DECALCIFICATION Flashcards
procedure whereby CALCIUM or LIME SALTS are REMOVED from tissues that makes them flexible and easier to cut
DECALCIFICATION
RECOMMENDED temperature for decalcification
ROOM TEMP (18-30°C) [22-24°C]
IDEAL TIME required for decalcifying tissue
24-48 HOURS
VOLUME OF DECALCIFYING AGENT
20x
DECALCIFYING AGENTS: “ACIE”
- ACID
- CHELATING AGENTS
- ION-EXCHANGE RESIN
- ELECTRICAL IONIZATION / ELECTROPHORESIS
for routine purposes, what 2 ACIDS are used?
- NITRIC AICD
- FORMIC ACID
the MOST COMMON and FASTEST decalcifying agent used so far
NITRIC ACID
RECOMMENDED CONCENTRATION of Nitric Acid
5-10%
Concentrated solutions of Nitric Acid may inhibit nuclear stains & destroy tissues. To prevent this, what 2 reagents must be mixed with Nitric Acid?
FORMALDEHYDE or ALCOHOL
RECOMMENDED for URGENT BIOPSIES and for NEEDLE and SMALL BIOPSY SPX
10% AQUEOUS NITRIC ACID
aqueous sounds like “CUTE”? so small biopsy spx…
what decalcifying agent produces LESS TISSUE DISTORTION THAN 10% AQ. NITRIC ACID?
FORMOL NITRIC ACID
to remove the yellow color imparted by Formol Nitric Acid, neutralization with 5% of what substance is used?
5% SODIUM THIOSULFATE
in the neutralization to remove the yellow color imparted by Formol Nitric Acid, 5% sodium thiosulfate and what other substance is used?
RUNNING TAP WATER
this decalcifying agent is used for ROUTINE PURPOSES, NOT FOR URGENT PURPOSES
PERENYI’S FLUID
it DECALCIFIES and SOFTENS tissues at the same time
PERENYI’S FLUID
in Perenyi’s Fluid…
what is avoided due to the presence of CHROMIC ACID and ALCOHOL?
MACERATION
the MOST RAPID DECALCIFYING AGENT, recommended for URGENT WORKS
PHLOROGLUCIN NICTRIC ACID
what nitric acid decalcifying agents impart YELLOW COLOR to tissues?
- NITRIC ACID
- FORMOL NITRIC ACID
- PHLOROGLUCIN NITRIC ACID
decal agent inferior compared to nitric acid, but produces good nuclear staining
HYDROCHLORIC ACID
alternative for HYDROCHLORIC ACID
MURIATIC ACID
recommended for SURFACE DECALCIFICATION of tissue blocks if used in 1% solution with 70% alcohol
HYDROCHLORIC ACID
recommended for TEETH AND SMALL PIECES OF BONES
VON EBNER’S FLUID
composition of Von Ebner’s Fluid
- 30% SATURATED AQUEOUS NaCl
- CONCENTRATED HCl
- DISTILLED WATER
recommended for routine decalcification of POST-MORTEM RESEARCH TISSUES
FORMIC ACID
composition of AQUEOUS FORMIC ACID
FORMIC ACID + 10% FORMOL SALINE
Formic Acid recommended for small pieces of bones and teeth
not Von Ebner’s!
AQUEOUS FORMIC ACID
suitable for most ROUTINE SURGICAL SPECIMEN AND IMMUNOHISTOCHEMICAL STAINING
AQUEOUS FORMIC ACID
composition of FORMIC ACID-SODIUM CITRATE
20% AQUEOUS SODIUM CITRATE + 45% FORMIC ACID
recommended for AUTOPSY MATERIALS, BONE MARROW, CARTILAGE and TISSUES STUDIED FOR RESEARCH PURPOSES
FORMIC ACID-SODIUM CITRATE
decalcifying agent that DOES NOT NEED WASHING OUT
TRICHLOROACETIC ACID
WEAKEST DECALCIFYING AGENT; suitable only for minute pieces of bone
SULFUROUS ACID
used as a FIXATIVE & DECALCIFYING AGENT; decalcifies minute bone spicules
CHROMIC ACID
it is an environmental toxin, highly corrosive to skin and mucous membrane and carcinogenic
CHROMIC ACID
permits excellent nuclear and cytoplasmic staining
CITRIC ACID CITRATE BUFFER
buotan kaayo siya paminawon kay excellent ang nuclear & cytoplasmic
Citric Acid Citrate Buffer permits excellent nuclear and cytoplasmic staining at what pH?
