Decal

Ruffled border

What is the term for a delicate cytoplasmic tubules that are projections off of osteoclasts?

electrolytic

Microwave, sonication and electrolytic methods produce heat and must be carefully monitored to prevent excessive temperatures which damage tissue. [Bancroft]

Which decalcification method may cause burning of the specimen?

Water

-removes excess acid

After completion of decalcification, the specimen should be rinsed in

simple acid, electrolytic

Most bone specimens can be decalcified within 2 -6 hours, but possibly because of heat generated by the electrolytic method, there is a strong potential for tissue destruction and a total loss of cellular detail. [Carson]

With this method, there is a strong potential for tissue destruction and total loss of cellular detail and tissue stainability due to the heat generated.

bone marrow

The tissue shown here has cortical bone, is very dense and relatively acellular. (Carson, pg. 49, 128)

Remodeling and resorption

What are the two terms used in bone growth?

ion-exchange

Ion exchange resins involve the use of formic acid over a layer of an ammoniated salt of a sulfonated resin. Ammonium ions from the resin are exchanged for calcium ions; this keeps the solution free of calcium ions and speeds up the reaction. Staining is excellent after application of this method, and the time tissue remains in the decalcifying reagent is not critical. Because the solution remains relatively f free of calcium ions, it does not need to be changed frequently. This is one of the best, if nor the best, methods of decalcification. [Carson]

In which of the following methods does the decalcifying solution not have to be changed frequently?

acid

calcium oxide

chelating

ion-exchange

osteoclast

(a cell stained in red positive for Tartrate Resistant Acid Phosphatase.) This multinucleated cell is derived from monocytes and resorb bone. Which cell is this most likely to be?

False

High heat may cause tissue destruction and a loss of cellular detail, and therefore stain quality.

(T/F) Heating the solution to 60°C is a good method of increasing the rate of decalcification.

Keratin sulfate, Chondroitin sulfate, Hyaluronic acid

What is the protein core of the cartilage matrix?

processing schedule

Bone density, decal solution strength and solution temperature all influence the time necessary for decalcification to occur. Processing schedules have no impact on decalcification. [Carson ]

The amount of time a specimen needs to remain in the decalcifying solution does not depend upon which of the following?

articular

Which is not a bone type?

chemical, mechanical/physical, radiographic

According to Carson, list in alphabetical order the three methods to determine the decalcification has been completed

the overdecalcification seen in a section

The overdecalcification affected nuclear staining - nuclei are not as basophilic (blue) as they should be. (Carson, pg. 48-50)

Carefully checking the endpoint of decalcification would have prevented

chrondrocytes

Which is NOT a bone cell?

osteoblasts

osteoclasts

chrondrocytes

osteocytes

False

Zenker is a metallic fixative, and metals will render the specimen opaque so that the endpoint cannot be determined by X-ray. (Carson, pg. 48)

(T/F) The best method of checking Zenker-fixed tissue for the completeness of decalcification is radiologic examination.

Carson lists under "acid methods:" Simple acids, ion-exchange resins and electrolytic methods.

What are the three acid methods of decalcification?

Electrolytic

Which of the following methods is not used as a test for completion of decalcification?

Chemical

Electrolytic

Physical

Radiographic

surface decalcification

If an area of bone that is not completely decalcified is exposed during microtomy, satisfactory sections may be obtained by:

simple acid, electrolytic

Most bone specimens can be decalcified within 2 -6 hours, but possibly because of heat generated by the electrolytic method, there is a strong potential for tissue destruction and a total loss of cellular detail. [Carson]

With this method, there is a strong potential for tissue destruction and total loss of cellular detail and tissue stainability due to the heat generated.

False

Chelating agents bind ionized calcium ions, but no other ions are exchanged for the calcium ion. (Carson, pg. 48)

(T/F) A chelating agent exchanges another ion for the calcium ion.

nuclear basophilia

The most pronounced effect (of acid decalcification) is on nuclear basophilia. Overdecalcification may result in a total lack of nuclear staining; therefore, with many solutions the endpoint of decalcification must be carefully monitored. [Carson]

The most pronouced effect of over decalcification is on __________________

True

(T/F) Neither formic acid and nitric acid are used to check the endpoint of decalcification.

