softener

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Perenyi's fluid

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Maceration is avoided due to presence of chromic acid and alcohol

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Perenyi's fluid

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Complete decalcification cant be determined by chem test

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Prevent appearance of ppt in perenyi's fluid

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Add 0.5ml of saturated aqueous ammonium oxalate. Reappearance of ppt w/in 30 mins (decal is incomp)

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Phloroglucin-Nitric acid

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The most rapid among decal acids

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Phloroglucin-Nitric acid

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To remove acid, three changes of 70-90% ethanol since washing with water will make tissue swell and deteriorate

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Phloroglucin-Nitric acid

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Slides are brought to water and placed in 1% lithium carbonate for 1 hr and washed for 15 mins them stained

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HCl

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Also a softener. Produced good stain if 1% soltn with 70% alcohol for tissue blocks

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Von Ebner's

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Good cytologic staining

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Von Ebner's

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Dont require washing before dehy

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Von Ebner's fluid

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Recommended for teeth and small pieces of bones

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Von Ebner's fluid

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Decal cant be measured by chem test (same with perenyi's fluid)

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Formic acid

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Decal of postmortem research tissues

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Formic acid

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Addition of citrate with hasten decal process

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Formic acid

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Only used when there is minimal decalcification is required

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Formic acid

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Acts as fixative and decal

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Formic acid

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Excellent nuclear and cytoplasmic staining

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Formic acid

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Recommen for small pieces of bones and teeth. (Same with Von Ebner's fluid)

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Formic acid

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Hasten if the vol is inc to 25ml.

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Formic acid

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Req neutralization with 5%NaSO4

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Formic Acid-Sodium Citrate Soltn

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Recom for autopsy, BM, Cartilage, and tissue for research purp.

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Trichloroacetic acid

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Dont req washing out. Excess acid removed by several changes of 90% alcohol, which improves tx dehy

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Sulfuric acid

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Very weak decal only for minute pieces of bone

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Chromic acid / Flemming's fluid w/ HAC

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Fix and decal agent (same with formic acid)

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Chromic acid / Flemming's fluid w/ HAC

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Used for decal minute bone spicules

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Chromic acid / Flemming's fluid w/ HAC

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Nclr stn w/ H&E is inhibited

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Chromic acid / Flemming's fluid w/ HAC

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Tends to undergo reduction and forms ppt at bottom of container

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Chromic acid / Flemming's fluid w/ HAC

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Insol pigments are formed

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Chromic acid / Flemming's fluid w/ HAC

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Degree of decal cant be measured by routine chem test

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Citric acid-citrate buffer soltn ph 4.5

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Doesnt prod cell or tx distortion

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Citric acid-citrate buffer soltn ph 4.5

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Excellent nclr and cytopl stn

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EDTA

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Small tx is placed in it for 1-3wk

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EDTA

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Large tx is placed for 6-8wk

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Calcium will not bind

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Below pH3

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Faster binding of Ca

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pH 7-7.4

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Optimal Ca binding

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pH 8

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EDTA

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Excellent decal agnt for immunohistochemical enzyme stng and for EM

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EDTA

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Inact ALP

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To restore ALP act

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Add Magnesium Chloride

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IER

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Hastens decal of formic acid containing decal agnt

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IER

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Nitric or HCl containing decal agnt is not recom

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IER

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1/2 is spread over the bot of container. 20-30x vol of tx

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IER

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Tx allowed to stay in soltn for 11-14 days

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IER

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Decal degree measured by phy or x-ray mtd

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React Resin

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Immerse in N/10 HCl twice and wash w/ dist H20 3x

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IER

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Washing of soltn is eliminated

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IER

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Minimal histological artifacts (same w/ EDTA)

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Electrophoresis

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Satisfactory for small bone fragments

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At 37°C

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Impaired staining of Van Gieson's stn for collagen fibers

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At 55°C

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Undergo complete digestion w/in 24-48hrs

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18-30°C

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Optimal temp for decal

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24-48 hrs

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Ideal time for comp decal

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Lithium carbonate

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Used in post decal

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5-10% aqueous soltn. of Na bicarbonate

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Used in post decal(same w/ sat. lithium carbonate)

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Post decal with H2O

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30 mins for small tx

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Post decal with H2O

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1 to 4 hrs if large tx

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Acid decal for fzn sectn

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Formol saline containing 15% sucrose

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Acid decal for fzn sectn

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Or Phosphate-buffered saline with 15-20% sucrose

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Decal in EDTA

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Placed directly to alcohol

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Molliflex, 2% HCl, or 1% HCl in 70% alcohol

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Examples of tissue softeners