pH 4.5
DOES NOT PRODUCE CELL OR TISSUE DISTORTION; too slow for routine purposes
CITRIC ACID CITRATE BUFFER
buotan kaayo siya paminawon kay di makadistort
substances which combine with calcium ions and other salts such as Fe and Mg to form weakly dissociated complexes and facilitate removal of calcium salt
CHELATING AGENTS
preferred for NUCLEIC ACID & NUCLEAR DNA PRESERVATION and ENZYME HISTOCHEMISTRY
CHELATING AGENTS
appropriate for IHC, FISH, and PCR where very high-quality morphology is required
CHELATING AGENTS
the MOST COMMON CHELATING AGENT
EDTA
COMMERCIAL NAME OF EDTA
VERSENE
chelating agent recommended ONLY FOR DETAILED MICROSCOPIC STUDIES
VERSENE
removes calcium ions from formic acid, thereby increasing solubility from the tissue
ION-EXCHANGE RESIN
NOT RECOMMENDED for fluids containing acids such as NITRIC ACID or HYDROCHLORIC ACID
ION-EXHANGE RESIN
process whereby positively charged Ca ions are attracted to a NEGATIVE ELECTRODE and subsequently removed from the decalcifying solution
ELECTRICAL IONIZATION (ELECTROPHORESIS)
Electrical Ionization is DEPENDENT on what current to remove calcium deposits?
DIRECT CURRENT
FASTER THAN ROUTINE DECALCIFICATION irrespective of the decalcifying agents used
MICROWAVE OVEN DECALCIFICATION
thickness of sections optimal for decalcification with or without microwave-accelerated processing
3-5 mm
what concentration of HCl is used for approximately 1hr to dissolve calcium in surface decalcification?
10% HCl
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
touching, bending, or squeezing the tissue with the fingers to determine the consistency of tissues
PHYSICAL / MECHANICAL
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
MOST IDEAL, MOST SENSITIVE, and MOST RELIABLE method
RADIOLOGICAL / X-RAY
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
method that DETECTS EVEN THE SMALLEST CALCIUM IONS
RADIOLOGICAL / X-RAY
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
EXCELLENT method for large specimens such as FEMORAL HEADS
RADIOLOGICAL / X-RAY
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
NOT RECOMMENDED FOR MERCURIC CHLORIDE-FIXED TISSUES
RADIOLOGICAL / X-RAY
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
SIMPLE, RELIABLE, and CONVENIENT METHOD RECOMMENDED FOR ROUTINE PURPOSES
CHEMICAL (CALCIUM OXALATE TEST)
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
another term for Chemical mtd
CALCIUM OXALATE TEST
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
what material is used as a pH indicator for the chemical method?
LITMUS PAPER
WAYS TO MEASURE THE EXTENT OF DECALCIFICATION:
what material is used as an ACID NEUTRALIZER in chemical method?
STRONG AMMONIA
in Chemical Method…
_____________ indicates INCOMPLETE DECALCIFICATION
CLOUDINESS
in Chemical Method…
what indicates a COMPLETE DECALCIFICATION?
CLEAR
5 TISSUE SOFTENERS
- PERENYI’S FLUID
- 4% PHENOL
- MOLLIFLEX
- 2% HCl
- 1% HCl in 70% ALCOHOL
3 solutions used in ELECTRICAL IONIZATION
- 88% FORMIC ACID
- CONCENTRATED HCl
- DISTILLED WATER