(1) fixation

(2) processing/dehydration

According to Carson, decalcification is accomplished after (1) and before (2)

False

Microtomy will be easier if the bone sections are embedded diagonally in the block. (Carson, pg.43)

(T/F) Bone sections should be embedded parallel to the long axis of the block.

most likely overdecalcification

What could be a problem of a section if it results in a lack of nuclear staining? (Carson, pg. 49-50)

True

Both acids may be used, although the process must be carefully monitored if using nitric acid. (Carson, pg.47)

(T/F) Both formic acid and nitric acid are used to decalcify specimens.

Nuclei

Rapid decalcifiers are more likely to adversely affect any subsequent staining, especially if not fixed completely. This is most noticeable in cell nuclei with the failure of nuclear chromatin to take up hematoxylin and other basic dyes as readily as soft tissues never exposed to acid solutions. The staining using acid dyes is also less affected, but eosin (an acid dye) can stain tissue a deep, unpleasant, brick red without the preferred three differential shades. [Bancroft]

The stronger the acidity of the solution and the longer the specimen remains in it, the more that subsequent staining will demonstrate injurious effects of the decalcification process. The most pronounced effect is on nuclear basophilia. [Carson]

The effect of acid decalcification is most noticeable in the staining of

trim the bone with a diamond blade saw

trim the bone surface after decalcification but before processing

To prevent or correct the bone dust artifact, on e must (more than one answer may be correct)

fixed

Carson: Best practice requires that bone specimens are cut into thin slices (approximately 4 mm) at the gross bench and well fixed before decalcification. Decalcification of specimens that are not well fixed may adversely affect morphology.

According to Carson, unless a decalcifying agent combined with a fixative is used, tissue must be thoroughly ______________ before it is submitted for decalcification

mesenchymal tissue

What type of tissue is hyaline and fibrocartilage developed from?

Ethylenediaminetetraacetic acid (EDTA)

Which of the following is a chelating agent used in decalcification?

bone

Overtime, what does most hyaline cartilage become?

False

Chelating agents bind ionized calcium ions, but no other ions are exchanged for the calcium ion. (Carson, pg. 48)

(T/F) A chelating agent exchanges another ion for the calcium ion.

radiography

Radiography is the most accurate method of determining the completeness of decalcification, and the small X-ray units available in many histology laboratories are easy to use. This method yields visual evidence that demineralization is complete. Radiographic methods cannot be used with metallic fixed tissue, such as zinc formalin, because the metal will render the specimen radiopaque.

What is the most accurate method to determine end-point of decalcification?

Endochrondral and intramembranous

What are two terms used to classify bone formation?

bone marrow

What lies between the spaces of spongy and compact bone?

Haversian canal

The osteon is a cylinder of bone, generally oriented parallel to the long axis of the bone. In the center of each osteon there is a canal called _________, which is lined by a endosteum - a single layer of bone cells that cover the bone surface; each canal houses blood vessels, nerves and loose CT.

4-5 mm

mostly 4 mm

For proper decalcification, bone should be cut at no more than ____________ in thickness.

binding ionized calcium ions/absorbing calcium ions, and must be changed frequently when used.

Acid decalcifiers act by forming soluble calcium salts, and chelating agents act by:

osteocytic lacuna

Osteocytes reside in lacunae and their processes in interconnecting canaliculi. [Tissue ID lecture notes]

Osteocytes are mature bone cells inside bone matrix. What are the spaces they occupy called?

osteocytic processes

Osteocytes connect with one another by way of their ____________________________, which occupy canaliculi.

Temperature, agitation, ratio of volume of decalcifying solution and specimen volume, specimen size, pH. Slide 39Sheehan states "The actual rate of decalcification varies with the size and density of the bone sample and can be hastened by occasional agitation or addition of fresh solution."

1) Thickness & Density

2) Temperature

3) Agitation

4) Solution strength or amount of solution

The time in solution does not change the speed of reaction (rate of reaction).

List FOUR factors influencing the rate of decalcification: